glp-1 Search Results


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  • 99
    Millipore glp 1
    Induction of glucose-dependent insulin secretion by (xGLP-E4). (A) INS-1 cells and (B) β-TC-6 cells were cultured in 24-well plates until 90% confluence. Cells were washed and incubated in Krebs-Ringer Bicarbonate (KRB) buffer and treated with glucose for 2 hours at 37℃. Cells were incubated with 10 nM glucagon-like peptide-1 <t>(GLP-1)</t> analog in KRB buffer containing low and high glucose for another 2 hours, and insulin secretion into the buffer was then determined. CTL, control; GLP-1, glucagon-like peptide-1; Exe-4, exendin-4; xGLP, Xenopus GLP-1.
    Glp 1, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1479 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Bachem glp 1
    <t>GLP-1</t> restores muscle metabolic insulin sensitivity during lipid infusion. Each rat received a systemic infusion of either saline or lipid for 240 min with a euglycemic insulin clamp (3 mU/kg/min) superimposed in the last 3 h and either saline or GLP-1 (30 pmol/kg/min) for the last 2 h. A : Time course of GIR ( n = 10–15/group). B : GIR AUC from 60–180 min; compared with insulin alone, * P
    Glp 1, supplied by Bachem, used in various techniques. Bioz Stars score: 92/100, based on 821 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Mimetics glp 1 mimetics
    Effects of MW1219 on insulin and <t>GLP-1</t> secretion in vitro . The effect of MW1219 to stimulate insulin secretion in MIN6 cells was studied in 2.8 mM and 16.8 mM glucose (A). Influence of GPR119 knockdown on MW1219-induced insulin secretion (C). Effect of MW1219 on GLP-1 release in GLUTag cells (B). After siRNA treatment, the effect of MW1219 on GLP-1 release was determined with GLUTag cells (D). All data points were representatives of three independent experiments, determined in triplicate. *P
    Glp 1 Mimetics, supplied by Mimetics, used in various techniques. Bioz Stars score: 93/100, based on 1082 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Mimetics glp 1 receptor agonists
    Placebo‐controlled clinical trials with <t>GLP‐1</t> receptor agonists exenatide, liraglutide, exenatide long‐acting release (LAR), AVE 0010 (lixisenatide) and taspoglutide on a background of metformin treatment in patients no longer controlled with a single oral antidiabetic drug. Effects on HbA 1c and fasting plasma glucose, the proportion of patients reaching a HbA 1c
    Glp 1 Receptor Agonists, supplied by Mimetics, used in various techniques. Bioz Stars score: 91/100, based on 158 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    glp 1  (Abcam)
    99
    Abcam glp 1
    Effects of sitagliptin (SG), exendin-3 (Ex-3), and liraglutide (Li) on the renal glomerulus structure and protein expression of dipeptidyl peptidase-4 (DPP-4) and glucagon-like peptide-1 <t>(GLP-1)</t> in the kidney during monocrotaline- (MCT-) induced renal injury . (a) Representative renal histological staining with haematoxylin and eosin (HE) and periodic acid-Schiff (PAS). (b) Graphic analysis of the average glomerulus surface area according to the HE staining. (c) Graphic analysis of the degree of glomerular mesangial expansion according to the PAS staining. (d) Representative western blots of DPP-4 and GLP-1. (e, f) Immunoblot analysis of DPP-4 and GLP-1. Data are expressed as the means ± SD; n = 6–8 rats in each group; # P
    Glp 1, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 201 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    glp 1  (LINCO)
    92
    LINCO glp 1
    Protocol design. The protocol consisted of a tracer equilibration period (0–90 min) and a basal period (90–120 min) followed by three test periods of 90 min each (P1 = 120–210 min, P2 = 210–300 min, and P3 = 300–390 min). The control group received saline, the PePe group received glucagon-like peptide 1 <t>(GLP-1;</t> 1 pmol·kg −1 ·min −1 ) peripherally, the PePo group received GLP-1 (1 pmol·kg −1 ·min −1 ) peripherally (P2) and then intraportally (P3), and the PeHa group received GLP-1 (1 pmol·kg −1 ·min −1 ) peripherally (P2) and then through the hepatic artery (P3) to increase the hepatic GLP-1 load to the same extent as in P3 in the PePo group ( n = 8 dogs/group). The GLP-1 infusion rate was 1 pmol·kg −1 ·min −1 in the PePe, PePo, and PeHa groups during P2 and P3.
    Glp 1, supplied by LINCO, used in various techniques. Bioz Stars score: 92/100, based on 386 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Prospec glp 1
    GLP-1R-dependent pathway and <t>GLP-1(9–36)-related</t> pathway are involved in effects of GLP-1 on eNOS levels in HUVECs. (A) GLP-1R and DPP-4 proteins were detected in HUVECs by Western blot analysis. Cells were incubated in the presence or absence (control) of GLP-1, GLP-1(9–36) or exenatide (GLP-1R agonist) (all at 5000 pmol/L) for the following times (B, C, D, E). Cells were incubated with GLP-1 in the presence of exendin (9–39) (G+E) or sitaglipin (G+S) or both (G+E+S) for the following times (F, G, H). (B, F) After 30-min incubation, eNOS activity was determined by NO content in cells. (C, G) After 5-min incubation, phosphorylation of eNOS at ser-1177 was examined by Western blot analysis. (D, H) After 48-h incubation, total eNOS protein level was measured by Western blot analysis. The upper parts of C, D, G, and H show representative experiments. Data are mean±SD after normalization to β-actin level. (E) After 48-h incubation, eNOS mRNA level was quantified by real-time RT-PCR. Data are mean±SD after normalization to β-actin level. b P
    Glp 1, supplied by Prospec, used in various techniques. Bioz Stars score: 92/100, based on 161 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Abcam glp 1r
    Expression and abundance of <t>GLP-1R</t> and GIPR in monocytes/macrophages and arterial SMCs. a Glp1r and ( b ) Gipr mRNA in exudate peritoneal macrophages and tissues as labelled from non-treated Apoe −/− mice were measured by real-time RT-PCR. c Western blotting analyses of GLP-1R and ( d ) GIPR abundance in exudate peritoneal macrophages of non-treated Apoe −/− mice and other mouse tissues as labelled. Three independent experiments were performed. e Immunostaining of GLP-1R or GIPR in exudate peritoneal macrophages from non-treated Apoe −/− mice. Green, GLP-1R or GIPR as indicated; red, nuclei; red + green, overlay of GLP-1R or GIPR and nuclei. Scale bars 50 μm. f GLP1R and ( g ) GIPR mRNA levels in human coronary artery SMCs, THP1 cells and THP1-derived macrophages. h GIPR mRNA level in human monocytes and monocyte-derived macrophages were measured by real-time RT-PCR. Results were obtained from three to six independent experiments
    Glp 1r, supplied by Abcam, used in various techniques. Bioz Stars score: 97/100, based on 375 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Phenomenex glp 1
    <t>GLP-1</t> formulated in PGC-C18 has extended blood circulation time compared to GLP-1 alone
    Glp 1, supplied by Phenomenex, used in various techniques. Bioz Stars score: 92/100, based on 164 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Meso Scale Diagnostics LLC glp 1
    ffar2 and ffar3 knockout mice have impaired glucose tolerance. A : Glucose stimulated <t>GLP-1</t> secretion in vivo. ffar2 −/− and ffar3 −/− mice and wild-type littermates ( n = 5 each) were dosed with DPP4 inhibitor at a dose of 20 mg/kg per os after a 4-h fast. Thirty minutes post–DPP4 inhibitor dosing, mice were dosed with 1.5 g/kg glucose per os Plasma active GLP-1 was assessed by a MesoScale assay at 0 and 30 min of the oral glucose tolerance test. Data represent means ± 1 SEM, and statistical significance was assessed by Student t test: * P
    Glp 1, supplied by Meso Scale Diagnostics LLC, used in various techniques. Bioz Stars score: 92/100, based on 192 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Phoenix Pharmaceuticals glp 1
    The mean activity of human gastric lipase (HGL) and mean concentrations of cholecystokinin (CCK), glucagon-like peptide-1 <t>(GLP-1)</t> and glucose-dependent insulinotropic peptide (GIP) in the study groups (non-HPG—non- Helicobacter pylori gastritis group, HPG— Helicobacter pylori gastritis group, CG—control group).
    Glp 1, supplied by Phoenix Pharmaceuticals, used in various techniques. Bioz Stars score: 94/100, based on 424 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    glp 1  (ALPCO)
    94
    ALPCO glp 1
    <t>GLP-1</t> response during OGTT per model. Data are shown as mean ± SEM. P values are for the comparison of the AUC for each group.
    Glp 1, supplied by ALPCO, used in various techniques. Bioz Stars score: 94/100, based on 340 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Drucker Diagnostics glp 1
    <t>GLP-1</t> actions and target organs . GLP-1, through its receptor GLP-1R, has functional effects on a variety of tissues.
    Glp 1, supplied by Drucker Diagnostics, used in various techniques. Bioz Stars score: 93/100, based on 256 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    21st Century Biochemicals glp 1
    A and B, Effects of vildagliptin and <t>GLP-1</t> given separately, GLP-1 in combination with vildagliptin, Ex-4, or saline on food intake in WT mice (A) and rats (B). Data are mean ± sem . *, P
    Glp 1, supplied by 21st Century Biochemicals, used in various techniques. Bioz Stars score: 92/100, based on 85 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Shibayagi glp 1
    Summary of single-fiber responses. A–E ) Response profiles of S-type <t>(GLP-1-sensitive:</t> n = 7, GLP-1-insensitive: n = 10), S-type after Exendin 4(3-39) treatment ( n = 9), electrolytes-responsive type (E-type, n = 13), NaCl-specific type (N-type,
    Glp 1, supplied by Shibayagi, used in various techniques. Bioz Stars score: 92/100, based on 93 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Merck & Co glp 1
    Plasma proinsulin levels before and after bolus injections of 5 g arginine under both basal and approximately 230 mg/dl hyperglycemic clamp conditions. A, Results for the islet transplant group (n = 5) during <t>GLP-1</t> 7–36 amide (▪) or placebo (□) infusion. B, Results for the pancreas transplant group (n = 6) during GLP-1 7–36 amide (•) or placebo (○) infusion.
    Glp 1, supplied by Merck & Co, used in various techniques. Bioz Stars score: 92/100, based on 60 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Peninsula Laboratories glp 1
    <t>GLP-1</t> potentiates exocytosis in B-cells. Increases in cell capacitance (ΔC m , bottom trace) elicited by progressively longer (5, 10, 15, 30, 50, 100, 150, 250, 350, 450, and 850 ms) depolarizations from −70 to 0 mV (V, top trace) under (A) control conditions and (B) 4 min after the inclusion of 10 nM GLP-1 in the extracellular medium. The interval between two successive depolarizations was 15 s for pulses ≤50 ms and 30 s for longer pulses. For clarity, only responses to the 30-, 100-, 250-, and 450-ms depolarizations are shown. The recording was performed using the perforated patch configuration. (C) Relationship between pulse duration ( t ) and increase in cell capacitance (ΔC m ) under control conditions and in the presence of 10 nM GLP-1 as indicated. The curves were derived by fitting Eq. 3 to the data points for depolarizations ≤350 ms. The dotted lines represent linear fits to the values measured in response to the two longest depolarizations. Data are mean values ± SEM of seven paired experiments. *P
    Glp 1, supplied by Peninsula Laboratories, used in various techniques. Bioz Stars score: 92/100, based on 87 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    PolyPeptide Laboratories glp 1
    MVR presented as the mean plateau value of the acoustic intensity (AI) in vastus lateralis muscle at basal and after 5-, 10-, and 60-min infusion ( A ) of <t>GLP-1</t> in the locally infused leg, ( B ) infusion of GLP-1 with coinfusion of octreotide in the locally
    Glp 1, supplied by PolyPeptide Laboratories, used in various techniques. Bioz Stars score: 92/100, based on 61 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Tocris glp 1 9 36 amide
    GLP-1R-mediated Ca 2+  signaling in HEK-GLP-1R cells. HEK-GLP-1R cells or wild-type HEK-293 cells were grown in ELISA 8-well strips (96-well format) and used either with or without loading with the Ca 2+  indicator, fluo-4 as indicated.  A ) Fluo-4-loaded HEK-GLP-1R cells were challenged with a range of concentrations of GLP-1 7–36 amide and fluorescence recorded as an index of [Ca 2+ ] i . Fluorescence was calibrated to [Ca 2+ ] i  as described to enable determination of agonist potency. The pEC 50  for the peak of GLP-1 7–36 amide-mediated elevations of [Ca 2+ ] i  was 9.61±0.25 (n = 3).  B ) Fluo-4-loaded HEK-GLP-1R cells were challenged with either buffer, GLP-1 7–36 amide (10 nM) or GLP-1 9–36 amide (10 µM). Alternatively, cells were pretreated with thapsigargin (2 µM, 5 min) to deplete intracellular Ca 2+  stores before challenge with GLP-1 9–36 amide (10 µM). Responses to 30 µM GLP-1 9–36 amide were similar to that evoked by 10 µM (data not shown).  C ) Fluo-4-loaded wild-type HEK-293 cells (i.e. cells without expression of the GLP-1R) were challenged with compound 2 (100 µM) in the absence or presence of pretreatment with thapsigargin (2 µM, 5 min).  D ) Fluo-4-loaded HEK-GLP-1R cells were challenged with either vehicle control (1% DMSO) or compound 2 (100 µM) in the absence or presence of pretreatment with thapsigargin (2 µM, 5 min).  E ) Fluo-4-loaded HEK-GLP-1R cells were challenged with vehicle control (1% DMSO), a concentration of either compound 2 (10 µM) or GLP-1 9–36 amide (1 µM) established to have little effect on [Ca 2+ ] i  or alternatively, compound 2 (10 µM) and GLP-1 9–36 amide (1 µM) in combination. Additionally, cells were challenged with either compound 2 (10 µM) or compound 2 (10 µM) and GLP-1 9–36 amide (1 µM) in combination following pretreatment with thapsigargin (2 µM, 5 min). DMSO was present in all conditions. All data are representative of 3 independent experiments showing similar results.
    Glp 1 9 36 Amide, supplied by Tocris, used in various techniques. Bioz Stars score: 93/100, based on 54 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    glp 1  (4Gene)
    92
    4Gene glp 1
    Amino acid sequence of <t>GLP-1</t> and exendin-4. GLP-1 consists of two active circulating forms, GLP-1 (7–36) amide and GLP-1 (7–37).
    Glp 1, supplied by 4Gene, used in various techniques. Bioz Stars score: 92/100, based on 53 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    AnaSpec glp 1
    GLP-1R silencing abrogates <t>GLP-1-induced</t> signalling pathway activation. ARPE-19 cells were transfected with scrambled (scr) siRNA control or GLP-1R siRNA and treated with 10 nmol/L GLP-1 for 10 minutes. Cells were then lysed and tested with antibody anti-GLP-1R, anti-phPKB, or anti-phERK1/2. Membranes were stripped and reprobed, respectively, with anti- β actin, anti-PKB, or antiERK1/2-antibodies. (a) Representative western blot analyses of three different experiments are shown. (b) Quantification of densitometries of western blot bands of GLP-1R. Data were expressed as mean ± SE of fold induction relative to β -actin ( n = 3). *** P
    Glp 1, supplied by AnaSpec, used in various techniques. Bioz Stars score: 92/100, based on 87 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Induction of glucose-dependent insulin secretion by (xGLP-E4). (A) INS-1 cells and (B) β-TC-6 cells were cultured in 24-well plates until 90% confluence. Cells were washed and incubated in Krebs-Ringer Bicarbonate (KRB) buffer and treated with glucose for 2 hours at 37℃. Cells were incubated with 10 nM glucagon-like peptide-1 (GLP-1) analog in KRB buffer containing low and high glucose for another 2 hours, and insulin secretion into the buffer was then determined. CTL, control; GLP-1, glucagon-like peptide-1; Exe-4, exendin-4; xGLP, Xenopus GLP-1.

    Journal: Endocrinology and Metabolism

    Article Title: A Novel Long-Acting Glucagon-Like Peptide-1 Agonist with Improved Efficacy in Insulin Secretion and β-Cell Growth

    doi: 10.3803/EnM.2014.29.3.320

    Figure Lengend Snippet: Induction of glucose-dependent insulin secretion by (xGLP-E4). (A) INS-1 cells and (B) β-TC-6 cells were cultured in 24-well plates until 90% confluence. Cells were washed and incubated in Krebs-Ringer Bicarbonate (KRB) buffer and treated with glucose for 2 hours at 37℃. Cells were incubated with 10 nM glucagon-like peptide-1 (GLP-1) analog in KRB buffer containing low and high glucose for another 2 hours, and insulin secretion into the buffer was then determined. CTL, control; GLP-1, glucagon-like peptide-1; Exe-4, exendin-4; xGLP, Xenopus GLP-1.

    Article Snippet: Stability of GLP-1 analogs To evaluate the stability of GLP-1 and its analogs against DPP-IV activity, peptides were incubated at an initial concentration of 100 nM in DMEM containing 10% FBS or in 100% FBS at 37℃ for 0, 12, 24, 48, 72, and 96 hours in the presence or absence of 0.2 mM DPP-IV inhibitor diprotin A (Sigma-Aldrich, St Louis, MO, USA).

    Techniques: Cell Culture, Incubation, CTL Assay

    Stability of glucagon-like peptide-1 (GLP-1) analogs. The stability of GLP-1 and GLP-1 analogs against dipeptidyl peptidase-IV (DPP-IV) activity was evaluated by incubating the individual peptides in medium containing 10% fetal bovine serum (FBS) (A) or in 100% FBS (B). Peptides were incubated at an initial concentration of 100 nM in Dulbecco's Modified Eagle's medium containing 10% FBS or 100% FBS at 37℃ in the presence or absence of 0.2 mM of the peptidase inhibitor diprotin A. Peptide activity was then assessed by measuring luciferase activity in cells expressing GLP1R and CRE-luc. Results are presented as mean±standard error of the mean of at least three independent experiments.

    Journal: Endocrinology and Metabolism

    Article Title: A Novel Long-Acting Glucagon-Like Peptide-1 Agonist with Improved Efficacy in Insulin Secretion and β-Cell Growth

    doi: 10.3803/EnM.2014.29.3.320

    Figure Lengend Snippet: Stability of glucagon-like peptide-1 (GLP-1) analogs. The stability of GLP-1 and GLP-1 analogs against dipeptidyl peptidase-IV (DPP-IV) activity was evaluated by incubating the individual peptides in medium containing 10% fetal bovine serum (FBS) (A) or in 100% FBS (B). Peptides were incubated at an initial concentration of 100 nM in Dulbecco's Modified Eagle's medium containing 10% FBS or 100% FBS at 37℃ in the presence or absence of 0.2 mM of the peptidase inhibitor diprotin A. Peptide activity was then assessed by measuring luciferase activity in cells expressing GLP1R and CRE-luc. Results are presented as mean±standard error of the mean of at least three independent experiments.

    Article Snippet: Stability of GLP-1 analogs To evaluate the stability of GLP-1 and its analogs against DPP-IV activity, peptides were incubated at an initial concentration of 100 nM in DMEM containing 10% FBS or in 100% FBS at 37℃ for 0, 12, 24, 48, 72, and 96 hours in the presence or absence of 0.2 mM DPP-IV inhibitor diprotin A (Sigma-Aldrich, St Louis, MO, USA).

    Techniques: Activity Assay, Incubation, Concentration Assay, Modification, Luciferase, Expressing

    Potency of glucagon-like peptide-1 (GLP-1) analogs toward GLP-1 receptor (GLP1R). Ligand potencies of GLP-1 analogs were examined using HEK293T cells expressing GLP1R. Cells were treated with increasing concentrations of GLP-1 analogs for 6 hours, and luciferase activity was measured. The data on the sigmoidal curves and EC 50 values are presented as means±standard error of the mean of at least three independent experiments. CRE-luc, cAMP response element-luciferase; xGLP, Xenopus GLP-1.

    Journal: Endocrinology and Metabolism

    Article Title: A Novel Long-Acting Glucagon-Like Peptide-1 Agonist with Improved Efficacy in Insulin Secretion and β-Cell Growth

    doi: 10.3803/EnM.2014.29.3.320

    Figure Lengend Snippet: Potency of glucagon-like peptide-1 (GLP-1) analogs toward GLP-1 receptor (GLP1R). Ligand potencies of GLP-1 analogs were examined using HEK293T cells expressing GLP1R. Cells were treated with increasing concentrations of GLP-1 analogs for 6 hours, and luciferase activity was measured. The data on the sigmoidal curves and EC 50 values are presented as means±standard error of the mean of at least three independent experiments. CRE-luc, cAMP response element-luciferase; xGLP, Xenopus GLP-1.

    Article Snippet: Stability of GLP-1 analogs To evaluate the stability of GLP-1 and its analogs against DPP-IV activity, peptides were incubated at an initial concentration of 100 nM in DMEM containing 10% FBS or in 100% FBS at 37℃ for 0, 12, 24, 48, 72, and 96 hours in the presence or absence of 0.2 mM DPP-IV inhibitor diprotin A (Sigma-Aldrich, St Louis, MO, USA).

    Techniques: Expressing, Luciferase, Activity Assay

    Induction of β-cell growth by (xGLP-E4). INS-1 cells were seeded in 12-well plates at a density of 4×10 4 cells/well. Every 2 days, the respective peptides were added to the cultures in fresh medium containing the appropriate concentration of glucose. (A) Six and (B) 10 days after cell seeding, cells were washed, harvested, and counted. Results are presented as mean±standard error of the mean of at least three independent experiments. GLP-1, glucagon-like peptide-1; Exe-4, exendin-4; xGLP, Xenopus GLP-1. a vs. control (CTL) ( P

    Journal: Endocrinology and Metabolism

    Article Title: A Novel Long-Acting Glucagon-Like Peptide-1 Agonist with Improved Efficacy in Insulin Secretion and β-Cell Growth

    doi: 10.3803/EnM.2014.29.3.320

    Figure Lengend Snippet: Induction of β-cell growth by (xGLP-E4). INS-1 cells were seeded in 12-well plates at a density of 4×10 4 cells/well. Every 2 days, the respective peptides were added to the cultures in fresh medium containing the appropriate concentration of glucose. (A) Six and (B) 10 days after cell seeding, cells were washed, harvested, and counted. Results are presented as mean±standard error of the mean of at least three independent experiments. GLP-1, glucagon-like peptide-1; Exe-4, exendin-4; xGLP, Xenopus GLP-1. a vs. control (CTL) ( P

    Article Snippet: Stability of GLP-1 analogs To evaluate the stability of GLP-1 and its analogs against DPP-IV activity, peptides were incubated at an initial concentration of 100 nM in DMEM containing 10% FBS or in 100% FBS at 37℃ for 0, 12, 24, 48, 72, and 96 hours in the presence or absence of 0.2 mM DPP-IV inhibitor diprotin A (Sigma-Aldrich, St Louis, MO, USA).

    Techniques: Concentration Assay, CTL Assay

    Plasma glucose and hormones. Mean ± SEM concentrations of plasma glucose ( A ), serum insulin ( B ), serum C-peptide ( C ), plasma total GLP-1 ( D ), plasma total ghrelin ( E ), and serum leptin ( F ) assessed before (averaged across the 0915- and 0930-h baseline values) and after intranasal administration (vertical dotted line) of oxytocin (24 IU; ● and solid lines) and placebo (vehicle; ○ and dotted lines). Subjects ate from a test breakfast from 1000 to 1030 h and ingested snacks under the pretext of a taste test from 1240 to 1250 h. Mean baseline values of both conditions are averaged to a common baseline ( n = 20). * P

    Journal: Diabetes

    Article Title: Oxytocin Reduces Reward-Driven Food Intake in Humans

    doi: 10.2337/db13-0663

    Figure Lengend Snippet: Plasma glucose and hormones. Mean ± SEM concentrations of plasma glucose ( A ), serum insulin ( B ), serum C-peptide ( C ), plasma total GLP-1 ( D ), plasma total ghrelin ( E ), and serum leptin ( F ) assessed before (averaged across the 0915- and 0930-h baseline values) and after intranasal administration (vertical dotted line) of oxytocin (24 IU; ● and solid lines) and placebo (vehicle; ○ and dotted lines). Subjects ate from a test breakfast from 1000 to 1030 h and ingested snacks under the pretext of a taste test from 1240 to 1250 h. Mean baseline values of both conditions are averaged to a common baseline ( n = 20). * P

    Article Snippet: Circulating concentrations of growth hormone and active GLP-1 (i.e., the intact form of GLP-1) were likewise comparable between conditions (all P > 0.14).

    Techniques:

    Changes (mean +SEM) in glucose, insulin, glucagon-like peptide (GLP)-1 and ghrelin from the baseline values after administration of placebo (broken lines and open markings) or whey protein (solid lines and filled markings) during acute challenge tests.

    Journal: BMJ Open Diabetes Research & Care

    Article Title: Glucose-lowering effect of whey protein depends upon clinical characteristics of patients with type 2 diabetes

    doi: 10.1136/bmjdrc-2017-000420

    Figure Lengend Snippet: Changes (mean +SEM) in glucose, insulin, glucagon-like peptide (GLP)-1 and ghrelin from the baseline values after administration of placebo (broken lines and open markings) or whey protein (solid lines and filled markings) during acute challenge tests.

    Article Snippet: Total GLP-1 and ghrelin were measured by ELISA (Millipore, Billerica, Massachusetts, USA) with CVs of 3% and 2%, respectively.

    Techniques:

    GLP-1 restores muscle metabolic insulin sensitivity during lipid infusion. Each rat received a systemic infusion of either saline or lipid for 240 min with a euglycemic insulin clamp (3 mU/kg/min) superimposed in the last 3 h and either saline or GLP-1 (30 pmol/kg/min) for the last 2 h. A : Time course of GIR ( n = 10–15/group). B : GIR AUC from 60–180 min; compared with insulin alone, * P

    Journal: Diabetes

    Article Title: Glucagon-Like Peptide 1 Recruits Muscle Microvasculature and Improves Insulin’s Metabolic Action in the Presence of Insulin Resistance

    doi: 10.2337/db13-1597

    Figure Lengend Snippet: GLP-1 restores muscle metabolic insulin sensitivity during lipid infusion. Each rat received a systemic infusion of either saline or lipid for 240 min with a euglycemic insulin clamp (3 mU/kg/min) superimposed in the last 3 h and either saline or GLP-1 (30 pmol/kg/min) for the last 2 h. A : Time course of GIR ( n = 10–15/group). B : GIR AUC from 60–180 min; compared with insulin alone, * P

    Article Snippet: At the rates selected, insulin and GLP-1 each potently recruit muscle microvasculature ( , , , ), and Intralipid + heparin abrogates insulin-mediated muscle microvascular and metabolic responses ( , ).

    Techniques:

    GLP-1 infusion increases muscle insulin delivery and improves muscle metabolic response to insulin in HFD-fed rats. Each rat was fed either an LFD or HFD for 4 weeks. A : Time course of GIR. B : GIR AUC (60–180 min); n = 8–10/group. C : Muscle 125 I-insulin uptake; n = 6–11/group. D : Muscle Akt phosphorylation. E : Muscle eNOS phosphorylation. F : Muscle ERK1/2 phosphorylation. G : Aorta PKA phosphorylation. n = 6–8/group for D – G . Compared with LFD + Insulin, * P

    Journal: Diabetes

    Article Title: Glucagon-Like Peptide 1 Recruits Muscle Microvasculature and Improves Insulin’s Metabolic Action in the Presence of Insulin Resistance

    doi: 10.2337/db13-1597

    Figure Lengend Snippet: GLP-1 infusion increases muscle insulin delivery and improves muscle metabolic response to insulin in HFD-fed rats. Each rat was fed either an LFD or HFD for 4 weeks. A : Time course of GIR. B : GIR AUC (60–180 min); n = 8–10/group. C : Muscle 125 I-insulin uptake; n = 6–11/group. D : Muscle Akt phosphorylation. E : Muscle eNOS phosphorylation. F : Muscle ERK1/2 phosphorylation. G : Aorta PKA phosphorylation. n = 6–8/group for D – G . Compared with LFD + Insulin, * P

    Article Snippet: At the rates selected, insulin and GLP-1 each potently recruit muscle microvasculature ( , , , ), and Intralipid + heparin abrogates insulin-mediated muscle microvascular and metabolic responses ( , ).

    Techniques:

    GLP-1 increases muscle 125 I-insulin uptake. Each rat received a 90-min euglycemic insulin clamp (3 mU/kg/min) with or without simultaneous GLP-1 infusion (30 pmol/kg/min) for the last 30 min. A bolus intravenous injection of 125 I-insulin (1.5 µCi) was given 5 min before the end of insulin ± GLP-1 infusions. Blood and skeletal muscle were collected for determination of intact 125 I-insulin. A : Study protocol. B : Fraction of intact 125 I-insulin in blood and muscle. C : Muscle insulin uptake. n = 5 each. Compared with insulin alone, * P

    Journal: Diabetes

    Article Title: Glucagon-Like Peptide 1 Recruits Muscle Microvasculature and Improves Insulin’s Metabolic Action in the Presence of Insulin Resistance

    doi: 10.2337/db13-1597

    Figure Lengend Snippet: GLP-1 increases muscle 125 I-insulin uptake. Each rat received a 90-min euglycemic insulin clamp (3 mU/kg/min) with or without simultaneous GLP-1 infusion (30 pmol/kg/min) for the last 30 min. A bolus intravenous injection of 125 I-insulin (1.5 µCi) was given 5 min before the end of insulin ± GLP-1 infusions. Blood and skeletal muscle were collected for determination of intact 125 I-insulin. A : Study protocol. B : Fraction of intact 125 I-insulin in blood and muscle. C : Muscle insulin uptake. n = 5 each. Compared with insulin alone, * P

    Article Snippet: At the rates selected, insulin and GLP-1 each potently recruit muscle microvasculature ( , , , ), and Intralipid + heparin abrogates insulin-mediated muscle microvascular and metabolic responses ( , ).

    Techniques: Injection

    GLP-1 recruits muscle microvasculature in the presence of acute and chronic insulin resistance. A : Infusion protocol. Each rat received a 3-h infusion of GLP-1 (30 pmol/kg/min) or equal volume of saline. B – D : GLP-1–mediated changes in muscle microvascular parameters in the presence of acute insulin resistance induced by 4 h of lipid infusion. n = 4–5. E – G : GLP-1–mediated changes in muscle microvascular parameters in the presence of chronic insulin resistance induced by 4 weeks of HFD feeding. n = 4–7. Compared with 0 min, * P

    Journal: Diabetes

    Article Title: Glucagon-Like Peptide 1 Recruits Muscle Microvasculature and Improves Insulin’s Metabolic Action in the Presence of Insulin Resistance

    doi: 10.2337/db13-1597

    Figure Lengend Snippet: GLP-1 recruits muscle microvasculature in the presence of acute and chronic insulin resistance. A : Infusion protocol. Each rat received a 3-h infusion of GLP-1 (30 pmol/kg/min) or equal volume of saline. B – D : GLP-1–mediated changes in muscle microvascular parameters in the presence of acute insulin resistance induced by 4 h of lipid infusion. n = 4–5. E – G : GLP-1–mediated changes in muscle microvascular parameters in the presence of chronic insulin resistance induced by 4 weeks of HFD feeding. n = 4–7. Compared with 0 min, * P

    Article Snippet: At the rates selected, insulin and GLP-1 each potently recruit muscle microvasculature ( , , , ), and Intralipid + heparin abrogates insulin-mediated muscle microvascular and metabolic responses ( , ).

    Techniques:

    Lipid infusion abrogates insulin-mediated but not GLP-1 + insulin–mediated muscle microvascular perfusion. A : Infusion protocol. Each rat received a systemic infusion of either saline or Intralipid + heparin for 240 min with a euglycemic insulin clamp (3 mU/kg/min) superimposed in the last 3 h and either saline or GLP-1 (30 pmol/kg/min) for the last 2 h. B : MBV. C : MFV. D : MBF; n = 10 each. E : Muscle oxygen saturation. n = 5–6. F : Plasma NO levels. n = 4–8. Compared with respective baseline (0 min), * P

    Journal: Diabetes

    Article Title: Glucagon-Like Peptide 1 Recruits Muscle Microvasculature and Improves Insulin’s Metabolic Action in the Presence of Insulin Resistance

    doi: 10.2337/db13-1597

    Figure Lengend Snippet: Lipid infusion abrogates insulin-mediated but not GLP-1 + insulin–mediated muscle microvascular perfusion. A : Infusion protocol. Each rat received a systemic infusion of either saline or Intralipid + heparin for 240 min with a euglycemic insulin clamp (3 mU/kg/min) superimposed in the last 3 h and either saline or GLP-1 (30 pmol/kg/min) for the last 2 h. B : MBV. C : MFV. D : MBF; n = 10 each. E : Muscle oxygen saturation. n = 5–6. F : Plasma NO levels. n = 4–8. Compared with respective baseline (0 min), * P

    Article Snippet: At the rates selected, insulin and GLP-1 each potently recruit muscle microvasculature ( , , , ), and Intralipid + heparin abrogates insulin-mediated muscle microvascular and metabolic responses ( , ).

    Techniques:

    HFD feeding abolishes insulin-mediated but not GLP-1 + insulin–mediated muscle microvascular perfusion. Each rat was fed an LFD or HFD for 4 weeks. A : Infusion protocol. B : MBV. C : MFV. D : MBF; n = 9 to 10/group. E : Muscle oxygen saturation; n = 4–8/group. F : Plasma NO levels; n = 4–8/group. Compared with respective baseline (0 min), * P

    Journal: Diabetes

    Article Title: Glucagon-Like Peptide 1 Recruits Muscle Microvasculature and Improves Insulin’s Metabolic Action in the Presence of Insulin Resistance

    doi: 10.2337/db13-1597

    Figure Lengend Snippet: HFD feeding abolishes insulin-mediated but not GLP-1 + insulin–mediated muscle microvascular perfusion. Each rat was fed an LFD or HFD for 4 weeks. A : Infusion protocol. B : MBV. C : MFV. D : MBF; n = 9 to 10/group. E : Muscle oxygen saturation; n = 4–8/group. F : Plasma NO levels; n = 4–8/group. Compared with respective baseline (0 min), * P

    Article Snippet: At the rates selected, insulin and GLP-1 each potently recruit muscle microvasculature ( , , , ), and Intralipid + heparin abrogates insulin-mediated muscle microvascular and metabolic responses ( , ).

    Techniques:

    GLP-1 enhances insulin-mediated glucose disposal and muscle microvascular recruitment. Each rat received a 2-h euglycemic insulin clamp (3 mU/kg/min) for 120 min with or without GLP-1 infusion (30 pmol/kg/min) superimposed between 60 and 120 min. CEU measurements were done at 0, 60, 90, and 120 min. A : Study protocol. B : GIR during insulin clamp. C : GIR AUC between 60 and 120 min. D : MBV. E : MFV. F : MBF. G : Plasma NO levels. n = 4–15 each. Compared with 0 min, * P

    Journal: Diabetes

    Article Title: Glucagon-Like Peptide 1 Recruits Muscle Microvasculature and Improves Insulin’s Metabolic Action in the Presence of Insulin Resistance

    doi: 10.2337/db13-1597

    Figure Lengend Snippet: GLP-1 enhances insulin-mediated glucose disposal and muscle microvascular recruitment. Each rat received a 2-h euglycemic insulin clamp (3 mU/kg/min) for 120 min with or without GLP-1 infusion (30 pmol/kg/min) superimposed between 60 and 120 min. CEU measurements were done at 0, 60, 90, and 120 min. A : Study protocol. B : GIR during insulin clamp. C : GIR AUC between 60 and 120 min. D : MBV. E : MFV. F : MBF. G : Plasma NO levels. n = 4–15 each. Compared with 0 min, * P

    Article Snippet: At the rates selected, insulin and GLP-1 each potently recruit muscle microvasculature ( , , , ), and Intralipid + heparin abrogates insulin-mediated muscle microvascular and metabolic responses ( , ).

    Techniques:

    Effects of MW1219 on insulin and GLP-1 secretion in vitro . The effect of MW1219 to stimulate insulin secretion in MIN6 cells was studied in 2.8 mM and 16.8 mM glucose (A). Influence of GPR119 knockdown on MW1219-induced insulin secretion (C). Effect of MW1219 on GLP-1 release in GLUTag cells (B). After siRNA treatment, the effect of MW1219 on GLP-1 release was determined with GLUTag cells (D). All data points were representatives of three independent experiments, determined in triplicate. *P

    Journal: PLoS ONE

    Article Title: High-Throughput Screening for GPR119 Modulators Identifies a Novel Compound with Anti-Diabetic Efficacy in db/db Mice

    doi: 10.1371/journal.pone.0063861

    Figure Lengend Snippet: Effects of MW1219 on insulin and GLP-1 secretion in vitro . The effect of MW1219 to stimulate insulin secretion in MIN6 cells was studied in 2.8 mM and 16.8 mM glucose (A). Influence of GPR119 knockdown on MW1219-induced insulin secretion (C). Effect of MW1219 on GLP-1 release in GLUTag cells (B). After siRNA treatment, the effect of MW1219 on GLP-1 release was determined with GLUTag cells (D). All data points were representatives of three independent experiments, determined in triplicate. *P

    Article Snippet: Incretin-based therapies are also becoming popular which use either GLP-1 mimetics or DPP-4 inhibitors .

    Techniques: In Vitro

    Activities of MW1219 on insulin and GLP-1 secretion in db/db mice. Following 6 weeks of oral treatment with MW1219, (A) insulin and (B) GLP-1 levels after an oral glucose bonus were determined. *P

    Journal: PLoS ONE

    Article Title: High-Throughput Screening for GPR119 Modulators Identifies a Novel Compound with Anti-Diabetic Efficacy in db/db Mice

    doi: 10.1371/journal.pone.0063861

    Figure Lengend Snippet: Activities of MW1219 on insulin and GLP-1 secretion in db/db mice. Following 6 weeks of oral treatment with MW1219, (A) insulin and (B) GLP-1 levels after an oral glucose bonus were determined. *P

    Article Snippet: Incretin-based therapies are also becoming popular which use either GLP-1 mimetics or DPP-4 inhibitors .

    Techniques: Mouse Assay

    Amino acid sequence of GAlbudAb and activity of GAlbudAb. The sequence was aligned with that of the amino acid sequence for the human DPK9 - Jk1 germline gene, the GLP-1 (7–36) peptide and exendin-4 using the Clustal V algorithm. Residue numbering was determined by the method of Kabat (A) . Activity of native GLP-1 (7–36) and GAlbudAb in CHO reporter assay (B) .

    Journal: Cardiovascular Diabetology

    Article Title: Novel fusion of GLP-1 with a domain antibody to serum albumin prolongs protection against myocardial ischemia/reperfusion injury in the rat

    doi: 10.1186/1475-2840-12-148

    Figure Lengend Snippet: Amino acid sequence of GAlbudAb and activity of GAlbudAb. The sequence was aligned with that of the amino acid sequence for the human DPK9 - Jk1 germline gene, the GLP-1 (7–36) peptide and exendin-4 using the Clustal V algorithm. Residue numbering was determined by the method of Kabat (A) . Activity of native GLP-1 (7–36) and GAlbudAb in CHO reporter assay (B) .

    Article Snippet: GLP-1 regulates glucose homeostasis by stimulating insulin secretion, inhibiting glucagon secretion, delaying gastric emptying and promoting satiety [ ].

    Techniques: Sequencing, Activity Assay, Reporter Assay

    Placebo‐controlled clinical trials with GLP‐1 receptor agonists exenatide, liraglutide, exenatide long‐acting release (LAR), AVE 0010 (lixisenatide) and taspoglutide on a background of metformin treatment in patients no longer controlled with a single oral antidiabetic drug. Effects on HbA 1c and fasting plasma glucose, the proportion of patients reaching a HbA 1c

    Journal: Journal of Diabetes Investigation

    Article Title: Comparative evaluation of incretin-based antidiabetic medications and alternative therapies to be added to metformin in the case of monotherapy failure †

    doi: 10.1111/j.2040-1124.2010.00004.x

    Figure Lengend Snippet: Placebo‐controlled clinical trials with GLP‐1 receptor agonists exenatide, liraglutide, exenatide long‐acting release (LAR), AVE 0010 (lixisenatide) and taspoglutide on a background of metformin treatment in patients no longer controlled with a single oral antidiabetic drug. Effects on HbA 1c and fasting plasma glucose, the proportion of patients reaching a HbA 1c

    Article Snippet: Incretin‐Based Antidiabetic Medications Two novel classes of antidiabetic medications make use of the antidiabetic properties of the incretin hormone glucagon‐like peptide‐1 (GLP‐1) , the GLP‐1 receptor agonists (or incretin mimetics) and inhibitors of the protease dipeptidyl peptidase‐4 (DPP‐4; ‘incretin enhancers’) .

    Techniques:

    Amino acid sequence of glucagon‐like peptide‐1 and peptide GLP‐1 receptor agonists exenatide, liraglutide, and taspoglutide and chemical structures of the dipeptidyl peptidase‐4 inhibitors sitagliptin, vildagliptin, saxagliptin, and alogliptin.

    Journal: Journal of Diabetes Investigation

    Article Title: Comparative evaluation of incretin-based antidiabetic medications and alternative therapies to be added to metformin in the case of monotherapy failure †

    doi: 10.1111/j.2040-1124.2010.00004.x

    Figure Lengend Snippet: Amino acid sequence of glucagon‐like peptide‐1 and peptide GLP‐1 receptor agonists exenatide, liraglutide, and taspoglutide and chemical structures of the dipeptidyl peptidase‐4 inhibitors sitagliptin, vildagliptin, saxagliptin, and alogliptin.

    Article Snippet: Incretin‐Based Antidiabetic Medications Two novel classes of antidiabetic medications make use of the antidiabetic properties of the incretin hormone glucagon‐like peptide‐1 (GLP‐1) , the GLP‐1 receptor agonists (or incretin mimetics) and inhibitors of the protease dipeptidyl peptidase‐4 (DPP‐4; ‘incretin enhancers’) .

    Techniques: Sequencing

    Effects of sitagliptin (SG), exendin-3 (Ex-3), and liraglutide (Li) on the renal glomerulus structure and protein expression of dipeptidyl peptidase-4 (DPP-4) and glucagon-like peptide-1 (GLP-1) in the kidney during monocrotaline- (MCT-) induced renal injury . (a) Representative renal histological staining with haematoxylin and eosin (HE) and periodic acid-Schiff (PAS). (b) Graphic analysis of the average glomerulus surface area according to the HE staining. (c) Graphic analysis of the degree of glomerular mesangial expansion according to the PAS staining. (d) Representative western blots of DPP-4 and GLP-1. (e, f) Immunoblot analysis of DPP-4 and GLP-1. Data are expressed as the means ± SD; n = 6–8 rats in each group; # P

    Journal: BioMed Research International

    Article Title: Glucagon-Like Peptide-1 Mediates the Protective Effect of the Dipeptidyl Peptidase IV Inhibitor on Renal Fibrosis via Reducing the Phenotypic Conversion of Renal Microvascular Cells in Monocrotaline-Treated Rats

    doi: 10.1155/2018/1864107

    Figure Lengend Snippet: Effects of sitagliptin (SG), exendin-3 (Ex-3), and liraglutide (Li) on the renal glomerulus structure and protein expression of dipeptidyl peptidase-4 (DPP-4) and glucagon-like peptide-1 (GLP-1) in the kidney during monocrotaline- (MCT-) induced renal injury . (a) Representative renal histological staining with haematoxylin and eosin (HE) and periodic acid-Schiff (PAS). (b) Graphic analysis of the average glomerulus surface area according to the HE staining. (c) Graphic analysis of the degree of glomerular mesangial expansion according to the PAS staining. (d) Representative western blots of DPP-4 and GLP-1. (e, f) Immunoblot analysis of DPP-4 and GLP-1. Data are expressed as the means ± SD; n = 6–8 rats in each group; # P

    Article Snippet: After that, the PVDF membranes were hybridized in 5% nonfat dry milk or 5% bovine serum albumin (BSA) for 1 h at room temperature and were incubated overnight at 4°C with antibodies against DPP-4 (1 : 1000, Abcam PLC, Cambridge, MA, USA), GLP-1 (1 : 1000, Abcam PLC), cleaved-caspase 3 (1 : 1000; Cell Signaling Technology, Danvers, MA, USA), glucose-regulated protein 78 (GRP78, 1 : 1000, ProteinTech Group, Inc., Chicago, IL, USA), Bax (1 : 1000; Cell Signaling Technology), Bcl2 (1 : 1000; Cell Signaling Technology), smooth muscle 22 alpha (SM22α , 1 : 1000, ProteinTech Group), von Willebrand factor (vWF, 1 : 1000, ProteinTech Group), α -smooth muscle actin (α SMA, 1 : 1000, Abcam PLC), transforming growth factor-β 1 (TGF-β 1, 1 : 1000; Cell Signaling Technology), TGFβ receptor 1 (TGFβ R1, 1 : 1000, Abcam PLC), Smad3 (1 : 1000; Cell Signaling Technology); phosphorylated Smad3 (1 : 1000; Cell Signaling Technology), Snail (1 : 1000; Cell Signaling Technology), bone morphogenetic protein receptor type 2 (BMPR2, 1 : 1000, Abcam PLC), and β -actin (1 : 5000, ProteinTech Group).

    Techniques: Expressing, Staining, Western Blot

    Dipeptidyl peptidase-4 (DPP-4) inhibition with sitagliptin decreases the degranulation of glucagon-like peptide-1 (GLP-1) 7-36 (active form) and promotes the transduction of GLP-1R signalling, which can be blocked by the GLP-1R antagonist exendin-3. The GLP-1R agonist liraglutide can activate GLP-1R signalling directly. In smooth muscle cells (SMCs), the activation of GLP-1R upregulates the expression of smooth muscle 22 alpha (SM22 α ), which is expressed at high levels in the contractile phenotype of SMCs, and inhibits the transition of SMCs from the contractile phenotype to the synthetic phenotype. The extracellular matrix derived from the synthetic phenotype of SMCs is then obviously reduced. In endothelial cells, activated GLP-1R signalling can upregulate bone morphogenetic protein receptor type 2 (BMPR2) expression and reduce transforming growth factor- β 1 (TGF- β 1)/Smad3 signalling, followed by inhibiting Snail expression. Then, the protein expression of endothelial marker von Willebrand factor (vWF) increases, while that of the mesenchymal marker α -smooth muscle actin ( α SMA) decreases. After that, the endothelial-mesenchymal transition (EndMT) programme is blocked, the extracellular matrix derived from myofibroblasts is reduced, and, ultimately, renal fibrosis is attenuated.

    Journal: BioMed Research International

    Article Title: Glucagon-Like Peptide-1 Mediates the Protective Effect of the Dipeptidyl Peptidase IV Inhibitor on Renal Fibrosis via Reducing the Phenotypic Conversion of Renal Microvascular Cells in Monocrotaline-Treated Rats

    doi: 10.1155/2018/1864107

    Figure Lengend Snippet: Dipeptidyl peptidase-4 (DPP-4) inhibition with sitagliptin decreases the degranulation of glucagon-like peptide-1 (GLP-1) 7-36 (active form) and promotes the transduction of GLP-1R signalling, which can be blocked by the GLP-1R antagonist exendin-3. The GLP-1R agonist liraglutide can activate GLP-1R signalling directly. In smooth muscle cells (SMCs), the activation of GLP-1R upregulates the expression of smooth muscle 22 alpha (SM22 α ), which is expressed at high levels in the contractile phenotype of SMCs, and inhibits the transition of SMCs from the contractile phenotype to the synthetic phenotype. The extracellular matrix derived from the synthetic phenotype of SMCs is then obviously reduced. In endothelial cells, activated GLP-1R signalling can upregulate bone morphogenetic protein receptor type 2 (BMPR2) expression and reduce transforming growth factor- β 1 (TGF- β 1)/Smad3 signalling, followed by inhibiting Snail expression. Then, the protein expression of endothelial marker von Willebrand factor (vWF) increases, while that of the mesenchymal marker α -smooth muscle actin ( α SMA) decreases. After that, the endothelial-mesenchymal transition (EndMT) programme is blocked, the extracellular matrix derived from myofibroblasts is reduced, and, ultimately, renal fibrosis is attenuated.

    Article Snippet: After that, the PVDF membranes were hybridized in 5% nonfat dry milk or 5% bovine serum albumin (BSA) for 1 h at room temperature and were incubated overnight at 4°C with antibodies against DPP-4 (1 : 1000, Abcam PLC, Cambridge, MA, USA), GLP-1 (1 : 1000, Abcam PLC), cleaved-caspase 3 (1 : 1000; Cell Signaling Technology, Danvers, MA, USA), glucose-regulated protein 78 (GRP78, 1 : 1000, ProteinTech Group, Inc., Chicago, IL, USA), Bax (1 : 1000; Cell Signaling Technology), Bcl2 (1 : 1000; Cell Signaling Technology), smooth muscle 22 alpha (SM22α , 1 : 1000, ProteinTech Group), von Willebrand factor (vWF, 1 : 1000, ProteinTech Group), α -smooth muscle actin (α SMA, 1 : 1000, Abcam PLC), transforming growth factor-β 1 (TGF-β 1, 1 : 1000; Cell Signaling Technology), TGFβ receptor 1 (TGFβ R1, 1 : 1000, Abcam PLC), Smad3 (1 : 1000; Cell Signaling Technology); phosphorylated Smad3 (1 : 1000; Cell Signaling Technology), Snail (1 : 1000; Cell Signaling Technology), bone morphogenetic protein receptor type 2 (BMPR2, 1 : 1000, Abcam PLC), and β -actin (1 : 5000, ProteinTech Group).

    Techniques: Inhibition, Transduction, Activation Assay, Expressing, Derivative Assay, Marker

    Protocol design. The protocol consisted of a tracer equilibration period (0–90 min) and a basal period (90–120 min) followed by three test periods of 90 min each (P1 = 120–210 min, P2 = 210–300 min, and P3 = 300–390 min). The control group received saline, the PePe group received glucagon-like peptide 1 (GLP-1; 1 pmol·kg −1 ·min −1 ) peripherally, the PePo group received GLP-1 (1 pmol·kg −1 ·min −1 ) peripherally (P2) and then intraportally (P3), and the PeHa group received GLP-1 (1 pmol·kg −1 ·min −1 ) peripherally (P2) and then through the hepatic artery (P3) to increase the hepatic GLP-1 load to the same extent as in P3 in the PePo group ( n = 8 dogs/group). The GLP-1 infusion rate was 1 pmol·kg −1 ·min −1 in the PePe, PePo, and PeHa groups during P2 and P3.

    Journal: American journal of physiology. Gastrointestinal and liver physiology

    Article Title: Insulin secretion-independent effects of GLP-1 on canine liver glucose metabolism do not involve portal vein GLP-1 receptors

    doi: 10.1152/ajpgi.00121.2005

    Figure Lengend Snippet: Protocol design. The protocol consisted of a tracer equilibration period (0–90 min) and a basal period (90–120 min) followed by three test periods of 90 min each (P1 = 120–210 min, P2 = 210–300 min, and P3 = 300–390 min). The control group received saline, the PePe group received glucagon-like peptide 1 (GLP-1; 1 pmol·kg −1 ·min −1 ) peripherally, the PePo group received GLP-1 (1 pmol·kg −1 ·min −1 ) peripherally (P2) and then intraportally (P3), and the PeHa group received GLP-1 (1 pmol·kg −1 ·min −1 ) peripherally (P2) and then through the hepatic artery (P3) to increase the hepatic GLP-1 load to the same extent as in P3 in the PePo group ( n = 8 dogs/group). The GLP-1 infusion rate was 1 pmol·kg −1 ·min −1 in the PePe, PePo, and PeHa groups during P2 and P3.

    Article Snippet: Differences (P2 − P1 and P3 − P1) were calculated to assess the specific effect of GLP-1 over the effect achieved by pancreatic hormones and hyperglycemia per se.

    Techniques:

    Arterial plasma glucose and hepatic glucose loads in 42-h fasted dogs in the presence of somatostatin, intraportal insulin and glucagon, and peripheral glucose infusions. GLP-1 was infused peripherally during P2 and P3 (PePe group), peripherally during P2 and intraportally during P3 (PePo group), or peripherally during P2 and in the hepatic artery during P3 (PeHa group). Data are expressed as means ± SE; n = 8 dogs/group. There were no significant differences among the groups.

    Journal: American journal of physiology. Gastrointestinal and liver physiology

    Article Title: Insulin secretion-independent effects of GLP-1 on canine liver glucose metabolism do not involve portal vein GLP-1 receptors

    doi: 10.1152/ajpgi.00121.2005

    Figure Lengend Snippet: Arterial plasma glucose and hepatic glucose loads in 42-h fasted dogs in the presence of somatostatin, intraportal insulin and glucagon, and peripheral glucose infusions. GLP-1 was infused peripherally during P2 and P3 (PePe group), peripherally during P2 and intraportally during P3 (PePo group), or peripherally during P2 and in the hepatic artery during P3 (PeHa group). Data are expressed as means ± SE; n = 8 dogs/group. There were no significant differences among the groups.

    Article Snippet: Differences (P2 − P1 and P3 − P1) were calculated to assess the specific effect of GLP-1 over the effect achieved by pancreatic hormones and hyperglycemia per se.

    Techniques:

    GLP-1R-dependent pathway and GLP-1(9–36)-related pathway are involved in effects of GLP-1 on eNOS levels in HUVECs. (A) GLP-1R and DPP-4 proteins were detected in HUVECs by Western blot analysis. Cells were incubated in the presence or absence (control) of GLP-1, GLP-1(9–36) or exenatide (GLP-1R agonist) (all at 5000 pmol/L) for the following times (B, C, D, E). Cells were incubated with GLP-1 in the presence of exendin (9–39) (G+E) or sitaglipin (G+S) or both (G+E+S) for the following times (F, G, H). (B, F) After 30-min incubation, eNOS activity was determined by NO content in cells. (C, G) After 5-min incubation, phosphorylation of eNOS at ser-1177 was examined by Western blot analysis. (D, H) After 48-h incubation, total eNOS protein level was measured by Western blot analysis. The upper parts of C, D, G, and H show representative experiments. Data are mean±SD after normalization to β-actin level. (E) After 48-h incubation, eNOS mRNA level was quantified by real-time RT-PCR. Data are mean±SD after normalization to β-actin level. b P

    Journal: Acta Pharmacologica Sinica

    Article Title: Glucagon-like peptide-1 activates endothelial nitric oxide synthase in human umbilical vein endothelial cells

    doi: 10.1038/aps.2011.149

    Figure Lengend Snippet: GLP-1R-dependent pathway and GLP-1(9–36)-related pathway are involved in effects of GLP-1 on eNOS levels in HUVECs. (A) GLP-1R and DPP-4 proteins were detected in HUVECs by Western blot analysis. Cells were incubated in the presence or absence (control) of GLP-1, GLP-1(9–36) or exenatide (GLP-1R agonist) (all at 5000 pmol/L) for the following times (B, C, D, E). Cells were incubated with GLP-1 in the presence of exendin (9–39) (G+E) or sitaglipin (G+S) or both (G+E+S) for the following times (F, G, H). (B, F) After 30-min incubation, eNOS activity was determined by NO content in cells. (C, G) After 5-min incubation, phosphorylation of eNOS at ser-1177 was examined by Western blot analysis. (D, H) After 48-h incubation, total eNOS protein level was measured by Western blot analysis. The upper parts of C, D, G, and H show representative experiments. Data are mean±SD after normalization to β-actin level. (E) After 48-h incubation, eNOS mRNA level was quantified by real-time RT-PCR. Data are mean±SD after normalization to β-actin level. b P

    Article Snippet: Taken together, our results suggest that GLP-1 upregulates eNOS activity and protein expression through the GLP-1R-dependent and GLP-1(9–36)-related pathways in HUVECs.

    Techniques: Western Blot, Incubation, Activity Assay, Quantitative RT-PCR

    GLP-1 promotes endothelial nitric oxide synthase (eNOS) activity in human umbilical vein endothelial cells (HUVECs). HUVECs were incubated for 30 min in the absence or presence of GLP-1 (5–5000 pmol/L). Nitric oxide production was assayed using the fluorescent probe DAF-FM in the presence or absence of L -NAME (1 mmol/L). The experiment was repeated 3 times, and the values are means±SD. b P

    Journal: Acta Pharmacologica Sinica

    Article Title: Glucagon-like peptide-1 activates endothelial nitric oxide synthase in human umbilical vein endothelial cells

    doi: 10.1038/aps.2011.149

    Figure Lengend Snippet: GLP-1 promotes endothelial nitric oxide synthase (eNOS) activity in human umbilical vein endothelial cells (HUVECs). HUVECs were incubated for 30 min in the absence or presence of GLP-1 (5–5000 pmol/L). Nitric oxide production was assayed using the fluorescent probe DAF-FM in the presence or absence of L -NAME (1 mmol/L). The experiment was repeated 3 times, and the values are means±SD. b P

    Article Snippet: Taken together, our results suggest that GLP-1 upregulates eNOS activity and protein expression through the GLP-1R-dependent and GLP-1(9–36)-related pathways in HUVECs.

    Techniques: Activity Assay, Incubation

    Effects of GLP-1 on eNOS Ser-1177 phosphorylation, mRNA and protein expression in HUVECs. (A) Cells were incubated in the presence of GLP-1 (5000 pmol/L) for 0 (control), 5, 10, and 30 min to investigate the time course of the effect on eNOS phosphorylation. (B) Cells were incubated in the absence (control) or presence of GLP-1 (5–5000 pmol/L) for 5 min to investigate the dose-dependence of the effect. eNOS phosphorylated at ser-1177 (P-eNOS 1177) was examined by Western blot analysis. Representative experiments are both shown in the upper parts. Band intensities, after normalization to β-actin, are expressed as ratios of control. Mean±SD. n =4. b P

    Journal: Acta Pharmacologica Sinica

    Article Title: Glucagon-like peptide-1 activates endothelial nitric oxide synthase in human umbilical vein endothelial cells

    doi: 10.1038/aps.2011.149

    Figure Lengend Snippet: Effects of GLP-1 on eNOS Ser-1177 phosphorylation, mRNA and protein expression in HUVECs. (A) Cells were incubated in the presence of GLP-1 (5000 pmol/L) for 0 (control), 5, 10, and 30 min to investigate the time course of the effect on eNOS phosphorylation. (B) Cells were incubated in the absence (control) or presence of GLP-1 (5–5000 pmol/L) for 5 min to investigate the dose-dependence of the effect. eNOS phosphorylated at ser-1177 (P-eNOS 1177) was examined by Western blot analysis. Representative experiments are both shown in the upper parts. Band intensities, after normalization to β-actin, are expressed as ratios of control. Mean±SD. n =4. b P

    Article Snippet: Taken together, our results suggest that GLP-1 upregulates eNOS activity and protein expression through the GLP-1R-dependent and GLP-1(9–36)-related pathways in HUVECs.

    Techniques: Expressing, Incubation, Western Blot

    Expression and abundance of GLP-1R and GIPR in monocytes/macrophages and arterial SMCs. a Glp1r and ( b ) Gipr mRNA in exudate peritoneal macrophages and tissues as labelled from non-treated Apoe −/− mice were measured by real-time RT-PCR. c Western blotting analyses of GLP-1R and ( d ) GIPR abundance in exudate peritoneal macrophages of non-treated Apoe −/− mice and other mouse tissues as labelled. Three independent experiments were performed. e Immunostaining of GLP-1R or GIPR in exudate peritoneal macrophages from non-treated Apoe −/− mice. Green, GLP-1R or GIPR as indicated; red, nuclei; red + green, overlay of GLP-1R or GIPR and nuclei. Scale bars 50 μm. f GLP1R and ( g ) GIPR mRNA levels in human coronary artery SMCs, THP1 cells and THP1-derived macrophages. h GIPR mRNA level in human monocytes and monocyte-derived macrophages were measured by real-time RT-PCR. Results were obtained from three to six independent experiments

    Journal: Diabetologia

    Article Title: Native incretins prevent the development of atherosclerotic lesions in apolipoprotein E knockout mice

    doi: 10.1007/s00125-011-2241-2

    Figure Lengend Snippet: Expression and abundance of GLP-1R and GIPR in monocytes/macrophages and arterial SMCs. a Glp1r and ( b ) Gipr mRNA in exudate peritoneal macrophages and tissues as labelled from non-treated Apoe −/− mice were measured by real-time RT-PCR. c Western blotting analyses of GLP-1R and ( d ) GIPR abundance in exudate peritoneal macrophages of non-treated Apoe −/− mice and other mouse tissues as labelled. Three independent experiments were performed. e Immunostaining of GLP-1R or GIPR in exudate peritoneal macrophages from non-treated Apoe −/− mice. Green, GLP-1R or GIPR as indicated; red, nuclei; red + green, overlay of GLP-1R or GIPR and nuclei. Scale bars 50 μm. f GLP1R and ( g ) GIPR mRNA levels in human coronary artery SMCs, THP1 cells and THP1-derived macrophages. h GIPR mRNA level in human monocytes and monocyte-derived macrophages were measured by real-time RT-PCR. Results were obtained from three to six independent experiments

    Article Snippet: Our in vitro study showed that the suppressive effects of active forms of GLP-1 and GIP on macrophage foam cell formation were abolished by specific antagonists of GLP-1R and GIPR.

    Techniques: Expressing, Mouse Assay, Quantitative RT-PCR, Western Blot, Immunostaining, Derivative Assay

    GLP1 restored the attenuated b-catenin signaling induced by PA. (A) Representative images of subcellular distribution of active b-catenin (red, b-catenin and blue, DAPI; scale bar, 20 μm) in cardiomyocytes incubated with PA in the absence or presence of GLP1 (25 nM). (B and C) Western blot assay for cytosolic and nuclear b-catenin, survivin and BCL2. Intensities were quantified and normalized against the level of GAPDH or histone-3 and expressed as fold changes of protein abundance under PA stimulus. Data are means± s.e.m . of three independent experiments. * P

    Journal: Journal of Molecular Endocrinology

    Article Title: GLP1 protects cardiomyocytes from palmitate-induced apoptosis via Akt/GSK3b/b-catenin pathway

    doi: 10.1530/JME-15-0155

    Figure Lengend Snippet: GLP1 restored the attenuated b-catenin signaling induced by PA. (A) Representative images of subcellular distribution of active b-catenin (red, b-catenin and blue, DAPI; scale bar, 20 μm) in cardiomyocytes incubated with PA in the absence or presence of GLP1 (25 nM). (B and C) Western blot assay for cytosolic and nuclear b-catenin, survivin and BCL2. Intensities were quantified and normalized against the level of GAPDH or histone-3 and expressed as fold changes of protein abundance under PA stimulus. Data are means± s.e.m . of three independent experiments. * P

    Article Snippet: b-Catenin was required for the anti-apoptotic effect of GLP1 in response to palmitate stress To further determine whether b-catenin is required for the anti-apoptotic effect of GLP1 on cardiomyocytes, we adopted a shRNA approach to knockdown b-catenin.

    Techniques: Incubation, Western Blot

    b-Catenin was required for GLP1-mediated anti-apoptotic effects upon lipotoxicity. A recombinant adenovirus coding shRNA for b-catenin (Ad-sh-b-catenin) or for scramble sequences (Ad-sh-sc) was constructed to express ZsGreen protein as a marker for the identification of infected cells. Cultured cardiomyocytes were infected with respective adenoviruses at a MOI of 10. After 24 h, cells were treated with palmitate (PA) and GLP1 (25 nM) for another 24 h. (A) Transduction efficiency of recombinant adenoviruses was assayed by ZsGreen fluorescence analysis (green, ZsGreen and blue, DAPI; scale bar, 40 μm) 1 day after infection. (B) The suppression efficiency of b-catenin using shRNA was determined by western blot analysis for total b-catenin expression. (C, D, E and F) The effect of b-catenin silencing on GLP1 action in PA-treated cardiomyocytes was further assessed. (C) Representative images showed TUNEL staining for apoptotic cells (red, TUNEL and blue, DAPI; green, ZsGreen; scale bar, 40 μm). (D) Quantification of apoptotic nuclei was expressed as the percentage of TUNEL-positive to DAPI-positive cells. (E and F) Western blot analysis for cleaved caspase-3, survivin and BCL2. Intensities of protein expression were quantified, normalized against the level of GAPDH and expressed as relative changes to protein abundance in cardiomyocytes infected with scramble control (Ad-sh-sc). Data are means± s.e.m . of three independent experiments. ** P

    Journal: Journal of Molecular Endocrinology

    Article Title: GLP1 protects cardiomyocytes from palmitate-induced apoptosis via Akt/GSK3b/b-catenin pathway

    doi: 10.1530/JME-15-0155

    Figure Lengend Snippet: b-Catenin was required for GLP1-mediated anti-apoptotic effects upon lipotoxicity. A recombinant adenovirus coding shRNA for b-catenin (Ad-sh-b-catenin) or for scramble sequences (Ad-sh-sc) was constructed to express ZsGreen protein as a marker for the identification of infected cells. Cultured cardiomyocytes were infected with respective adenoviruses at a MOI of 10. After 24 h, cells were treated with palmitate (PA) and GLP1 (25 nM) for another 24 h. (A) Transduction efficiency of recombinant adenoviruses was assayed by ZsGreen fluorescence analysis (green, ZsGreen and blue, DAPI; scale bar, 40 μm) 1 day after infection. (B) The suppression efficiency of b-catenin using shRNA was determined by western blot analysis for total b-catenin expression. (C, D, E and F) The effect of b-catenin silencing on GLP1 action in PA-treated cardiomyocytes was further assessed. (C) Representative images showed TUNEL staining for apoptotic cells (red, TUNEL and blue, DAPI; green, ZsGreen; scale bar, 40 μm). (D) Quantification of apoptotic nuclei was expressed as the percentage of TUNEL-positive to DAPI-positive cells. (E and F) Western blot analysis for cleaved caspase-3, survivin and BCL2. Intensities of protein expression were quantified, normalized against the level of GAPDH and expressed as relative changes to protein abundance in cardiomyocytes infected with scramble control (Ad-sh-sc). Data are means± s.e.m . of three independent experiments. ** P

    Article Snippet: b-Catenin was required for the anti-apoptotic effect of GLP1 in response to palmitate stress To further determine whether b-catenin is required for the anti-apoptotic effect of GLP1 on cardiomyocytes, we adopted a shRNA approach to knockdown b-catenin.

    Techniques: Recombinant, shRNA, Construct, Marker, Infection, Cell Culture, Transduction, Fluorescence, Western Blot, Expressing, TUNEL Assay, Staining

    Schematic of a proposed model for GLP1-mediated signaling pathway protecting cardiomyocytes from PA-induced lipotoxicity. PA, palmitate.

    Journal: Journal of Molecular Endocrinology

    Article Title: GLP1 protects cardiomyocytes from palmitate-induced apoptosis via Akt/GSK3b/b-catenin pathway

    doi: 10.1530/JME-15-0155

    Figure Lengend Snippet: Schematic of a proposed model for GLP1-mediated signaling pathway protecting cardiomyocytes from PA-induced lipotoxicity. PA, palmitate.

    Article Snippet: b-Catenin was required for the anti-apoptotic effect of GLP1 in response to palmitate stress To further determine whether b-catenin is required for the anti-apoptotic effect of GLP1 on cardiomyocytes, we adopted a shRNA approach to knockdown b-catenin.

    Techniques:

    GLP1 prevented CD36-mediated intracellular lipid accumulation in PA-stressed cardiomyocytes via the GLP1R/Akt axis. Cardiomyocytes were incubated with PA (400 μM) in the absence or presence of GLP1 (25 nM) alone, or in combination of GLP1 with Exd(9–39) (100 nM) or MK2206 (50 nM). (A) Oil red O staining for intracellular neutral lipids (red–brown, indicated by white arrow) accumulated in cardiomyocytes after the respective 12 h (top) or 24 h (bottom) incubation. Nuclei were counterstained by DAPI (blue). Scale bar represents 10 μm. (B) Subcellular distribution of CD36 (green, CD36; scale bar, 10 μm) in cardiomyocytes after 6 h incubation was examined by indirect immunofluorescence. (C) Levels of plasma membrane, cytosolic and total CD36 were determined by western blot. Intensities were quantified and normalized against the level of total CD36. Data are means± s.e.m . of three independent experiments. * P

    Journal: Journal of Molecular Endocrinology

    Article Title: GLP1 protects cardiomyocytes from palmitate-induced apoptosis via Akt/GSK3b/b-catenin pathway

    doi: 10.1530/JME-15-0155

    Figure Lengend Snippet: GLP1 prevented CD36-mediated intracellular lipid accumulation in PA-stressed cardiomyocytes via the GLP1R/Akt axis. Cardiomyocytes were incubated with PA (400 μM) in the absence or presence of GLP1 (25 nM) alone, or in combination of GLP1 with Exd(9–39) (100 nM) or MK2206 (50 nM). (A) Oil red O staining for intracellular neutral lipids (red–brown, indicated by white arrow) accumulated in cardiomyocytes after the respective 12 h (top) or 24 h (bottom) incubation. Nuclei were counterstained by DAPI (blue). Scale bar represents 10 μm. (B) Subcellular distribution of CD36 (green, CD36; scale bar, 10 μm) in cardiomyocytes after 6 h incubation was examined by indirect immunofluorescence. (C) Levels of plasma membrane, cytosolic and total CD36 were determined by western blot. Intensities were quantified and normalized against the level of total CD36. Data are means± s.e.m . of three independent experiments. * P

    Article Snippet: b-Catenin was required for the anti-apoptotic effect of GLP1 in response to palmitate stress To further determine whether b-catenin is required for the anti-apoptotic effect of GLP1 on cardiomyocytes, we adopted a shRNA approach to knockdown b-catenin.

    Techniques: Incubation, Staining, Immunofluorescence, Western Blot

    GLP1-mediated anti-apoptotic effects were abolished when Akt was inhibited. Cultured cardiomyocytes were incubated with PA (400 μM) for 24 h in the absence or presence of GLP1 (25 nM) alone, or in combination of GLP1 (25 nM) with an Akt inhibitor MK2206 (50 nM). (A) Apoptotic cardiomyocytes were examined by TUNEL staining (red, TUNEL and blue, DAPI; scale bar, 50 μm). (B) Numbers of apoptotic cells were quantified and expressed as the percentage of TUNEL-positive to DAPI-positive cells. (C) Levels of cleaved caspase-3 were analyzed by western blot and quantified by densitometry. Distribution of active b-catenin in cardiomyocytes was determined by immunostainning (red, b-catenin and blue, DAPI; scale bar, 10 μm) (D) and levels of cytosolic and nuclear b-catenin were further examined by western blot (E). Intensities were quantified and normalized against the level of GAPDH or histone-3 and expressed as fold changes of protein abundance under PA stimulus. Data are means± s.e.m . of three independent experiments. * P

    Journal: Journal of Molecular Endocrinology

    Article Title: GLP1 protects cardiomyocytes from palmitate-induced apoptosis via Akt/GSK3b/b-catenin pathway

    doi: 10.1530/JME-15-0155

    Figure Lengend Snippet: GLP1-mediated anti-apoptotic effects were abolished when Akt was inhibited. Cultured cardiomyocytes were incubated with PA (400 μM) for 24 h in the absence or presence of GLP1 (25 nM) alone, or in combination of GLP1 (25 nM) with an Akt inhibitor MK2206 (50 nM). (A) Apoptotic cardiomyocytes were examined by TUNEL staining (red, TUNEL and blue, DAPI; scale bar, 50 μm). (B) Numbers of apoptotic cells were quantified and expressed as the percentage of TUNEL-positive to DAPI-positive cells. (C) Levels of cleaved caspase-3 were analyzed by western blot and quantified by densitometry. Distribution of active b-catenin in cardiomyocytes was determined by immunostainning (red, b-catenin and blue, DAPI; scale bar, 10 μm) (D) and levels of cytosolic and nuclear b-catenin were further examined by western blot (E). Intensities were quantified and normalized against the level of GAPDH or histone-3 and expressed as fold changes of protein abundance under PA stimulus. Data are means± s.e.m . of three independent experiments. * P

    Article Snippet: b-Catenin was required for the anti-apoptotic effect of GLP1 in response to palmitate stress To further determine whether b-catenin is required for the anti-apoptotic effect of GLP1 on cardiomyocytes, we adopted a shRNA approach to knockdown b-catenin.

    Techniques: Cell Culture, Incubation, TUNEL Assay, Staining, Western Blot

    Reversal of b-catenin signaling by GLP1 was mediated via GLP1R/Akt/GSK3b/ pathways in PA-stressed cardiomyocytes. Cardiomyocytes were incubated with PA (400 μM) for 24 h in the absence or presence of GLP1 (25 nM) alone, or in combination of GLP1 with Exd(9–39) (100 nM) or MK2206 (50 nM). (A and B) Western blot analysis for GLP1R, phosphorylated-Akt (Ser473), total Akt, phosphorylated-GSK3b (Ser9), total GSK3b, active b-catenin, phosphorylated-b-catenin (Ser33/37/Thr41), phosphorylated-b-catenin (Ser552), survivin, BCL2 and GAPDH. Intensities were quantified and normalized against the level of total proteins (Akt and GSK3b) or GAPDH. Data are means± s.e.m . of three independent experiments. * P

    Journal: Journal of Molecular Endocrinology

    Article Title: GLP1 protects cardiomyocytes from palmitate-induced apoptosis via Akt/GSK3b/b-catenin pathway

    doi: 10.1530/JME-15-0155

    Figure Lengend Snippet: Reversal of b-catenin signaling by GLP1 was mediated via GLP1R/Akt/GSK3b/ pathways in PA-stressed cardiomyocytes. Cardiomyocytes were incubated with PA (400 μM) for 24 h in the absence or presence of GLP1 (25 nM) alone, or in combination of GLP1 with Exd(9–39) (100 nM) or MK2206 (50 nM). (A and B) Western blot analysis for GLP1R, phosphorylated-Akt (Ser473), total Akt, phosphorylated-GSK3b (Ser9), total GSK3b, active b-catenin, phosphorylated-b-catenin (Ser33/37/Thr41), phosphorylated-b-catenin (Ser552), survivin, BCL2 and GAPDH. Intensities were quantified and normalized against the level of total proteins (Akt and GSK3b) or GAPDH. Data are means± s.e.m . of three independent experiments. * P

    Article Snippet: b-Catenin was required for the anti-apoptotic effect of GLP1 in response to palmitate stress To further determine whether b-catenin is required for the anti-apoptotic effect of GLP1 on cardiomyocytes, we adopted a shRNA approach to knockdown b-catenin.

    Techniques: Incubation, Western Blot

    GLP1 antagonized palmitate-induced apoptosis of cardiomyocytes. Isolated cardiomyocytes were incubated with PA for 24 h in the absence or presence of different concentrations of GLP1 (10–50 nM). (A) Representative immunostaining for TUNEL-positive (red) cells. Nuclei were labeled with DAPI (blue). Scale bar represents 40 μm. (B) Apoptotic cells were quantified and expressed as the percentage of TUNEL-positive cells to DAPI-positive cells. (C) Western blot analysis of cleaved caspase-3 and cleaved-PARP in cardiomyocytes without or with GLP1 (25 nM) treatment under PA stress. Intensities were quantified and normalized against the level of GAPDH and expressed as percentage of protein abundance under PA stimulus. Data are means± s.e.m . of three independent experiments. * P

    Journal: Journal of Molecular Endocrinology

    Article Title: GLP1 protects cardiomyocytes from palmitate-induced apoptosis via Akt/GSK3b/b-catenin pathway

    doi: 10.1530/JME-15-0155

    Figure Lengend Snippet: GLP1 antagonized palmitate-induced apoptosis of cardiomyocytes. Isolated cardiomyocytes were incubated with PA for 24 h in the absence or presence of different concentrations of GLP1 (10–50 nM). (A) Representative immunostaining for TUNEL-positive (red) cells. Nuclei were labeled with DAPI (blue). Scale bar represents 40 μm. (B) Apoptotic cells were quantified and expressed as the percentage of TUNEL-positive cells to DAPI-positive cells. (C) Western blot analysis of cleaved caspase-3 and cleaved-PARP in cardiomyocytes without or with GLP1 (25 nM) treatment under PA stress. Intensities were quantified and normalized against the level of GAPDH and expressed as percentage of protein abundance under PA stimulus. Data are means± s.e.m . of three independent experiments. * P

    Article Snippet: b-Catenin was required for the anti-apoptotic effect of GLP1 in response to palmitate stress To further determine whether b-catenin is required for the anti-apoptotic effect of GLP1 on cardiomyocytes, we adopted a shRNA approach to knockdown b-catenin.

    Techniques: Isolation, Incubation, Immunostaining, TUNEL Assay, Labeling, Western Blot

    The anti-apoptotic effect of GLP1 occurred via the GLP1 receptor. (A and B) Isolated cardiomyocytes cultured at PA (400 μM) for 24 h were treated with GLP1 (25 nM) alone, or in combination with an increasing amount (25–400 nM) of Exd(9–39). Representative diagrams showed TUNEL staining of apoptotic cells (red, TUNEL and blue, DAPI; scale bar, 50 μm) (A). Quantification of apoptotic nuclei were expressed as the percentage of TUNEL-positive cells to DAPI-positive cells (B). 25 nM of GLP1 and 100 nM of Exd(9–39) were adopted in subsequent experiments (C, D, E and F). Western blot analysis for cleaved caspase-3 and cleaved-PARP (C); cytosolic and nuclear b-catenin (E); survivin and BCL2 (F). Intensities were quantified and normalized against the level of GAPDH or histone-3 and expressed as fold changes of protein abundance under PA stimulus. Data are means± s.e.m . of three independent experiments. * P

    Journal: Journal of Molecular Endocrinology

    Article Title: GLP1 protects cardiomyocytes from palmitate-induced apoptosis via Akt/GSK3b/b-catenin pathway

    doi: 10.1530/JME-15-0155

    Figure Lengend Snippet: The anti-apoptotic effect of GLP1 occurred via the GLP1 receptor. (A and B) Isolated cardiomyocytes cultured at PA (400 μM) for 24 h were treated with GLP1 (25 nM) alone, or in combination with an increasing amount (25–400 nM) of Exd(9–39). Representative diagrams showed TUNEL staining of apoptotic cells (red, TUNEL and blue, DAPI; scale bar, 50 μm) (A). Quantification of apoptotic nuclei were expressed as the percentage of TUNEL-positive cells to DAPI-positive cells (B). 25 nM of GLP1 and 100 nM of Exd(9–39) were adopted in subsequent experiments (C, D, E and F). Western blot analysis for cleaved caspase-3 and cleaved-PARP (C); cytosolic and nuclear b-catenin (E); survivin and BCL2 (F). Intensities were quantified and normalized against the level of GAPDH or histone-3 and expressed as fold changes of protein abundance under PA stimulus. Data are means± s.e.m . of three independent experiments. * P

    Article Snippet: b-Catenin was required for the anti-apoptotic effect of GLP1 in response to palmitate stress To further determine whether b-catenin is required for the anti-apoptotic effect of GLP1 on cardiomyocytes, we adopted a shRNA approach to knockdown b-catenin.

    Techniques: Isolation, Cell Culture, TUNEL Assay, Staining, Western Blot

    GLP-1 formulated in PGC-C18 has extended blood circulation time compared to GLP-1 alone

    Journal: Pharmaceutical research

    Article Title: Extending residence time and stability of peptides by Protected Graft Copolymer (PGC) excipient: GLP-1 example

    doi: 10.1007/s11095-011-0542-2

    Figure Lengend Snippet: GLP-1 formulated in PGC-C18 has extended blood circulation time compared to GLP-1 alone

    Article Snippet: An additional HPLC Gel Permeation Column for determination of PGC binding to GLP-1 (0.78 × 30 cm; BioSEP S2000 column) plus a reverse phase column (Synergi 2.5um Max, 0.4×2cm) for quantification of GLP-1 were from Phenomenex (Torrance, CA).

    Techniques: Pyrolysis Gas Chromatography

    Gel Permeation HPLC chromatograms showing the extent of binding of GLP-1 to various concentrations of PGC-C18 carrier

    Journal: Pharmaceutical research

    Article Title: Extending residence time and stability of peptides by Protected Graft Copolymer (PGC) excipient: GLP-1 example

    doi: 10.1007/s11095-011-0542-2

    Figure Lengend Snippet: Gel Permeation HPLC chromatograms showing the extent of binding of GLP-1 to various concentrations of PGC-C18 carrier

    Article Snippet: An additional HPLC Gel Permeation Column for determination of PGC binding to GLP-1 (0.78 × 30 cm; BioSEP S2000 column) plus a reverse phase column (Synergi 2.5um Max, 0.4×2cm) for quantification of GLP-1 were from Phenomenex (Torrance, CA).

    Techniques: High Performance Liquid Chromatography, Binding Assay, Pyrolysis Gas Chromatography

    Scatchard plot of GLP-1 binding to PGC-C18 shows various affinities

    Journal: Pharmaceutical research

    Article Title: Extending residence time and stability of peptides by Protected Graft Copolymer (PGC) excipient: GLP-1 example

    doi: 10.1007/s11095-011-0542-2

    Figure Lengend Snippet: Scatchard plot of GLP-1 binding to PGC-C18 shows various affinities

    Article Snippet: An additional HPLC Gel Permeation Column for determination of PGC binding to GLP-1 (0.78 × 30 cm; BioSEP S2000 column) plus a reverse phase column (Synergi 2.5um Max, 0.4×2cm) for quantification of GLP-1 were from Phenomenex (Torrance, CA).

    Techniques: Binding Assay, Pyrolysis Gas Chromatography

    PGC with C18 protects GLP-1 from rapid DPP IV digestion but not PGC without C18

    Journal: Pharmaceutical research

    Article Title: Extending residence time and stability of peptides by Protected Graft Copolymer (PGC) excipient: GLP-1 example

    doi: 10.1007/s11095-011-0542-2

    Figure Lengend Snippet: PGC with C18 protects GLP-1 from rapid DPP IV digestion but not PGC without C18

    Article Snippet: An additional HPLC Gel Permeation Column for determination of PGC binding to GLP-1 (0.78 × 30 cm; BioSEP S2000 column) plus a reverse phase column (Synergi 2.5um Max, 0.4×2cm) for quantification of GLP-1 were from Phenomenex (Torrance, CA).

    Techniques: Pyrolysis Gas Chromatography

    GLP-1 formulated in PGC-C18 is effective in improving HbA1c level in ZDF rats

    Journal: Pharmaceutical research

    Article Title: Extending residence time and stability of peptides by Protected Graft Copolymer (PGC) excipient: GLP-1 example

    doi: 10.1007/s11095-011-0542-2

    Figure Lengend Snippet: GLP-1 formulated in PGC-C18 is effective in improving HbA1c level in ZDF rats

    Article Snippet: An additional HPLC Gel Permeation Column for determination of PGC binding to GLP-1 (0.78 × 30 cm; BioSEP S2000 column) plus a reverse phase column (Synergi 2.5um Max, 0.4×2cm) for quantification of GLP-1 were from Phenomenex (Torrance, CA).

    Techniques: Pyrolysis Gas Chromatography

    Calcium influx experiment in INS-1 cells indicates that GLP-1 formulated in PGC-C18 is biologically active

    Journal: Pharmaceutical research

    Article Title: Extending residence time and stability of peptides by Protected Graft Copolymer (PGC) excipient: GLP-1 example

    doi: 10.1007/s11095-011-0542-2

    Figure Lengend Snippet: Calcium influx experiment in INS-1 cells indicates that GLP-1 formulated in PGC-C18 is biologically active

    Article Snippet: An additional HPLC Gel Permeation Column for determination of PGC binding to GLP-1 (0.78 × 30 cm; BioSEP S2000 column) plus a reverse phase column (Synergi 2.5um Max, 0.4×2cm) for quantification of GLP-1 were from Phenomenex (Torrance, CA).

    Techniques: Pyrolysis Gas Chromatography

    ffar2 and ffar3 knockout mice have impaired glucose tolerance. A : Glucose stimulated GLP-1 secretion in vivo. ffar2 −/− and ffar3 −/− mice and wild-type littermates ( n = 5 each) were dosed with DPP4 inhibitor at a dose of 20 mg/kg per os after a 4-h fast. Thirty minutes post–DPP4 inhibitor dosing, mice were dosed with 1.5 g/kg glucose per os Plasma active GLP-1 was assessed by a MesoScale assay at 0 and 30 min of the oral glucose tolerance test. Data represent means ± 1 SEM, and statistical significance was assessed by Student t test: * P

    Journal: Diabetes

    Article Title: Short-Chain Fatty Acids Stimulate Glucagon-Like Peptide-1 Secretion via the G-Protein-Coupled Receptor FFAR2

    doi: 10.2337/db11-1019

    Figure Lengend Snippet: ffar2 and ffar3 knockout mice have impaired glucose tolerance. A : Glucose stimulated GLP-1 secretion in vivo. ffar2 −/− and ffar3 −/− mice and wild-type littermates ( n = 5 each) were dosed with DPP4 inhibitor at a dose of 20 mg/kg per os after a 4-h fast. Thirty minutes post–DPP4 inhibitor dosing, mice were dosed with 1.5 g/kg glucose per os Plasma active GLP-1 was assessed by a MesoScale assay at 0 and 30 min of the oral glucose tolerance test. Data represent means ± 1 SEM, and statistical significance was assessed by Student t test: * P

    Article Snippet: No explanation was found for the increased food intake, but our data suggest that it may be related to reduced secretion of L- cell peptides like GLP-1 and PYY.

    Techniques: Knock-Out, Mouse Assay, In Vivo

    ffar2 and ffar3 knockout impairs SCFA-triggered GLP-1 secretion. A : GLP-1 secretion from primary colonic cultures from wild-type, ffar2 −/− , and ffar3 −/− mice. Mixed primary cultures from the colon from wild-type, ffar3 −/− , and ffar2 −/− mice were incubated in bath solution containing 10 mmol/L glucose together with acetate (1 mmol/L), propionate (1 mmol/L), and IBMX (100 μmol/L) as indicated (all n = 6). B : GLP-1 secretion from primary colonic cultures from wild-type, ffar2 −/− , and ffar3 −/− mice triggered by a 140 mmol/L cocktail of SCFAs and an osmotic control of 140 mmol/L NaCl. GLP-1 secretion in each well is expressed relative to the basal secretion (control) measured in parallel on the same day, and error bars represent 1 SEM. Effects of SCFAs in the absence (* P

    Journal: Diabetes

    Article Title: Short-Chain Fatty Acids Stimulate Glucagon-Like Peptide-1 Secretion via the G-Protein-Coupled Receptor FFAR2

    doi: 10.2337/db11-1019

    Figure Lengend Snippet: ffar2 and ffar3 knockout impairs SCFA-triggered GLP-1 secretion. A : GLP-1 secretion from primary colonic cultures from wild-type, ffar2 −/− , and ffar3 −/− mice. Mixed primary cultures from the colon from wild-type, ffar3 −/− , and ffar2 −/− mice were incubated in bath solution containing 10 mmol/L glucose together with acetate (1 mmol/L), propionate (1 mmol/L), and IBMX (100 μmol/L) as indicated (all n = 6). B : GLP-1 secretion from primary colonic cultures from wild-type, ffar2 −/− , and ffar3 −/− mice triggered by a 140 mmol/L cocktail of SCFAs and an osmotic control of 140 mmol/L NaCl. GLP-1 secretion in each well is expressed relative to the basal secretion (control) measured in parallel on the same day, and error bars represent 1 SEM. Effects of SCFAs in the absence (* P

    Article Snippet: No explanation was found for the increased food intake, but our data suggest that it may be related to reduced secretion of L- cell peptides like GLP-1 and PYY.

    Techniques: Knock-Out, Mouse Assay, Incubation

    SCFAs stimulate GLP-1 secretion. A : Acute stimulation of GLP-1 secretion. Mixed primary cultures from murine colon were incubated for 2 h in 10 mmol/L glucose (Con) or in the additional presence of acetate (Ace) (1 mmol/L), propionate (Pro) (1 mmol/L), or butyrate (But) (1 mmol/L) with or without IBMX (100 μmol/L) as indicated. GLP-1 secretion in each well is expressed relative to the basal secretion (Con) measured in parallel on the same day. Data represent the means ± SEM of the number of wells indicated above each bar. * P

    Journal: Diabetes

    Article Title: Short-Chain Fatty Acids Stimulate Glucagon-Like Peptide-1 Secretion via the G-Protein-Coupled Receptor FFAR2

    doi: 10.2337/db11-1019

    Figure Lengend Snippet: SCFAs stimulate GLP-1 secretion. A : Acute stimulation of GLP-1 secretion. Mixed primary cultures from murine colon were incubated for 2 h in 10 mmol/L glucose (Con) or in the additional presence of acetate (Ace) (1 mmol/L), propionate (Pro) (1 mmol/L), or butyrate (But) (1 mmol/L) with or without IBMX (100 μmol/L) as indicated. GLP-1 secretion in each well is expressed relative to the basal secretion (Con) measured in parallel on the same day. Data represent the means ± SEM of the number of wells indicated above each bar. * P

    Article Snippet: No explanation was found for the increased food intake, but our data suggest that it may be related to reduced secretion of L- cell peptides like GLP-1 and PYY.

    Techniques: Incubation

    Propionate (Prop) responses are not sensitive to pertussis toxin (Ptx). A : GLP-1 secretion from primary colonic cultures treated with IBMX (100 μmol/L) with or without somatostatin (Sst) (100 nmol/L) in the absence (■) or presence (□) of 0.2 μg/mL pertussis toxin (all n = 3). B : GLP-1 secretion from primary colonic cultures triggered by propionate (1 mmol/L) in the absence and presence of pertussis toxin (0.2 μg/mL). The number of wells is indicated above the bars. Mixed primary cultures from the colon were incubated in bath solution containing reagents as indicated. GLP-1 secretion in each well is expressed relative to the basal secretion (control), measured in parallel on the same day. Data represent the means ± SEM of the number of wells indicated. Statistical significance was assessed by one-way ANOVA with a post hoc Bonferroni correction test: * P

    Journal: Diabetes

    Article Title: Short-Chain Fatty Acids Stimulate Glucagon-Like Peptide-1 Secretion via the G-Protein-Coupled Receptor FFAR2

    doi: 10.2337/db11-1019

    Figure Lengend Snippet: Propionate (Prop) responses are not sensitive to pertussis toxin (Ptx). A : GLP-1 secretion from primary colonic cultures treated with IBMX (100 μmol/L) with or without somatostatin (Sst) (100 nmol/L) in the absence (■) or presence (□) of 0.2 μg/mL pertussis toxin (all n = 3). B : GLP-1 secretion from primary colonic cultures triggered by propionate (1 mmol/L) in the absence and presence of pertussis toxin (0.2 μg/mL). The number of wells is indicated above the bars. Mixed primary cultures from the colon were incubated in bath solution containing reagents as indicated. GLP-1 secretion in each well is expressed relative to the basal secretion (control), measured in parallel on the same day. Data represent the means ± SEM of the number of wells indicated. Statistical significance was assessed by one-way ANOVA with a post hoc Bonferroni correction test: * P

    Article Snippet: No explanation was found for the increased food intake, but our data suggest that it may be related to reduced secretion of L- cell peptides like GLP-1 and PYY.

    Techniques: Incubation

    The mean activity of human gastric lipase (HGL) and mean concentrations of cholecystokinin (CCK), glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic peptide (GIP) in the study groups (non-HPG—non- Helicobacter pylori gastritis group, HPG— Helicobacter pylori gastritis group, CG—control group).

    Journal: Nutrients

    Article Title: Gastric Lipase Secretion in Children with Gastritis

    doi: 10.3390/nu5082924

    Figure Lengend Snippet: The mean activity of human gastric lipase (HGL) and mean concentrations of cholecystokinin (CCK), glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic peptide (GIP) in the study groups (non-HPG—non- Helicobacter pylori gastritis group, HPG— Helicobacter pylori gastritis group, CG—control group).

    Article Snippet: Wojdemann et al . and Borovicka et al . observed that GLP-1 and CCK inhibited the secretion of HGL [ , , ].

    Techniques: Activity Assay

    GLP-1 response during OGTT per model. Data are shown as mean ± SEM. P values are for the comparison of the AUC for each group.

    Journal: Journal of the Endocrine Society

    Article Title: Oral Glucose Tolerance Test Glucose Peak Time Is Most Predictive of Prediabetes and Hepatic Steatosis in Obese Girls

    doi: 10.1210/js.2018-00041

    Figure Lengend Snippet: GLP-1 response during OGTT per model. Data are shown as mean ± SEM. P values are for the comparison of the AUC for each group.

    Article Snippet: We measured GLP-1 and sampled at extra time points in the first part of the OGTT to better understand the early response to the glucose.

    Techniques:

    GLP-1 actions and target organs . GLP-1, through its receptor GLP-1R, has functional effects on a variety of tissues.

    Journal: Frontiers in Endocrinology

    Article Title: Beyond Glycemic Control in Diabetes Mellitus: Effects of Incretin-Based Therapies on Bone Metabolism

    doi: 10.3389/fendo.2013.00073

    Figure Lengend Snippet: GLP-1 actions and target organs . GLP-1, through its receptor GLP-1R, has functional effects on a variety of tissues.

    Article Snippet: This event may be mediated by gastrointestinal hormones released after meal ingestion such as GLP-1 and GIP (Henriksen et al., ).

    Techniques: Functional Assay

    Effects of incretin-based therapies on bone metabolism . GLP-1 receptor agonists and DPP-4 inhibitors (via endogenous GLP-1) may stimulate osteoblastogenesis and inhibit osteoclastogenesis. Specifically, osteoblastogenesis stimulation has been hypothesized to occur via activation of Wnt/beta-catenin pathway and/or increased OPG/RANKL ratio. In addition, osteoclastogenesis inhibition has been suggested to be mediated by reduced sclerostin levels. GLP-1 RA, GLP-1 receptor agonists; DPP4i, dipeptidyl peptidase-4 inhibitors; OPG, osteoprotegerin; RANKL, receptor activator of nuclear factor kappa-B ligand.

    Journal: Frontiers in Endocrinology

    Article Title: Beyond Glycemic Control in Diabetes Mellitus: Effects of Incretin-Based Therapies on Bone Metabolism

    doi: 10.3389/fendo.2013.00073

    Figure Lengend Snippet: Effects of incretin-based therapies on bone metabolism . GLP-1 receptor agonists and DPP-4 inhibitors (via endogenous GLP-1) may stimulate osteoblastogenesis and inhibit osteoclastogenesis. Specifically, osteoblastogenesis stimulation has been hypothesized to occur via activation of Wnt/beta-catenin pathway and/or increased OPG/RANKL ratio. In addition, osteoclastogenesis inhibition has been suggested to be mediated by reduced sclerostin levels. GLP-1 RA, GLP-1 receptor agonists; DPP4i, dipeptidyl peptidase-4 inhibitors; OPG, osteoprotegerin; RANKL, receptor activator of nuclear factor kappa-B ligand.

    Article Snippet: This event may be mediated by gastrointestinal hormones released after meal ingestion such as GLP-1 and GIP (Henriksen et al., ).

    Techniques: Activation Assay, Inhibition

    A and B, Effects of vildagliptin and GLP-1 given separately, GLP-1 in combination with vildagliptin, Ex-4, or saline on food intake in WT mice (A) and rats (B). Data are mean ± sem . *, P

    Journal: Endocrinology

    Article Title: Suppression of Food Intake by Glucagon-Like Peptide-1 Receptor Agonists: Relative Potencies and Role of Dipeptidyl Peptidase-4

    doi: 10.1210/en.2012-1358

    Figure Lengend Snippet: A and B, Effects of vildagliptin and GLP-1 given separately, GLP-1 in combination with vildagliptin, Ex-4, or saline on food intake in WT mice (A) and rats (B). Data are mean ± sem . *, P

    Article Snippet: To investigate whether targeted deletion of the mouse Dpp4 gene would increase the anorectic effect of GLP-1 compared with that of Ex-4, Dpp4 −/− mice received ip injections of GLP-1 (5, 50, and 500 μg), Ex-4 (1.25, 2.5, and 10 μg), or saline.

    Techniques: Mouse Assay

    Plasma clearance (milliliters per minute) of total GLP-1, intact GLP-1, and Ex-4 in rats pretreated with saline or vildagliptin. Because clearance of total GLP-1 and Ex-4 was similar after both the saline and vildagliptin treatments, the data were pooled.

    Journal: Endocrinology

    Article Title: Suppression of Food Intake by Glucagon-Like Peptide-1 Receptor Agonists: Relative Potencies and Role of Dipeptidyl Peptidase-4

    doi: 10.1210/en.2012-1358

    Figure Lengend Snippet: Plasma clearance (milliliters per minute) of total GLP-1, intact GLP-1, and Ex-4 in rats pretreated with saline or vildagliptin. Because clearance of total GLP-1 and Ex-4 was similar after both the saline and vildagliptin treatments, the data were pooled.

    Article Snippet: To investigate whether targeted deletion of the mouse Dpp4 gene would increase the anorectic effect of GLP-1 compared with that of Ex-4, Dpp4 −/− mice received ip injections of GLP-1 (5, 50, and 500 μg), Ex-4 (1.25, 2.5, and 10 μg), or saline.

    Techniques:

    A–C, Plasma concentrations of total GLP-1 (A), intact GLP-1(7–36NH 2 ) (B), and ratios of intact to total GLP-1 (C) in Dpp4 −/− and WT mice 30 min after injection 100 μg GLP-1; D, plasma DPP-4 activity measured in

    Journal: Endocrinology

    Article Title: Suppression of Food Intake by Glucagon-Like Peptide-1 Receptor Agonists: Relative Potencies and Role of Dipeptidyl Peptidase-4

    doi: 10.1210/en.2012-1358

    Figure Lengend Snippet: A–C, Plasma concentrations of total GLP-1 (A), intact GLP-1(7–36NH 2 ) (B), and ratios of intact to total GLP-1 (C) in Dpp4 −/− and WT mice 30 min after injection 100 μg GLP-1; D, plasma DPP-4 activity measured in

    Article Snippet: To investigate whether targeted deletion of the mouse Dpp4 gene would increase the anorectic effect of GLP-1 compared with that of Ex-4, Dpp4 −/− mice received ip injections of GLP-1 (5, 50, and 500 μg), Ex-4 (1.25, 2.5, and 10 μg), or saline.

    Techniques: Mouse Assay, Injection, Activity Assay

    A–C, Food intake after ip GLP-1 (500 μg) or saline alone or in combination with vildagliptin (150 μg) in WT mice (A); GLP-1 (5, 50, 500 μg) in WT (B) or Dpp4 −/− (C) mice. Data are mean ± sem . *,

    Journal: Endocrinology

    Article Title: Suppression of Food Intake by Glucagon-Like Peptide-1 Receptor Agonists: Relative Potencies and Role of Dipeptidyl Peptidase-4

    doi: 10.1210/en.2012-1358

    Figure Lengend Snippet: A–C, Food intake after ip GLP-1 (500 μg) or saline alone or in combination with vildagliptin (150 μg) in WT mice (A); GLP-1 (5, 50, 500 μg) in WT (B) or Dpp4 −/− (C) mice. Data are mean ± sem . *,

    Article Snippet: To investigate whether targeted deletion of the mouse Dpp4 gene would increase the anorectic effect of GLP-1 compared with that of Ex-4, Dpp4 −/− mice received ip injections of GLP-1 (5, 50, and 500 μg), Ex-4 (1.25, 2.5, and 10 μg), or saline.

    Techniques: Mouse Assay

    A and B, Food intake in rats after ip GLP-1 (10, 100, 500 μg) (A) or GLP-1 (10, 100, 500 μg) (B) in combination with vildagliptin (10 mg). Data are mean ± sem . *, P

    Journal: Endocrinology

    Article Title: Suppression of Food Intake by Glucagon-Like Peptide-1 Receptor Agonists: Relative Potencies and Role of Dipeptidyl Peptidase-4

    doi: 10.1210/en.2012-1358

    Figure Lengend Snippet: A and B, Food intake in rats after ip GLP-1 (10, 100, 500 μg) (A) or GLP-1 (10, 100, 500 μg) (B) in combination with vildagliptin (10 mg). Data are mean ± sem . *, P

    Article Snippet: To investigate whether targeted deletion of the mouse Dpp4 gene would increase the anorectic effect of GLP-1 compared with that of Ex-4, Dpp4 −/− mice received ip injections of GLP-1 (5, 50, and 500 μg), Ex-4 (1.25, 2.5, and 10 μg), or saline.

    Techniques:

    A, Plasma total GLP-1 response to Ensure in RYGB (n = 7) and sham rats (n = 7); B, food intake in RYGB and sham rats after ip injection of vildagliptin (10 mg) or saline. Data are mean ± sem .

    Journal: Endocrinology

    Article Title: Suppression of Food Intake by Glucagon-Like Peptide-1 Receptor Agonists: Relative Potencies and Role of Dipeptidyl Peptidase-4

    doi: 10.1210/en.2012-1358

    Figure Lengend Snippet: A, Plasma total GLP-1 response to Ensure in RYGB (n = 7) and sham rats (n = 7); B, food intake in RYGB and sham rats after ip injection of vildagliptin (10 mg) or saline. Data are mean ± sem .

    Article Snippet: To investigate whether targeted deletion of the mouse Dpp4 gene would increase the anorectic effect of GLP-1 compared with that of Ex-4, Dpp4 −/− mice received ip injections of GLP-1 (5, 50, and 500 μg), Ex-4 (1.25, 2.5, and 10 μg), or saline.

    Techniques: Injection

    Summary of single-fiber responses. A–E ) Response profiles of S-type (GLP-1-sensitive: n = 7, GLP-1-insensitive: n = 10), S-type after Exendin 4(3-39) treatment ( n = 9), electrolytes-responsive type (E-type, n = 13), NaCl-specific type (N-type,

    Journal: The FASEB Journal

    Article Title: Glucagon-like peptide-1 is specifically involved in sweet taste transmission

    doi: 10.1096/fj.14-265355

    Figure Lengend Snippet: Summary of single-fiber responses. A–E ) Response profiles of S-type (GLP-1-sensitive: n = 7, GLP-1-insensitive: n = 10), S-type after Exendin 4(3-39) treatment ( n = 9), electrolytes-responsive type (E-type, n = 13), NaCl-specific type (N-type,

    Article Snippet: To determine the number of cells expressing GAD67-GFP, GLP-1, and T1R3, we counted positive cells in each taste bud in horizontal sections of FP and CV (the number of taste buds; FP: 121 CV: 211).

    Techniques:

    The effect of intravenous injection of GLP-1. A ) GLP-1-sensitive and -insensitive S-type fibers. S-type fibers that elicited more than 30 impulses/10 s in response to i.v. injection of GLP-1 were classified as GLP-1-sensitive fibers. B ) Time-dependent

    Journal: The FASEB Journal

    Article Title: Glucagon-like peptide-1 is specifically involved in sweet taste transmission

    doi: 10.1096/fj.14-265355

    Figure Lengend Snippet: The effect of intravenous injection of GLP-1. A ) GLP-1-sensitive and -insensitive S-type fibers. S-type fibers that elicited more than 30 impulses/10 s in response to i.v. injection of GLP-1 were classified as GLP-1-sensitive fibers. B ) Time-dependent

    Article Snippet: To determine the number of cells expressing GAD67-GFP, GLP-1, and T1R3, we counted positive cells in each taste bud in horizontal sections of FP and CV (the number of taste buds; FP: 121 CV: 211).

    Techniques: Injection

    GLP-1 secretion from sweet-sensitive TCs. A ) GLP-1 secretion from single taste buds. Apical side of an individual taste bud was stimulated for 60 seconds with DW, saccharin (Sac; 5, 20, or 50 mM Sac), sucrose (Suc; 500 or 1000 mM Suc), glucose (GLC; 500

    Journal: The FASEB Journal

    Article Title: Glucagon-like peptide-1 is specifically involved in sweet taste transmission

    doi: 10.1096/fj.14-265355

    Figure Lengend Snippet: GLP-1 secretion from sweet-sensitive TCs. A ) GLP-1 secretion from single taste buds. Apical side of an individual taste bud was stimulated for 60 seconds with DW, saccharin (Sac; 5, 20, or 50 mM Sac), sucrose (Suc; 500 or 1000 mM Suc), glucose (GLC; 500

    Article Snippet: To determine the number of cells expressing GAD67-GFP, GLP-1, and T1R3, we counted positive cells in each taste bud in horizontal sections of FP and CV (the number of taste buds; FP: 121 CV: 211).

    Techniques: Gas Chromatography

    Presence of GLP-1 in TCs and GLP-1R in gustatory neurons. A ) Immunofluorescence of GAD67-GFP (green), GLP-1 (red), T1R3 (cyan), and merged images in mouse FP (upper) and CV (lower) papillae taste buds. B ) Quantitative diagrams of the overlapping distribution

    Journal: The FASEB Journal

    Article Title: Glucagon-like peptide-1 is specifically involved in sweet taste transmission

    doi: 10.1096/fj.14-265355

    Figure Lengend Snippet: Presence of GLP-1 in TCs and GLP-1R in gustatory neurons. A ) Immunofluorescence of GAD67-GFP (green), GLP-1 (red), T1R3 (cyan), and merged images in mouse FP (upper) and CV (lower) papillae taste buds. B ) Quantitative diagrams of the overlapping distribution

    Article Snippet: To determine the number of cells expressing GAD67-GFP, GLP-1, and T1R3, we counted positive cells in each taste bud in horizontal sections of FP and CV (the number of taste buds; FP: 121 CV: 211).

    Techniques: Immunofluorescence

    Sweet tastants evoke GLP-1 secretion from sweet-sensitive TCs

    Journal: The FASEB Journal

    Article Title: Glucagon-like peptide-1 is specifically involved in sweet taste transmission

    doi: 10.1096/fj.14-265355

    Figure Lengend Snippet: Sweet tastants evoke GLP-1 secretion from sweet-sensitive TCs

    Article Snippet: To determine the number of cells expressing GAD67-GFP, GLP-1, and T1R3, we counted positive cells in each taste bud in horizontal sections of FP and CV (the number of taste buds; FP: 121 CV: 211).

    Techniques:

    GLP-1 activates sweet-best S-type gustatory nerve fibers. A ) A representative recording from a sweet-best CT fiber responding to tastants and intravenous injection of GLP-1. Taste stimuli were 10 mM HCl, 20 mM QHCl, 100 mM NaCl, 100 mM MPG, and 500 mM

    Journal: The FASEB Journal

    Article Title: Glucagon-like peptide-1 is specifically involved in sweet taste transmission

    doi: 10.1096/fj.14-265355

    Figure Lengend Snippet: GLP-1 activates sweet-best S-type gustatory nerve fibers. A ) A representative recording from a sweet-best CT fiber responding to tastants and intravenous injection of GLP-1. Taste stimuli were 10 mM HCl, 20 mM QHCl, 100 mM NaCl, 100 mM MPG, and 500 mM

    Article Snippet: To determine the number of cells expressing GAD67-GFP, GLP-1, and T1R3, we counted positive cells in each taste bud in horizontal sections of FP and CV (the number of taste buds; FP: 121 CV: 211).

    Techniques: Injection

    Plasma proinsulin levels before and after bolus injections of 5 g arginine under both basal and approximately 230 mg/dl hyperglycemic clamp conditions. A, Results for the islet transplant group (n = 5) during GLP-1 7–36 amide (▪) or placebo (□) infusion. B, Results for the pancreas transplant group (n = 6) during GLP-1 7–36 amide (•) or placebo (○) infusion.

    Journal: The Journal of Clinical Endocrinology and Metabolism

    Article Title: Effect of Glucagon-Like Peptide-1 on ?- and ?-Cell Function in Isolated Islet and Whole Pancreas Transplant Recipients

    doi: 10.1210/jc.2008-1806

    Figure Lengend Snippet: Plasma proinsulin levels before and after bolus injections of 5 g arginine under both basal and approximately 230 mg/dl hyperglycemic clamp conditions. A, Results for the islet transplant group (n = 5) during GLP-1 7–36 amide (▪) or placebo (□) infusion. B, Results for the pancreas transplant group (n = 6) during GLP-1 7–36 amide (•) or placebo (○) infusion.

    Article Snippet: In the islet group, insulin levels were significantly greater after 30 min of GLP-1 compared to placebo infusion (21.6 ± 7.8 vs .

    Techniques:

    Plasma glucose levels before and 30 min after GLP-1 7–36 amide ( filled columns ) or placebo ( lined columns ) infusion, and at the end of the approximately 230 mg/dl hyperglycemic clamp. Against the opposite y-axis, M represents the glucose infusion rate required during the hyperglycemic clamp. A, Results for the islet transplant group (n = 5). B, Results for the pancreas transplant group (n = 6). To convert glucose to mmol/liter, multiply by 0.05551.

    Journal: The Journal of Clinical Endocrinology and Metabolism

    Article Title: Effect of Glucagon-Like Peptide-1 on ?- and ?-Cell Function in Isolated Islet and Whole Pancreas Transplant Recipients

    doi: 10.1210/jc.2008-1806

    Figure Lengend Snippet: Plasma glucose levels before and 30 min after GLP-1 7–36 amide ( filled columns ) or placebo ( lined columns ) infusion, and at the end of the approximately 230 mg/dl hyperglycemic clamp. Against the opposite y-axis, M represents the glucose infusion rate required during the hyperglycemic clamp. A, Results for the islet transplant group (n = 5). B, Results for the pancreas transplant group (n = 6). To convert glucose to mmol/liter, multiply by 0.05551.

    Article Snippet: In the islet group, insulin levels were significantly greater after 30 min of GLP-1 compared to placebo infusion (21.6 ± 7.8 vs .

    Techniques:

    Plasma insulin before and after bolus injections of 5 g arginine under both basal and approximately 230 mg/dl hyperglycemic clamp conditions. A, Results for the islet transplant group (n = 5) during GLP-1 7–36 amide (▪) or placebo (□) infusion. B, Results for the pancreas transplant group (n = 6) during GLP-1 7–36 amide (•) or placebo (○) infusion. To convert insulin to pmol/liter, multiply by 7.1750.

    Journal: The Journal of Clinical Endocrinology and Metabolism

    Article Title: Effect of Glucagon-Like Peptide-1 on ?- and ?-Cell Function in Isolated Islet and Whole Pancreas Transplant Recipients

    doi: 10.1210/jc.2008-1806

    Figure Lengend Snippet: Plasma insulin before and after bolus injections of 5 g arginine under both basal and approximately 230 mg/dl hyperglycemic clamp conditions. A, Results for the islet transplant group (n = 5) during GLP-1 7–36 amide (▪) or placebo (□) infusion. B, Results for the pancreas transplant group (n = 6) during GLP-1 7–36 amide (•) or placebo (○) infusion. To convert insulin to pmol/liter, multiply by 7.1750.

    Article Snippet: In the islet group, insulin levels were significantly greater after 30 min of GLP-1 compared to placebo infusion (21.6 ± 7.8 vs .

    Techniques:

    Plasma glucagon before and after bolus injections of 5 g arginine under both basal and approximately 230 mg/dl hyperglycemic clamp conditions. A, Results for the islet transplant group (n = 5) during GLP-1 7–36 amide (▪) or placebo (□) infusion. B, Results for the pancreas transplant group (n = 6) during GLP-1 7–36 amide (•) or placebo (○) infusion.

    Journal: The Journal of Clinical Endocrinology and Metabolism

    Article Title: Effect of Glucagon-Like Peptide-1 on ?- and ?-Cell Function in Isolated Islet and Whole Pancreas Transplant Recipients

    doi: 10.1210/jc.2008-1806

    Figure Lengend Snippet: Plasma glucagon before and after bolus injections of 5 g arginine under both basal and approximately 230 mg/dl hyperglycemic clamp conditions. A, Results for the islet transplant group (n = 5) during GLP-1 7–36 amide (▪) or placebo (□) infusion. B, Results for the pancreas transplant group (n = 6) during GLP-1 7–36 amide (•) or placebo (○) infusion.

    Article Snippet: In the islet group, insulin levels were significantly greater after 30 min of GLP-1 compared to placebo infusion (21.6 ± 7.8 vs .

    Techniques:

    A, Correlation of AIR pot , a measure of β-cell secretory capacity, to the GLP-1 7–36 amide induced change in second-phase insulin levels in both islet (○) and pancreas (•) transplant recipients. B, Correlation of the percentage change in second-phase insulin levels to the percentage change in the glucose infusion rate required during the hyperglycemic clamp in both islet (○) and pancreas (•) transplant recipients. To convert insulin to pmol/liter, multiply by 7.1750.

    Journal: The Journal of Clinical Endocrinology and Metabolism

    Article Title: Effect of Glucagon-Like Peptide-1 on ?- and ?-Cell Function in Isolated Islet and Whole Pancreas Transplant Recipients

    doi: 10.1210/jc.2008-1806

    Figure Lengend Snippet: A, Correlation of AIR pot , a measure of β-cell secretory capacity, to the GLP-1 7–36 amide induced change in second-phase insulin levels in both islet (○) and pancreas (•) transplant recipients. B, Correlation of the percentage change in second-phase insulin levels to the percentage change in the glucose infusion rate required during the hyperglycemic clamp in both islet (○) and pancreas (•) transplant recipients. To convert insulin to pmol/liter, multiply by 7.1750.

    Article Snippet: In the islet group, insulin levels were significantly greater after 30 min of GLP-1 compared to placebo infusion (21.6 ± 7.8 vs .

    Techniques:

    Plasma levels of active GLP-1 7–36 amide in the islet transplant group (n = 5) during GLP-1 7–36 amide (▪) or placebo (□) infusion, and in the pancreas transplant group (n = 6) during GLP-1 7–36 amide (•) or placebo (○) infusion.

    Journal: The Journal of Clinical Endocrinology and Metabolism

    Article Title: Effect of Glucagon-Like Peptide-1 on ?- and ?-Cell Function in Isolated Islet and Whole Pancreas Transplant Recipients

    doi: 10.1210/jc.2008-1806

    Figure Lengend Snippet: Plasma levels of active GLP-1 7–36 amide in the islet transplant group (n = 5) during GLP-1 7–36 amide (▪) or placebo (□) infusion, and in the pancreas transplant group (n = 6) during GLP-1 7–36 amide (•) or placebo (○) infusion.

    Article Snippet: In the islet group, insulin levels were significantly greater after 30 min of GLP-1 compared to placebo infusion (21.6 ± 7.8 vs .

    Techniques:

    GLP-1 potentiates exocytosis in B-cells. Increases in cell capacitance (ΔC m , bottom trace) elicited by progressively longer (5, 10, 15, 30, 50, 100, 150, 250, 350, 450, and 850 ms) depolarizations from −70 to 0 mV (V, top trace) under (A) control conditions and (B) 4 min after the inclusion of 10 nM GLP-1 in the extracellular medium. The interval between two successive depolarizations was 15 s for pulses ≤50 ms and 30 s for longer pulses. For clarity, only responses to the 30-, 100-, 250-, and 450-ms depolarizations are shown. The recording was performed using the perforated patch configuration. (C) Relationship between pulse duration ( t ) and increase in cell capacitance (ΔC m ) under control conditions and in the presence of 10 nM GLP-1 as indicated. The curves were derived by fitting Eq. 3 to the data points for depolarizations ≤350 ms. The dotted lines represent linear fits to the values measured in response to the two longest depolarizations. Data are mean values ± SEM of seven paired experiments. *P

    Journal: The Journal of General Physiology

    Article Title: SUR1 Regulates PKA-independent cAMP-induced Granule Priming in Mouse Pancreatic B-cells

    doi: 10.1085/jgp.20028707

    Figure Lengend Snippet: GLP-1 potentiates exocytosis in B-cells. Increases in cell capacitance (ΔC m , bottom trace) elicited by progressively longer (5, 10, 15, 30, 50, 100, 150, 250, 350, 450, and 850 ms) depolarizations from −70 to 0 mV (V, top trace) under (A) control conditions and (B) 4 min after the inclusion of 10 nM GLP-1 in the extracellular medium. The interval between two successive depolarizations was 15 s for pulses ≤50 ms and 30 s for longer pulses. For clarity, only responses to the 30-, 100-, 250-, and 450-ms depolarizations are shown. The recording was performed using the perforated patch configuration. (C) Relationship between pulse duration ( t ) and increase in cell capacitance (ΔC m ) under control conditions and in the presence of 10 nM GLP-1 as indicated. The curves were derived by fitting Eq. 3 to the data points for depolarizations ≤350 ms. The dotted lines represent linear fits to the values measured in response to the two longest depolarizations. Data are mean values ± SEM of seven paired experiments. *P

    Article Snippet: The effects of GLP-1 were dose-dependent and bell-shaped in both wild-type and SUR1−/− islets.

    Techniques: Mass Spectrometry, Derivative Assay

    GLP-1–induced cAMP production in wild-type and SUR1 −/− B-cells. The amount of cAMP generated was measured in wild-type and SUR1 −/− B-cells as indicated at 0–100 nM GLP-1. Values observed in the presence of 10 μM forskolin have been included for comparison. Data are mean values ± SEM of six experiments in each group. All values in the presence of GLP-1 are statistically different from control value (P

    Journal: The Journal of General Physiology

    Article Title: SUR1 Regulates PKA-independent cAMP-induced Granule Priming in Mouse Pancreatic B-cells

    doi: 10.1085/jgp.20028707

    Figure Lengend Snippet: GLP-1–induced cAMP production in wild-type and SUR1 −/− B-cells. The amount of cAMP generated was measured in wild-type and SUR1 −/− B-cells as indicated at 0–100 nM GLP-1. Values observed in the presence of 10 μM forskolin have been included for comparison. Data are mean values ± SEM of six experiments in each group. All values in the presence of GLP-1 are statistically different from control value (P

    Article Snippet: The effects of GLP-1 were dose-dependent and bell-shaped in both wild-type and SUR1−/− islets.

    Techniques: Generated

    MVR presented as the mean plateau value of the acoustic intensity (AI) in vastus lateralis muscle at basal and after 5-, 10-, and 60-min infusion ( A ) of GLP-1 in the locally infused leg, ( B ) infusion of GLP-1 with coinfusion of octreotide in the locally

    Journal: American Journal of Physiology - Endocrinology and Metabolism

    Article Title: GLP-1 increases microvascular recruitment but not glucose uptake in human and rat skeletal muscle

    doi: 10.1152/ajpendo.00283.2013

    Figure Lengend Snippet: MVR presented as the mean plateau value of the acoustic intensity (AI) in vastus lateralis muscle at basal and after 5-, 10-, and 60-min infusion ( A ) of GLP-1 in the locally infused leg, ( B ) infusion of GLP-1 with coinfusion of octreotide in the locally

    Article Snippet: GLP-1 (7–36 amide; Polypeptide Laboratories, Hillerød, DK) was diluted into 25 ml saline + 5 ml human albumin (5%) and administered as a constant femoral arterial infusion at 1 pmol·kg−1 ·min−1 for 60 min. Due to the expected insulinotropic effect of GLP-1, a slow, arm vein glucose (20% Fresenius Kabi, Upsala, SE) infusion (∼0.5–1 mg·kg−1 ·min−1 ) was initiated after 15 min of GLP-1 infusion to prevent hypoglycemia.

    Techniques:

    Schematic outline of the experiments. denotes sampling of arterialized and femoral venous blood. A : timeline for the GLP-1 and saline (negative control) experiments. B, top : timeline for the GLP-1 infusion with a pre-/coinfusion of octreotide. B, bottom

    Journal: American Journal of Physiology - Endocrinology and Metabolism

    Article Title: GLP-1 increases microvascular recruitment but not glucose uptake in human and rat skeletal muscle

    doi: 10.1152/ajpendo.00283.2013

    Figure Lengend Snippet: Schematic outline of the experiments. denotes sampling of arterialized and femoral venous blood. A : timeline for the GLP-1 and saline (negative control) experiments. B, top : timeline for the GLP-1 infusion with a pre-/coinfusion of octreotide. B, bottom

    Article Snippet: GLP-1 (7–36 amide; Polypeptide Laboratories, Hillerød, DK) was diluted into 25 ml saline + 5 ml human albumin (5%) and administered as a constant femoral arterial infusion at 1 pmol·kg−1 ·min−1 for 60 min. Due to the expected insulinotropic effect of GLP-1, a slow, arm vein glucose (20% Fresenius Kabi, Upsala, SE) infusion (∼0.5–1 mg·kg−1 ·min−1 ) was initiated after 15 min of GLP-1 infusion to prevent hypoglycemia.

    Techniques: Sampling, Negative Control

    A : MVR presented as the mean plateau value of the AI in rat hindlimb muscles at basal and after 75-min infusion of saline or saline + GLP-1. B : 2-deoxyglucose uptake in mixed gastrocnemius-soleus muscle after saline or saline + GLP-1 infusion. Different

    Journal: American Journal of Physiology - Endocrinology and Metabolism

    Article Title: GLP-1 increases microvascular recruitment but not glucose uptake in human and rat skeletal muscle

    doi: 10.1152/ajpendo.00283.2013

    Figure Lengend Snippet: A : MVR presented as the mean plateau value of the AI in rat hindlimb muscles at basal and after 75-min infusion of saline or saline + GLP-1. B : 2-deoxyglucose uptake in mixed gastrocnemius-soleus muscle after saline or saline + GLP-1 infusion. Different

    Article Snippet: GLP-1 (7–36 amide; Polypeptide Laboratories, Hillerød, DK) was diluted into 25 ml saline + 5 ml human albumin (5%) and administered as a constant femoral arterial infusion at 1 pmol·kg−1 ·min−1 for 60 min. Due to the expected insulinotropic effect of GLP-1, a slow, arm vein glucose (20% Fresenius Kabi, Upsala, SE) infusion (∼0.5–1 mg·kg−1 ·min−1 ) was initiated after 15 min of GLP-1 infusion to prevent hypoglycemia.

    Techniques:

    GLP-1R-mediated Ca 2+  signaling in HEK-GLP-1R cells. HEK-GLP-1R cells or wild-type HEK-293 cells were grown in ELISA 8-well strips (96-well format) and used either with or without loading with the Ca 2+  indicator, fluo-4 as indicated.  A ) Fluo-4-loaded HEK-GLP-1R cells were challenged with a range of concentrations of GLP-1 7–36 amide and fluorescence recorded as an index of [Ca 2+ ] i . Fluorescence was calibrated to [Ca 2+ ] i  as described to enable determination of agonist potency. The pEC 50  for the peak of GLP-1 7–36 amide-mediated elevations of [Ca 2+ ] i  was 9.61±0.25 (n = 3).  B ) Fluo-4-loaded HEK-GLP-1R cells were challenged with either buffer, GLP-1 7–36 amide (10 nM) or GLP-1 9–36 amide (10 µM). Alternatively, cells were pretreated with thapsigargin (2 µM, 5 min) to deplete intracellular Ca 2+  stores before challenge with GLP-1 9–36 amide (10 µM). Responses to 30 µM GLP-1 9–36 amide were similar to that evoked by 10 µM (data not shown).  C ) Fluo-4-loaded wild-type HEK-293 cells (i.e. cells without expression of the GLP-1R) were challenged with compound 2 (100 µM) in the absence or presence of pretreatment with thapsigargin (2 µM, 5 min).  D ) Fluo-4-loaded HEK-GLP-1R cells were challenged with either vehicle control (1% DMSO) or compound 2 (100 µM) in the absence or presence of pretreatment with thapsigargin (2 µM, 5 min).  E ) Fluo-4-loaded HEK-GLP-1R cells were challenged with vehicle control (1% DMSO), a concentration of either compound 2 (10 µM) or GLP-1 9–36 amide (1 µM) established to have little effect on [Ca 2+ ] i  or alternatively, compound 2 (10 µM) and GLP-1 9–36 amide (1 µM) in combination. Additionally, cells were challenged with either compound 2 (10 µM) or compound 2 (10 µM) and GLP-1 9–36 amide (1 µM) in combination following pretreatment with thapsigargin (2 µM, 5 min). DMSO was present in all conditions. All data are representative of 3 independent experiments showing similar results.

    Journal: PLoS ONE

    Article Title: Allosteric Modulation of the Activity of the Glucagon-like Peptide-1 (GLP-1) Metabolite GLP-1 9-36 Amide at the GLP-1 Receptor

    doi: 10.1371/journal.pone.0047936

    Figure Lengend Snippet: GLP-1R-mediated Ca 2+ signaling in HEK-GLP-1R cells. HEK-GLP-1R cells or wild-type HEK-293 cells were grown in ELISA 8-well strips (96-well format) and used either with or without loading with the Ca 2+ indicator, fluo-4 as indicated. A ) Fluo-4-loaded HEK-GLP-1R cells were challenged with a range of concentrations of GLP-1 7–36 amide and fluorescence recorded as an index of [Ca 2+ ] i . Fluorescence was calibrated to [Ca 2+ ] i as described to enable determination of agonist potency. The pEC 50 for the peak of GLP-1 7–36 amide-mediated elevations of [Ca 2+ ] i was 9.61±0.25 (n = 3). B ) Fluo-4-loaded HEK-GLP-1R cells were challenged with either buffer, GLP-1 7–36 amide (10 nM) or GLP-1 9–36 amide (10 µM). Alternatively, cells were pretreated with thapsigargin (2 µM, 5 min) to deplete intracellular Ca 2+ stores before challenge with GLP-1 9–36 amide (10 µM). Responses to 30 µM GLP-1 9–36 amide were similar to that evoked by 10 µM (data not shown). C ) Fluo-4-loaded wild-type HEK-293 cells (i.e. cells without expression of the GLP-1R) were challenged with compound 2 (100 µM) in the absence or presence of pretreatment with thapsigargin (2 µM, 5 min). D ) Fluo-4-loaded HEK-GLP-1R cells were challenged with either vehicle control (1% DMSO) or compound 2 (100 µM) in the absence or presence of pretreatment with thapsigargin (2 µM, 5 min). E ) Fluo-4-loaded HEK-GLP-1R cells were challenged with vehicle control (1% DMSO), a concentration of either compound 2 (10 µM) or GLP-1 9–36 amide (1 µM) established to have little effect on [Ca 2+ ] i or alternatively, compound 2 (10 µM) and GLP-1 9–36 amide (1 µM) in combination. Additionally, cells were challenged with either compound 2 (10 µM) or compound 2 (10 µM) and GLP-1 9–36 amide (1 µM) in combination following pretreatment with thapsigargin (2 µM, 5 min). DMSO was present in all conditions. All data are representative of 3 independent experiments showing similar results.

    Article Snippet: Such dependency on both receptor expression levels and the concentration of competing ligand (intact GLP-1), coupled with the possibility of an additional receptor, highlight the difficulties in assessing physiological roles of GLP-1 9–36 amide and may account for some of the discrepancies in the literature.

    Techniques: Enzyme-linked Immunosorbent Assay, Fluorescence, Expressing, Concentration Assay

    Compound 2 potentiates GLP-1 9–36 amide-mediated cAMP generation in INS-1E cells. Cells were stimulated for 15 min with either GLP-1 9–36 amide or compound 2 at the concentrations indicated. Alternatively, cells were stimulated with increasing concentrations of GLP-1 9–36 amide as indicated in the presence of 3 µM compound 2. Responses to GLP-1 7–36 amide are shown for comparison. Data are mean±s.e.m., n = 3.

    Journal: PLoS ONE

    Article Title: Allosteric Modulation of the Activity of the Glucagon-like Peptide-1 (GLP-1) Metabolite GLP-1 9-36 Amide at the GLP-1 Receptor

    doi: 10.1371/journal.pone.0047936

    Figure Lengend Snippet: Compound 2 potentiates GLP-1 9–36 amide-mediated cAMP generation in INS-1E cells. Cells were stimulated for 15 min with either GLP-1 9–36 amide or compound 2 at the concentrations indicated. Alternatively, cells were stimulated with increasing concentrations of GLP-1 9–36 amide as indicated in the presence of 3 µM compound 2. Responses to GLP-1 7–36 amide are shown for comparison. Data are mean±s.e.m., n = 3.

    Article Snippet: Such dependency on both receptor expression levels and the concentration of competing ligand (intact GLP-1), coupled with the possibility of an additional receptor, highlight the difficulties in assessing physiological roles of GLP-1 9–36 amide and may account for some of the discrepancies in the literature.

    Techniques:

    Functional interaction of compound 2 and GLP-1 9–36 amide at the GLP-1R in HEK-GLP-1R cells. A ) Cells were stimulated for 15 min with GLP-1 9–36 amide at the concentrations indicated and either vehicle (KHB+5% DMSO) or compound 2 at the indicated concentrations in the presence of IBMX. The pEC 50  values for GLP-1 9–36 amide-mediated cAMP generation in the presence of the different concentrations of compound 2 are given in   Table 1 .  B ) Data are shown from panel ‘a’ but with the subtraction of the response to compound 2 in the absence of GLP-1 9–36 amide.  C ) The E max  values of cAMP generation at 1 µM GLP-1 9–36 amide in the presence of various concentrations of compound 2 (Co-addition) are compared to the numerical addition of responses to the agonists individually (Numerical addition). Data are taken from panel a.  D ) HEK-GLP-1R cells were stimulated with either isoproterenol (100 µM), compound 2 (1 µM) or the two in combination (Co-addition) in the presence of IBMX and 5% DMSO (vehicle). The numerical sum of cAMP generation in response to isoproterenol and compound 2 is shown (Numerical addition). All data are mean±/+s.e.m., n = 3–4, *** P

    Journal: PLoS ONE

    Article Title: Allosteric Modulation of the Activity of the Glucagon-like Peptide-1 (GLP-1) Metabolite GLP-1 9-36 Amide at the GLP-1 Receptor

    doi: 10.1371/journal.pone.0047936

    Figure Lengend Snippet: Functional interaction of compound 2 and GLP-1 9–36 amide at the GLP-1R in HEK-GLP-1R cells. A ) Cells were stimulated for 15 min with GLP-1 9–36 amide at the concentrations indicated and either vehicle (KHB+5% DMSO) or compound 2 at the indicated concentrations in the presence of IBMX. The pEC 50 values for GLP-1 9–36 amide-mediated cAMP generation in the presence of the different concentrations of compound 2 are given in Table 1 . B ) Data are shown from panel ‘a’ but with the subtraction of the response to compound 2 in the absence of GLP-1 9–36 amide. C ) The E max values of cAMP generation at 1 µM GLP-1 9–36 amide in the presence of various concentrations of compound 2 (Co-addition) are compared to the numerical addition of responses to the agonists individually (Numerical addition). Data are taken from panel a. D ) HEK-GLP-1R cells were stimulated with either isoproterenol (100 µM), compound 2 (1 µM) or the two in combination (Co-addition) in the presence of IBMX and 5% DMSO (vehicle). The numerical sum of cAMP generation in response to isoproterenol and compound 2 is shown (Numerical addition). All data are mean±/+s.e.m., n = 3–4, *** P

    Article Snippet: Such dependency on both receptor expression levels and the concentration of competing ligand (intact GLP-1), coupled with the possibility of an additional receptor, highlight the difficulties in assessing physiological roles of GLP-1 9–36 amide and may account for some of the discrepancies in the literature.

    Techniques: Functional Assay

    Time course of cAMP generation in response to GLP-1 9–36 amide, compound 2 or co-stimulation in HEK-GLP-1R cells. HEK-GLP-1R cells were either untreated (Basal; not visible) or treated for the indicated times with GLP-1 9–36 amide (1 µM), compound 2 (1 µM) or the two in combination (Co-addition) in the presence of IBMX. The final concentration of DMSO (vehicle) was 5% v/v in all cases. In addition to the measured levels of cAMP generation, the numerical sum of cAMP generation in response to GLP-1 9–36 amide and compound 2 alone are presented (Numerical). Data are mean±s.e.m., n = 3, ** P

    Journal: PLoS ONE

    Article Title: Allosteric Modulation of the Activity of the Glucagon-like Peptide-1 (GLP-1) Metabolite GLP-1 9-36 Amide at the GLP-1 Receptor

    doi: 10.1371/journal.pone.0047936

    Figure Lengend Snippet: Time course of cAMP generation in response to GLP-1 9–36 amide, compound 2 or co-stimulation in HEK-GLP-1R cells. HEK-GLP-1R cells were either untreated (Basal; not visible) or treated for the indicated times with GLP-1 9–36 amide (1 µM), compound 2 (1 µM) or the two in combination (Co-addition) in the presence of IBMX. The final concentration of DMSO (vehicle) was 5% v/v in all cases. In addition to the measured levels of cAMP generation, the numerical sum of cAMP generation in response to GLP-1 9–36 amide and compound 2 alone are presented (Numerical). Data are mean±s.e.m., n = 3, ** P

    Article Snippet: Such dependency on both receptor expression levels and the concentration of competing ligand (intact GLP-1), coupled with the possibility of an additional receptor, highlight the difficulties in assessing physiological roles of GLP-1 9–36 amide and may account for some of the discrepancies in the literature.

    Techniques: Concentration Assay

    Compound 2 potentiates GLP-1 9–36 amide-mediated cAMP generation by membranes from HEK-GLP-1R cells. Cell membranes prepared from HEK-GLP-1R cells were incubated with ligands as indicated for 5 min at 30°C in the presence of IBMX before determination of cAMP. A) Concentration-dependent cAMP generation in response to GLP-1 7-36 amide or to GLP-1 9–36 amide either alone or in combination with 3 µM compound 2.  B ) Responses to GLP-1 9–36 amide or compound 2 alone or the two in combination. The numerical sums of cAMP generation in response to GLP-1 9–36 amide and compound 2 are shown. Data are mean±/+s.e.m., n = 3. For * P

    Journal: PLoS ONE

    Article Title: Allosteric Modulation of the Activity of the Glucagon-like Peptide-1 (GLP-1) Metabolite GLP-1 9-36 Amide at the GLP-1 Receptor

    doi: 10.1371/journal.pone.0047936

    Figure Lengend Snippet: Compound 2 potentiates GLP-1 9–36 amide-mediated cAMP generation by membranes from HEK-GLP-1R cells. Cell membranes prepared from HEK-GLP-1R cells were incubated with ligands as indicated for 5 min at 30°C in the presence of IBMX before determination of cAMP. A) Concentration-dependent cAMP generation in response to GLP-1 7-36 amide or to GLP-1 9–36 amide either alone or in combination with 3 µM compound 2. B ) Responses to GLP-1 9–36 amide or compound 2 alone or the two in combination. The numerical sums of cAMP generation in response to GLP-1 9–36 amide and compound 2 are shown. Data are mean±/+s.e.m., n = 3. For * P

    Article Snippet: Such dependency on both receptor expression levels and the concentration of competing ligand (intact GLP-1), coupled with the possibility of an additional receptor, highlight the difficulties in assessing physiological roles of GLP-1 9–36 amide and may account for some of the discrepancies in the literature.

    Techniques: Incubation, Concentration Assay

    ERK signaling through the GLP-1R. HEK-GLP-1R cells ( A and B ) or INS-1E cells ( C and D ) were challenged for 5 min with either vehicle (1% v/v DMSO), GLP-1 9–36 amide (1 or 10 µM as indicated) or compound 2 alone or GLP-1 9–36 amide (1 µM) and compound 2 together as indicated. Cells were also challenged with GLP-1 7–36 amide (10 nM). Levels of phospho-ERK were then determined by Western blotting. The intensity of the bands representing phospho-ERK was determined using ImageJ and the mean data are shown in the panels below the immunoblot with basal (0) levels subtracted. Data are either representative of 3 experiments or mean+s.e.m., n = 3. *, P

    Journal: PLoS ONE

    Article Title: Allosteric Modulation of the Activity of the Glucagon-like Peptide-1 (GLP-1) Metabolite GLP-1 9-36 Amide at the GLP-1 Receptor

    doi: 10.1371/journal.pone.0047936

    Figure Lengend Snippet: ERK signaling through the GLP-1R. HEK-GLP-1R cells ( A and B ) or INS-1E cells ( C and D ) were challenged for 5 min with either vehicle (1% v/v DMSO), GLP-1 9–36 amide (1 or 10 µM as indicated) or compound 2 alone or GLP-1 9–36 amide (1 µM) and compound 2 together as indicated. Cells were also challenged with GLP-1 7–36 amide (10 nM). Levels of phospho-ERK were then determined by Western blotting. The intensity of the bands representing phospho-ERK was determined using ImageJ and the mean data are shown in the panels below the immunoblot with basal (0) levels subtracted. Data are either representative of 3 experiments or mean+s.e.m., n = 3. *, P

    Article Snippet: Such dependency on both receptor expression levels and the concentration of competing ligand (intact GLP-1), coupled with the possibility of an additional receptor, highlight the difficulties in assessing physiological roles of GLP-1 9–36 amide and may account for some of the discrepancies in the literature.

    Techniques: Western Blot

    Functional interaction between ligands on GLP-1R-mediated cAMP generation in HEK-GLP-1R cells. HEK-GLP-1R cells were pretreated (Pre-) for 10 min with 1 µM GLP-1 9–36 amide in the presence of IBMX before challenge for 15 min with the indicated concentrations of agonists. Where no pre-treatment is indicated, an equivalent volume of buffer (KHB) was added for 10 min in the presence of IBMX prior to ligand addition for 15 min. Levels of intracellular cAMP were then determined relative to the cellular protein content. The final concentration of DMSO (vehicle) for the 15 min treatment period was 5% v/v in all cases. Data are mean±s.e.m., n = 3.

    Journal: PLoS ONE

    Article Title: Allosteric Modulation of the Activity of the Glucagon-like Peptide-1 (GLP-1) Metabolite GLP-1 9-36 Amide at the GLP-1 Receptor

    doi: 10.1371/journal.pone.0047936

    Figure Lengend Snippet: Functional interaction between ligands on GLP-1R-mediated cAMP generation in HEK-GLP-1R cells. HEK-GLP-1R cells were pretreated (Pre-) for 10 min with 1 µM GLP-1 9–36 amide in the presence of IBMX before challenge for 15 min with the indicated concentrations of agonists. Where no pre-treatment is indicated, an equivalent volume of buffer (KHB) was added for 10 min in the presence of IBMX prior to ligand addition for 15 min. Levels of intracellular cAMP were then determined relative to the cellular protein content. The final concentration of DMSO (vehicle) for the 15 min treatment period was 5% v/v in all cases. Data are mean±s.e.m., n = 3.

    Article Snippet: Such dependency on both receptor expression levels and the concentration of competing ligand (intact GLP-1), coupled with the possibility of an additional receptor, highlight the difficulties in assessing physiological roles of GLP-1 9–36 amide and may account for some of the discrepancies in the literature.

    Techniques: Functional Assay, Concentration Assay

    Amino acid sequence of GLP-1 and exendin-4. GLP-1 consists of two active circulating forms, GLP-1 (7–36) amide and GLP-1 (7–37).

    Journal: ISRN Endocrinology

    Article Title: Polymer-Based Delivery of Glucagon-Like Peptide-1 for the Treatment of Diabetes

    doi: 10.5402/2012/340632

    Figure Lengend Snippet: Amino acid sequence of GLP-1 and exendin-4. GLP-1 consists of two active circulating forms, GLP-1 (7–36) amide and GLP-1 (7–37).

    Article Snippet: Sustained Release by Encapsulation of GLP-1 or Exendin-4 (1) Controlled Release of GLP-1 Peptide Using Triblock Copolymer of PLGA-PEG-PLGA (ReGel) Long-term delivery of active GLP-1 is required for the treatment of diabetes by peptide delivery.

    Techniques: Sequencing

    GLP-1R silencing abrogates GLP-1-induced signalling pathway activation. ARPE-19 cells were transfected with scrambled (scr) siRNA control or GLP-1R siRNA and treated with 10 nmol/L GLP-1 for 10 minutes. Cells were then lysed and tested with antibody anti-GLP-1R, anti-phPKB, or anti-phERK1/2. Membranes were stripped and reprobed, respectively, with anti- β actin, anti-PKB, or antiERK1/2-antibodies. (a) Representative western blot analyses of three different experiments are shown. (b) Quantification of densitometries of western blot bands of GLP-1R. Data were expressed as mean ± SE of fold induction relative to β -actin ( n = 3). *** P

    Journal: Mediators of Inflammation

    Article Title: Retinal Pigment Epithelial Cells Express a Functional Receptor for Glucagon-Like Peptide-1 (GLP-1)

    doi: 10.1155/2013/975032

    Figure Lengend Snippet: GLP-1R silencing abrogates GLP-1-induced signalling pathway activation. ARPE-19 cells were transfected with scrambled (scr) siRNA control or GLP-1R siRNA and treated with 10 nmol/L GLP-1 for 10 minutes. Cells were then lysed and tested with antibody anti-GLP-1R, anti-phPKB, or anti-phERK1/2. Membranes were stripped and reprobed, respectively, with anti- β actin, anti-PKB, or antiERK1/2-antibodies. (a) Representative western blot analyses of three different experiments are shown. (b) Quantification of densitometries of western blot bands of GLP-1R. Data were expressed as mean ± SE of fold induction relative to β -actin ( n = 3). *** P

    Article Snippet: To evaluate intracellular signalling pathways potentially activated by GLP-1, cells were cultured in serum-free medium for 24 hours and in the absence or presence of 10 nmol/L GLP-1 for different time points (up to 40 minutes).

    Techniques: Activation Assay, Transfection, Western Blot

    Pharmacological inhibition of PI3K abrogates GLP-1-induced activation of PKB. ARPE-19 cells were cultured in serum-free medium for 24 hours and stimulated for 10 minutes with 10 nmol/L GLP-1 in the presence or absence of MEK1/2 inhibitor U0126 (10 μ mol/l) or PI3K inhibitor LY294002 (50 μ mol/l), or EGFR inhibitor AG 1478 (0.25 μ mol/l). (a) Representative western blot analysis of PKB phosphorylation (phPKB) and total protein is shown. (b) Quantification of densitometries of western blot bands. Data were expressed as mean ± SE of fold induction relative to total PKB ( n = 3). ** P

    Journal: Mediators of Inflammation

    Article Title: Retinal Pigment Epithelial Cells Express a Functional Receptor for Glucagon-Like Peptide-1 (GLP-1)

    doi: 10.1155/2013/975032

    Figure Lengend Snippet: Pharmacological inhibition of PI3K abrogates GLP-1-induced activation of PKB. ARPE-19 cells were cultured in serum-free medium for 24 hours and stimulated for 10 minutes with 10 nmol/L GLP-1 in the presence or absence of MEK1/2 inhibitor U0126 (10 μ mol/l) or PI3K inhibitor LY294002 (50 μ mol/l), or EGFR inhibitor AG 1478 (0.25 μ mol/l). (a) Representative western blot analysis of PKB phosphorylation (phPKB) and total protein is shown. (b) Quantification of densitometries of western blot bands. Data were expressed as mean ± SE of fold induction relative to total PKB ( n = 3). ** P

    Article Snippet: To evaluate intracellular signalling pathways potentially activated by GLP-1, cells were cultured in serum-free medium for 24 hours and in the absence or presence of 10 nmol/L GLP-1 for different time points (up to 40 minutes).

    Techniques: Inhibition, Activation Assay, Cell Culture, Western Blot

    GLP-1 does not affect ARPE-19 cell viability. Cells were cultured for 24, 72, or 96 hours in the absence (CTR, white bars) or presence of 10 nmol/L GLP-1 (GPL-1, black bars). Cell proliferation rate was determined by a colorimetric method based on the formazan product of the tetrazolium compound MTS. Results showed the percentage of absorbance compared to CTR. Data were expressed as the mean ± SE of 3 independent experiments.

    Journal: Mediators of Inflammation

    Article Title: Retinal Pigment Epithelial Cells Express a Functional Receptor for Glucagon-Like Peptide-1 (GLP-1)

    doi: 10.1155/2013/975032

    Figure Lengend Snippet: GLP-1 does not affect ARPE-19 cell viability. Cells were cultured for 24, 72, or 96 hours in the absence (CTR, white bars) or presence of 10 nmol/L GLP-1 (GPL-1, black bars). Cell proliferation rate was determined by a colorimetric method based on the formazan product of the tetrazolium compound MTS. Results showed the percentage of absorbance compared to CTR. Data were expressed as the mean ± SE of 3 independent experiments.

    Article Snippet: To evaluate intracellular signalling pathways potentially activated by GLP-1, cells were cultured in serum-free medium for 24 hours and in the absence or presence of 10 nmol/L GLP-1 for different time points (up to 40 minutes).

    Techniques: Cell Culture

    GLP-1 does not affect secretion of VEGF-A. ARPE-19 cells were cultured for 24 h in in the presence or absence of 10 nmol/L GLP-1. The conditioned media were collected, and VEGF-A levels in supernatants were assessed by ELISA. VEGF-A concentration was calculated from standards curve and normalized to total protein concentration of the respective lysates. The results are representative of three independent experiments (mean ± SE).

    Journal: Mediators of Inflammation

    Article Title: Retinal Pigment Epithelial Cells Express a Functional Receptor for Glucagon-Like Peptide-1 (GLP-1)

    doi: 10.1155/2013/975032

    Figure Lengend Snippet: GLP-1 does not affect secretion of VEGF-A. ARPE-19 cells were cultured for 24 h in in the presence or absence of 10 nmol/L GLP-1. The conditioned media were collected, and VEGF-A levels in supernatants were assessed by ELISA. VEGF-A concentration was calculated from standards curve and normalized to total protein concentration of the respective lysates. The results are representative of three independent experiments (mean ± SE).

    Article Snippet: To evaluate intracellular signalling pathways potentially activated by GLP-1, cells were cultured in serum-free medium for 24 hours and in the absence or presence of 10 nmol/L GLP-1 for different time points (up to 40 minutes).

    Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Concentration Assay, Protein Concentration

    GLP-1 phosphorylates the intracellular signalling proteins PKB and ERK1/2. ARPE-19 cells were cultured in serum-free medium for 24 hours and stimulated with control medium (0′) or for 10, 20, and 40 minutes with 10 nmol/L GLP-1. Then, cells were then lysed and tested for phosphorylation and total protein of PKB (a) and ERK1/2 (b). Upper panel: representative western blot analysis of three different experiments is shown. Lower panel: densitometries of western blot bands. Data were expressed as mean ± SE of fold induction relative to total protein ( n = 3). ** P

    Journal: Mediators of Inflammation

    Article Title: Retinal Pigment Epithelial Cells Express a Functional Receptor for Glucagon-Like Peptide-1 (GLP-1)

    doi: 10.1155/2013/975032

    Figure Lengend Snippet: GLP-1 phosphorylates the intracellular signalling proteins PKB and ERK1/2. ARPE-19 cells were cultured in serum-free medium for 24 hours and stimulated with control medium (0′) or for 10, 20, and 40 minutes with 10 nmol/L GLP-1. Then, cells were then lysed and tested for phosphorylation and total protein of PKB (a) and ERK1/2 (b). Upper panel: representative western blot analysis of three different experiments is shown. Lower panel: densitometries of western blot bands. Data were expressed as mean ± SE of fold induction relative to total protein ( n = 3). ** P

    Article Snippet: To evaluate intracellular signalling pathways potentially activated by GLP-1, cells were cultured in serum-free medium for 24 hours and in the absence or presence of 10 nmol/L GLP-1 for different time points (up to 40 minutes).

    Techniques: Cell Culture, Western Blot

    Pharmacological inhibition of PI3K blocked GLP-1-induced activation of ERK1/2. ARPE-19 cells were cultured in serum-free medium for 24 hours and stimulated for 10 minutes with 10 nmol/L GLP-1 in the presence or absence of MEK1/2 inhibitor U0126 (10 μ mol/l) or PI3K inhibitor LY294002 (50 μ mol/l), or EGFR inhibitor AG 1478 (0.25 μ mol/l). (a) Representative western blot analysis of ERK 1/2 phosphorylation (phERK1/2) and total protein is shown. (b) Quantification of densitometries of western blot bands of phERK1. Data were expressed as mean ± SE of fold induction relative to total ERK1 ( n = 3). ** P

    Journal: Mediators of Inflammation

    Article Title: Retinal Pigment Epithelial Cells Express a Functional Receptor for Glucagon-Like Peptide-1 (GLP-1)

    doi: 10.1155/2013/975032

    Figure Lengend Snippet: Pharmacological inhibition of PI3K blocked GLP-1-induced activation of ERK1/2. ARPE-19 cells were cultured in serum-free medium for 24 hours and stimulated for 10 minutes with 10 nmol/L GLP-1 in the presence or absence of MEK1/2 inhibitor U0126 (10 μ mol/l) or PI3K inhibitor LY294002 (50 μ mol/l), or EGFR inhibitor AG 1478 (0.25 μ mol/l). (a) Representative western blot analysis of ERK 1/2 phosphorylation (phERK1/2) and total protein is shown. (b) Quantification of densitometries of western blot bands of phERK1. Data were expressed as mean ± SE of fold induction relative to total ERK1 ( n = 3). ** P

    Article Snippet: To evaluate intracellular signalling pathways potentially activated by GLP-1, cells were cultured in serum-free medium for 24 hours and in the absence or presence of 10 nmol/L GLP-1 for different time points (up to 40 minutes).

    Techniques: Inhibition, Activation Assay, Cell Culture, Western Blot