ampk activator aicar  (MedChemExpress)

Journal: Journal of Advanced Research

Article Title: Prophylactic supplementation with biogenic selenium nanoparticles mitigated intestinal barrier oxidative damage through suppressing epithelial-immune crosstalk with gut-on-a-chip

doi: 10.1016/j.jare.2025.04.023

Figure Lengend Snippet: Biogenic SeNPs regulated AMPK/NLRP3/Nrf2 signaling pathway to alleviate intestinal epithelial barrier oxidative damage in gut-on-a-chip. A. Schematic diagram of experimental design. B. pAMPK/NLRP3/Nrf2 immunofluorescent staining (Scale bar: 40 μm). C. Villi-like height. D. LDH activity in the upper channel layer. E. IL-1β levels in the upper channel layer. F. IL-18 levels in the upper channel layer. G. Schematic diagram of the mechanism by which SeNPs exert antioxidant effects against oxidative damage to intestinal epithelial cells in vitro . Data are expressed as mean ± S.E.M. n = 4. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: To investigate the role of the AMPK signaling pathway in the protection of the intestinal epithelial barrier from oxidative stress damage by SeNPs, AMPK activator AICAR (MedChemExpres; Cat# HY-13417) and AMPK inhibitor Dorsomorphin (MedChemExpres; Cat# HY-13418A) were introduced into the gut-on-a-chip, respectively.

Techniques: Staining, Activity Assay, In Vitro

Journal: Journal of Advanced Research

Article Title: Prophylactic supplementation with biogenic selenium nanoparticles mitigated intestinal barrier oxidative damage through suppressing epithelial-immune crosstalk with gut-on-a-chip

doi: 10.1016/j.jare.2025.04.023

Figure Lengend Snippet: Biogenic SeNPs regulated AMPK/NLRP3/Nrf2 signaling pathway to attenuate oxidative stress-induced intestinal barrier dysfunction and mast cell overactivation in mice. A. Immunofluorescence analysis of p-AMPK (red), NLRP3 (yellow) and Nrf2 (green) in jejunum of mice with different treatments (Scale bar: 100 μm). B. Western blot analysis of pAMPK and Nrf2 expression levels in mice jejunum. C. Western blot analysis of NLRP3 and its downstream pyroptosis-related protein expression levels in mice jejunum. Data are expressed as mean ± S.E.M. n = 3. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: To investigate the role of the AMPK signaling pathway in the protection of the intestinal epithelial barrier from oxidative stress damage by SeNPs, AMPK activator AICAR (MedChemExpres; Cat# HY-13417) and AMPK inhibitor Dorsomorphin (MedChemExpres; Cat# HY-13418A) were introduced into the gut-on-a-chip, respectively.

Techniques: Immunofluorescence, Western Blot, Expressing

Journal: iScience

Article Title: Roseburia hominis enriched by baicalin reverses the non-response to metformin via upregulating linolenic acid metabolism

doi: 10.1016/j.isci.2025.113892

Figure Lengend Snippet: Baicalin combined with metformin improved the blood glucose levels in metformin non-responsive mice by activating the AMPK/ACC/CPT1 pathway (A–F) The serum concentrations of (A) TC, (B) LDL-C, (C) HDL-C, (D) IL-1β, (E) IL-6, and (F) IL-10. (G) Hematoxylin-eosin (H&E) staining of the liver, pancreas, epididymal adipose tissues, and oil red O staining of the liver (magnification, 30×; scale bars, 100 μm). (H) Western blot analysis of pAMPK (Thr172), ACC, and CPT1 proteins in the livers of mice after administration. Data are expressed as the mean ± SD (A‒F, n = 6–8; H, n = 3); ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, as determined by one-way ANOVA with Holm-Sidak’s post hoc test (A, B, C, E, F, and H) and Kruskal-Wallis test (D). NCD, normal chow diet; NR, non-response; Met, metformin; BA, baicalin; TC, total cholesterol; LDL-C, low-density lipoprotein cholesterol; HDL-C, high-density lipoprotein cholesterol; IL-1β, interleukin-1β; IL-6, interleukin-6; IL-10, interleukin-10; AMPK, AMP-activated protein kinase; ACC, acetyl-CoA carboxylase; CPT1, carnitine palmitoyl transferase 1.

Article Snippet: These membranes were then incubated at 4°C overnight with primary antibodies against AMP-activated protein kinase (AMPK) (Proteintech, Cat# 10929-2-AP, RRID: AB_2169568 , 1:4,000), insulin receptor substrate 1 (IRS1) (Proteintech, Cat# 17509-1-AP, RRID: AB_10596914 , 1:1,000), phospho-AMPKα (Thr172) (40H9) (Cell Signaling Technology, Cat# 2535, RRID: AB_331250 , 1:1,000), phospho-IRS-1 (Ser636/639) (Cell Signaling Technology, Cat# 2388, RRID: AB_330339 , 1:1,000), phosphoenolpyruvate carboxykinase (PCK) (Proteintech, Cat# 16754-1-AP, RRID: AB_2160031 , 1:30,000), glucose-6-phosphatase (G6PC) (Proteintech, Cat# 66860-1-Ig, RRID: AB_2882199 , 1:5,000), mammalian target of rapamycin (mTOR) (Proteintech, Cat# 66888-1-Ig, RRID: AB_2882219 , 1:30,000), fatty acid desaturase 2 (FADS2) (Proteintech, Cat# 28034-1-AP, RRID: AB_2918142 , 1:4,000), phospholipase A2 group (PLA2G) (Proteintech, Cat# 18088-1-AP, RRID: AB_10859777 , 1:1,000), acetyl-CoA Carboxylase 1 (ACC1) (Proteintech, Cat# 21923-1-AP, RRID: AB_11042445 , 1:4,000), carnitine palmitoyl transferase 1 (CPT1) (Proteintech, Cat# 15184-1-AP, RRID: AB_2084676 , 1:50,000), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Proteintech, Cat# 10494-1-AP, RRID: AB_2263076 , 1:20,000), β-tubulin (Proteintech, Cat# 10094-1-AP, RRID: AB_2210695 , 1:10,000).

Techniques: Staining, Western Blot

Journal: iScience

Article Title: Roseburia hominis enriched by baicalin reverses the non-response to metformin via upregulating linolenic acid metabolism

doi: 10.1016/j.isci.2025.113892

Figure Lengend Snippet: R. hominis treatment reversed the metformin NR phenotype in NR mice (A) Experimental protocol for administration of R. hominis in mice. (B) Oral glucose tolerance test (OGTT) curve and its area under the curve (AUC). (C) Insulin tolerance test (ITT) curve and its AUC. (D) Homeostasis model assessment of insulin resistance (HOMA-IR) after drug R. hominis . (E–J) The serum concentrations of (E) TC, (F) LDL-C, (G) HDL-C, (H) IL-1β, (I) IL-6, and (J) IL-10. (K) Hematoxylin-eosin (H&E) staining of the liver, pancreas, epididymal adipose tissues, and oil red O staining of the liver (magnification, 30×; scale bars, 100 μm). (L) Western blot analysis of AMPK/ACC/CPT1 proteins in the livers of mice after administration. Data are expressed as the mean ± SD ( n = 7–8); ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, as determined by one-way ANOVA with Holm-Sidak’s post hoc test. ANOVA, analysis of variance; ABX, antibiotic mixed; FMT, fecal microbial transplantation; NCD, normal chow diet; R.h, Roseburia hominis ; TC, total cholesterol; LDL-C, low-density lipoprotein cholesterol; HDL-C, high-density lipoprotein cholesterol; IL-1β, interleukin-1β; IL-6, interleukin-6; IL-10, interleukin-10; AMPK, AMP-activated protein kinase; ACC, acetyl-CoA carboxylase; CPT1, carnitine palmitoyl transferase 1.

Article Snippet: These membranes were then incubated at 4°C overnight with primary antibodies against AMP-activated protein kinase (AMPK) (Proteintech, Cat# 10929-2-AP, RRID: AB_2169568 , 1:4,000), insulin receptor substrate 1 (IRS1) (Proteintech, Cat# 17509-1-AP, RRID: AB_10596914 , 1:1,000), phospho-AMPKα (Thr172) (40H9) (Cell Signaling Technology, Cat# 2535, RRID: AB_331250 , 1:1,000), phospho-IRS-1 (Ser636/639) (Cell Signaling Technology, Cat# 2388, RRID: AB_330339 , 1:1,000), phosphoenolpyruvate carboxykinase (PCK) (Proteintech, Cat# 16754-1-AP, RRID: AB_2160031 , 1:30,000), glucose-6-phosphatase (G6PC) (Proteintech, Cat# 66860-1-Ig, RRID: AB_2882199 , 1:5,000), mammalian target of rapamycin (mTOR) (Proteintech, Cat# 66888-1-Ig, RRID: AB_2882219 , 1:30,000), fatty acid desaturase 2 (FADS2) (Proteintech, Cat# 28034-1-AP, RRID: AB_2918142 , 1:4,000), phospholipase A2 group (PLA2G) (Proteintech, Cat# 18088-1-AP, RRID: AB_10859777 , 1:1,000), acetyl-CoA Carboxylase 1 (ACC1) (Proteintech, Cat# 21923-1-AP, RRID: AB_11042445 , 1:4,000), carnitine palmitoyl transferase 1 (CPT1) (Proteintech, Cat# 15184-1-AP, RRID: AB_2084676 , 1:50,000), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Proteintech, Cat# 10494-1-AP, RRID: AB_2263076 , 1:20,000), β-tubulin (Proteintech, Cat# 10094-1-AP, RRID: AB_2210695 , 1:10,000).

Techniques: Staining, Western Blot, Transplantation Assay

Journal: Europace

Article Title: Promoting differentiation of human-induced pluripotent stem cells into sinoatrial node-like cells through programmed regulation of AMPK signalling pathway

doi: 10.1093/europace/euaf288

Figure Lengend Snippet: The role of AMPK signalling pathway in NKX2.5 expression. ( A ) Pan-cardiomyocyte differentiation protocol. CHIR (Wnt agonist) and IWP2 (Wnt inhibitor) were used for directed differentiation of cardiac cells. ( B–D ) Expression trends of markers at various stages during cardiomyocyte differentiation including cardiac mesoderm marker MESP1, cardiomyocyte progenitor marker NKX2.5, and cardiomyocyte marker TNNT2. qRT–PCR analysis was conducted to observe the dynamic expression trends from Days 0 to 13 of induced differentiation. ( E–F ) KEGG analysis of signalling pathways enriched with NXK2.5 expression. ( G ) Different concentrations of the AMPK agonist AICAR (AA) were added on Day 4 of differentiation for 2 days. The expression level of NKX2.5 in each group was analysed by qRT–PCR on Day 6. ( H ) Different concentrations of AMPK inhibitor CC were added on Day 4 of differentiation for 2 days. The expression level of NKX2.5 in each group was analysed by qRT–PCR on Day 6. One-way ANOVA followed by Bonferroni post hoc test was performed, * P < 0.05, *** P < 0.001, **** P < 0.0001, and ns, not significant vs. control, N = 5. All PCR expression values were normalized to the housekeeping gene GAPDH. Data are presented as ‘mean ± SEM’.

Article Snippet: Following the cardiomyocyte differentiation protocol outlined above, varying concentrations of AMPK activator AICAR (AA) (0, 100, 200, 500, 800, 1000 μM, HY-13417, MedChemExpress) were added individually between Days 4 and 6 of differentiation.

Techniques: Expressing, Marker, Quantitative RT-PCR, Control

Journal: Europace

Article Title: Promoting differentiation of human-induced pluripotent stem cells into sinoatrial node-like cells through programmed regulation of AMPK signalling pathway

doi: 10.1093/europace/euaf288

Figure Lengend Snippet: Activation of the AMPK signalling pathway on the differentiation of hiPSCs to SANLCs. ( A ) Schematic diagram of the experimental protocol. ( B ) Different concentrations of an AMPK AA were added on Day 4 of differentiation for 2 days, and the expression levels of SAN cell markers SHOX2 , TBX18 , TBX3 , HCN4 , and pan-cardiomyocyte marker TNNT2 were analysed by qRT–PCR on Day 21 of differentiation. ( C ) AA was added on Days 4–6, 6–8, and 4–8, respectively, and the expression levels of SAN cell markers SHOX2 , TBX18 , TBX3 , and HCN4 and pan-cardiomyocyte marker TNNT2 were analysed by qRT–PCR on Day 21 of differentiation. One-way ANOVA followed by Bonferroni post hoc test was performed (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns, not significant vs. control, N = 5). All PCR expression values were normalized to the housekeeping gene GAPDH. Data are presented as ‘mean ± SEM’.

Article Snippet: Following the cardiomyocyte differentiation protocol outlined above, varying concentrations of AMPK activator AICAR (AA) (0, 100, 200, 500, 800, 1000 μM, HY-13417, MedChemExpress) were added individually between Days 4 and 6 of differentiation.

Techniques: Activation Assay, Expressing, Marker, Quantitative RT-PCR, Control

Journal: Europace

Article Title: Promoting differentiation of human-induced pluripotent stem cells into sinoatrial node-like cells through programmed regulation of AMPK signalling pathway

doi: 10.1093/europace/euaf288

Figure Lengend Snippet: Inhibition of the AMPK signalling pathway on the differentiation of hiPSCs to SANLCs. ( A ) Schematic diagram of the experimental protocol. ( B ) Different concentrations of an AMPK inhibitor CC were added on Day 6 of differentiation induction for 2 days, and the expression levels of SAN cell markers SHOX2 , TBX18 , TBX3 , and HCN4 and pan-cardiomyocyte marker TNNT2 were analysed by qRT–PCR on Day 21 of differentiation. ( C ) AA was added on Days 6–8, 8–10, and 6–10, and the expression levels of SHOX2, TBX18, TBX3 , HCN4 , and TNNT2 were analysed by qRT–PCR on Day 21 of differentiation. One-way ANOVA followed by Bonferroni post hoc test was performed (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns, not significant vs. control, N = 5). All PCR expression values were normalized to the housekeeping gene GAPDH. Data are presented as ‘mean ± SEM’.

Article Snippet: Following the cardiomyocyte differentiation protocol outlined above, varying concentrations of AMPK activator AICAR (AA) (0, 100, 200, 500, 800, 1000 μM, HY-13417, MedChemExpress) were added individually between Days 4 and 6 of differentiation.

Techniques: Inhibition, Expressing, Marker, Quantitative RT-PCR, Control