Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Reduced NGF in Gastric Endothelial Cells Is One of the Main Causes of Impaired Angiogenesis in Aging Gastric Mucosa

doi: 10.1016/j.jcmgh.2018.05.003

Figure Lengend Snippet: NGF treatment reverses impairment of angiogenesis in aging GECs through PI3 kinase/Akt and mTOR pathways. ( A ) NGF treatment dose-dependently increases in vitro angiogenesis in aging GECs. Aging GECs were treated with either PBS (control) or exogenous NGF (10, 50, 100, or 1000 ng/mL) and in vitro angiogenesis on Matrigel was examined. Data are means ± SD (N = 6). Treatment of aging GECs with NGF dose-dependently increased angiogenesis with the maximal threshold stimulating dose of 100 ng/mL at 6 hours ( P < .001). There was no significant difference in angiogenesis between cells treated with NGF 100 ng/mL and 1000 ng/mL. † P > .05 (NS). ( B ) In vitro angiogenesis on Matrigel at 6 hours in aging GECs treated with PBS or recombinant rat NGF (100 ng/mL) with or without pretreatment with specific PI3 kinase inhibitor (LY294002; 50 μmol/L for 30 minutes), or mTOR inhibitor (rapamycin; 10 nmol/L for 60 minutes). Short-term (6 hours) treatment with exogenous NGF significantly increased in vitro angiogenesis and in aging GECs by 1.5-fold and pretreatment with LY294002, and rapamycin abolished the NGF-induced increase in angiogenesis. Data are means ± SD (N = 6). ( C ) Representative Western blots for phosphorylated Akt (pAkt) and Akt expression in aging GECs treated with exogenous NGF (0.1 μg/mL) for 4 and 24 hours. Treatment of aging GECs with exogenous NGF significantly increased Akt phosphorylation at 4 and 24 hours. Data are means ± SD (N = 3). The Student t test and 1-way analysis of variance with the Tukey multiple comparison test were used to determine statistical significance between 2 or multiple groups, respectively.

Article Snippet: For some studies, GECs were treated with the specific PI3 kinase inhibitor 50 μmol/L for 30 minutes; Cell Signaling Technology, Danvers, MA), the mTOR inhibitor rapamycin (10 nmol/L for 60 minutes; Cell Signaling Technology), Latrunculin B (0.5 μmol/L for 30 minutes; Invitrogen, Carlsbad, CA; inhibitor of actin polymerization), or recombinant rat NGF (10–1000 ng/mL; R&D Systems, Minneapolis, MN).

Techniques: In Vitro, Recombinant, Western Blot, Expressing

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Curcumin Alleviates D-Galactose-Induced Cardiomyocyte Senescence by Promoting Autophagy via the SIRT1/AMPK/mTOR Pathway

doi: 10.1155/2022/2990843

Figure Lengend Snippet: Curcumin enhanced the SIRT1-AMPK/mTOR signal pathway. (a) Representative western blot for SIRT1 p-AMPK and p-mTOR levels in cardiomyocyte lysates ( n = 5). (b) Quantitation results of p-mTOR. (c) Quantitation results of SIRT1. (d). Quantitation results of p-AMPK. ∗∗ P < 0.01 vs. control. # P < 0.05, # # P < 0.01, vs. D-gal. The data are expressed as the mean ± SEM.

Article Snippet: The membranes were washed three times with TBST and incubated with a secondary antibody labeled with horseradish peroxidase (1 : 5000; Abclonal, China) at room temperature for 1 h. The primary rabbit antibodies included mTOR (1 : 1000; Cell Signaling Technology, USA), phospho-mTOR (1 : 1000; Cell Signaling Technology, USA), AMPK (1 : 1000; Cell Signaling Technology, USA), phospho-AMPK (1 : 1000; Cell Signaling Technology, USA), SIRT1 (1 : 1000; Cell Signaling Technology, USA), P53 (1 : 500; Abclonal, China, A19585), P16 (1 : 2000; Abcam, USA), LC3I/II (1 : 2000; Abcam, USA), P62 (1 : 1000; Cell Signaling Technology, USA), and GAPDH (1 : 1000; Cell Signaling Technology, USA).

Techniques: Western Blot, Quantitation Assay

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Curcumin Alleviates D-Galactose-Induced Cardiomyocyte Senescence by Promoting Autophagy via the SIRT1/AMPK/mTOR Pathway

doi: 10.1155/2022/2990843

Figure Lengend Snippet: (a) Representative western blot of the levels of SIRT1 p-AMPK and p-mTOR. siSIRT1-mediated knockdown blocked the inducing effect of curcumin on the SIRT1/AMPK/mTOR pathway in cardiac aging ( n = 5). (b) The quantitation results of SIRT1. (c) The quantitation results of p-AMPK. (d) The quantitation results of p-mTOR. # P < 0.05, # # P < 0.01, vs. control. The data are expressed as the mean ± SEM.

Article Snippet: The membranes were washed three times with TBST and incubated with a secondary antibody labeled with horseradish peroxidase (1 : 5000; Abclonal, China) at room temperature for 1 h. The primary rabbit antibodies included mTOR (1 : 1000; Cell Signaling Technology, USA), phospho-mTOR (1 : 1000; Cell Signaling Technology, USA), AMPK (1 : 1000; Cell Signaling Technology, USA), phospho-AMPK (1 : 1000; Cell Signaling Technology, USA), SIRT1 (1 : 1000; Cell Signaling Technology, USA), P53 (1 : 500; Abclonal, China, A19585), P16 (1 : 2000; Abcam, USA), LC3I/II (1 : 2000; Abcam, USA), P62 (1 : 1000; Cell Signaling Technology, USA), and GAPDH (1 : 1000; Cell Signaling Technology, USA).

Techniques: Western Blot, Quantitation Assay

Journal: Neural Plasticity

Article Title: Growth Hormone Increases BDNF and mTOR Expression in Specific Brain Regions after Photothrombotic Stroke in Mice

doi: 10.1155/2022/9983042

Figure Lengend Snippet: List of antibodies used for western blot analyses.

Article Snippet: T-mTOR , Cell Signalling, monoclonal mouse anti-total- mTOR, #4517 , 1 : 1000.

Techniques: Western Blot

Journal: Neural Plasticity

Article Title: Growth Hormone Increases BDNF and mTOR Expression in Specific Brain Regions after Photothrombotic Stroke in Mice

doi: 10.1155/2022/9983042

Figure Lengend Snippet: t -test and Pearson's results summary table.

Article Snippet: T-mTOR , Cell Signalling, monoclonal mouse anti-total- mTOR, #4517 , 1 : 1000.

Techniques:

Journal: Neural Plasticity

Article Title: Growth Hormone Increases BDNF and mTOR Expression in Specific Brain Regions after Photothrombotic Stroke in Mice

doi: 10.1155/2022/9983042

Figure Lengend Snippet: The effect of recombinant human growth factor (r-hGH) treatment poststroke on mammalian target of rapamycin (mTOR) in the peri-infarct region. (a) Representative blots of phosphorylated-mTOR (P-mTOR, active form), total-mTOR (T-mTOR), phosphorylated-p70S6-kinase P-p70S6K), and total-p70S6k (T-p70S6K). (b) T-mTOR and T-p70S6K levels were normalized to β -actin. P-mTOR and P-p70S6K levels were normalized to T-mTOR and T-p70S6K, respectively. Levels were expressed as a fold change (FC) of mean ± SD for each group relative to the mean of the Sham+Saline group (dotted line). We found a significant increase in T-mTOR protein expression within the peri-infarct region in stroked mice treated with r-hGH (blue squares) compared with saline (red circles). ∗ p < 0.05, 2-tailed t -test.

Article Snippet: T-mTOR , Cell Signalling, monoclonal mouse anti-total- mTOR, #4517 , 1 : 1000.

Techniques: Recombinant, Expressing

Journal: Neural Plasticity

Article Title: Growth Hormone Increases BDNF and mTOR Expression in Specific Brain Regions after Photothrombotic Stroke in Mice

doi: 10.1155/2022/9983042

Figure Lengend Snippet: The effect of recombinant human growth factor (r-hGH) treatment poststroke on mammalian target of rapamycin (mTOR) in the hippocampus. (a) Representative blots of phosphorylated-mTOR (P-mTOR, active form), total-mTOR (T-mTOR), phosphorylated-p70s6-kinase (P-p70S6K), and total-p70s6k (T-p70S6K). (b) Data are presented as in . We found a significant increase in T-p70S6K protein expression within the hippocampus in stroked mice treated with r-hGH (blue squares) compared with saline (red circles). ∗ p < 0.05, 2-tailed t -test.

Article Snippet: T-mTOR , Cell Signalling, monoclonal mouse anti-total- mTOR, #4517 , 1 : 1000.

Techniques: Recombinant, Expressing

Journal: Neural Plasticity

Article Title: Growth Hormone Increases BDNF and mTOR Expression in Specific Brain Regions after Photothrombotic Stroke in Mice

doi: 10.1155/2022/9983042

Figure Lengend Snippet: The effect of recombinant human growth factor (r-hGH) treatment poststroke on mammalian target of rapamycin (mTOR) in the thalamus. (a) Representative blots of phosphorylated-mTOR (P-mTOR, active form), total-mTOR (T-mTOR), phosphorylated-p70S6-kinase (P-p70S6K), and total-p70S6K (T-p70S6K). (b) Data are presented as in . We found a significant increase in T-mTOR protein expression within the thalamus in stroked mice treated with r-hGH (blue squares) compared with saline (red circles). ∗∗ p < 0.01, 2-tailed t -test.

Article Snippet: T-mTOR , Cell Signalling, monoclonal mouse anti-total- mTOR, #4517 , 1 : 1000.

Techniques: Recombinant, Expressing

Journal: Neural Plasticity

Article Title: Growth Hormone Increases BDNF and mTOR Expression in Specific Brain Regions after Photothrombotic Stroke in Mice

doi: 10.1155/2022/9983042

Figure Lengend Snippet: Correlation of uncleaved immature form of BDNF (pro-BDNF) with mammalian target of rapamycin (mTOR) and markers of neuroplasticity. Correlation of pro-BDNF, with T-mTOR (top panel), α -amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-receptor subunit GluR1 (marker of synaptic plasticity, second from top panel), NeuN (neuronal marker, third from top panel), and doublecortin (DCX, neuronal migration marker, bottom panel) in the peri-infarct region (a), hippocampus (b), and thalamus (c). Dataset GluR1, NeuN, and DCX was an excerpt from previous study [ , ]. Correlations contain all three experimental groups, Sham+Saline (black crosses), Stroke+Saline (red circles), and Stroke+r-hGH (blue squares) and analyzed using Pearson's correlation. Pearson's r and associated p values for each correlation are reported on the corresponding graph.

Article Snippet: T-mTOR , Cell Signalling, monoclonal mouse anti-total- mTOR, #4517 , 1 : 1000.

Techniques: Marker, Migration

Journal: Neural Plasticity

Article Title: Growth Hormone Increases BDNF and mTOR Expression in Specific Brain Regions after Photothrombotic Stroke in Mice

doi: 10.1155/2022/9983042

Figure Lengend Snippet: List of antibodies used for western blot analyses.

Article Snippet: P-mTOR , Cell Signalling, monoclonal rabbit anti-phospho-mTOR (Ser2448), #5536 , 1 : 500.

Techniques: Western Blot

Journal: Neural Plasticity

Article Title: Growth Hormone Increases BDNF and mTOR Expression in Specific Brain Regions after Photothrombotic Stroke in Mice

doi: 10.1155/2022/9983042

Figure Lengend Snippet: t -test and Pearson's results summary table.

Article Snippet: P-mTOR , Cell Signalling, monoclonal rabbit anti-phospho-mTOR (Ser2448), #5536 , 1 : 500.

Techniques:

Journal: Neural Plasticity

Article Title: Growth Hormone Increases BDNF and mTOR Expression in Specific Brain Regions after Photothrombotic Stroke in Mice

doi: 10.1155/2022/9983042

Figure Lengend Snippet: The effect of recombinant human growth factor (r-hGH) treatment poststroke on mammalian target of rapamycin (mTOR) in the peri-infarct region. (a) Representative blots of phosphorylated-mTOR (P-mTOR, active form), total-mTOR (T-mTOR), phosphorylated-p70S6-kinase P-p70S6K), and total-p70S6k (T-p70S6K). (b) T-mTOR and T-p70S6K levels were normalized to β -actin. P-mTOR and P-p70S6K levels were normalized to T-mTOR and T-p70S6K, respectively. Levels were expressed as a fold change (FC) of mean ± SD for each group relative to the mean of the Sham+Saline group (dotted line). We found a significant increase in T-mTOR protein expression within the peri-infarct region in stroked mice treated with r-hGH (blue squares) compared with saline (red circles). ∗ p < 0.05, 2-tailed t -test.

Article Snippet: P-mTOR , Cell Signalling, monoclonal rabbit anti-phospho-mTOR (Ser2448), #5536 , 1 : 500.

Techniques: Recombinant, Expressing

Journal: Neural Plasticity

Article Title: Growth Hormone Increases BDNF and mTOR Expression in Specific Brain Regions after Photothrombotic Stroke in Mice

doi: 10.1155/2022/9983042

Figure Lengend Snippet: The effect of recombinant human growth factor (r-hGH) treatment poststroke on mammalian target of rapamycin (mTOR) in the hippocampus. (a) Representative blots of phosphorylated-mTOR (P-mTOR, active form), total-mTOR (T-mTOR), phosphorylated-p70s6-kinase (P-p70S6K), and total-p70s6k (T-p70S6K). (b) Data are presented as in . We found a significant increase in T-p70S6K protein expression within the hippocampus in stroked mice treated with r-hGH (blue squares) compared with saline (red circles). ∗ p < 0.05, 2-tailed t -test.

Article Snippet: P-mTOR , Cell Signalling, monoclonal rabbit anti-phospho-mTOR (Ser2448), #5536 , 1 : 500.

Techniques: Recombinant, Expressing

Journal: Neural Plasticity

Article Title: Growth Hormone Increases BDNF and mTOR Expression in Specific Brain Regions after Photothrombotic Stroke in Mice

doi: 10.1155/2022/9983042

Figure Lengend Snippet: The effect of recombinant human growth factor (r-hGH) treatment poststroke on mammalian target of rapamycin (mTOR) in the thalamus. (a) Representative blots of phosphorylated-mTOR (P-mTOR, active form), total-mTOR (T-mTOR), phosphorylated-p70S6-kinase (P-p70S6K), and total-p70S6K (T-p70S6K). (b) Data are presented as in . We found a significant increase in T-mTOR protein expression within the thalamus in stroked mice treated with r-hGH (blue squares) compared with saline (red circles). ∗∗ p < 0.01, 2-tailed t -test.

Article Snippet: P-mTOR , Cell Signalling, monoclonal rabbit anti-phospho-mTOR (Ser2448), #5536 , 1 : 500.

Techniques: Recombinant, Expressing