Ginsenoside Rb1 Ameliorates Autophagy of Hypoxia Cardiomyocytes from Neonatal Rats via AMP-Activated Protein Kinase Pathway

• 来源期刊: 中国结合医学杂志（英文版）
• 年，卷（期）: 2019,（7）
• 页码范围: 521-528
• 页数: 8页
• 语种: 英文
• DOI: 10.1007/s11655-018-3018-y

摘要

Objective:To investigate whether ginsenoside-Rb1 (Gs-Rb1) improves the CoCl2-induced autophagy of cardiomyocytes via upregulation of adenosine 5'-monophosphate-activated protein kinase (AMPK) pathway.Methods:Ventricles from 1-to 3-day-old Wistar rats were sequentially digested,separated and incubated in Dulbecco's modified Eagle's medium supplemented with 10％ fetal bovine serum for 3 days followed by synchronization.Neonatal rat cardiomyocytes were randomly divided into 7 groups:control group (normal level oxygen),hypoxia group (500 μ mol/L CoCl2),Gs-Rb1 group (200 μ mol/L Gs-Rb1 + 500 μ mol/L CoCl2),Ara A group (500 μ mol/L Ara A + 500 μ mol/L CoCl2),Ara A+ Gs-Rb1 group (500 μ mol/L Ara A + 200 μ mol/LGs-Rb1 + 500 μ mol/L CoCl2),AICAR group [1 mmol/L 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR) + 500 μ mol/L CoCl2],and AICAR+Gs-Rb1 group (1 mmol/L AICAR + 200 μ mol/L Gs-Rb1 + 500 μ mol/L CoCl2).Cells were treated for 12 h and cell viability was determined by methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay and cardiac troponin I (cTnl) levels were detected by enzyme-linked immunosorbent assay (ELISA).AMPK activity was assessed by 2',7'-dichlorofluorescein diacetate (DCFH-DA) ELISA assay.The protein expressions of Atg4B,Atg5,Atg6,Atg7,microtubule-associated protein 1A/1B-light chain 3 (LC3),P62,and active-cathepsin B were measured by Western blot.Results:Gs-Rb1 significantly improved the cell viability of hypoxia cardiomyocytes (P＜0.01).However,the viability of hypoxia-treated cardiomyocytes was significantly inhibited by Ara A (P＜0.01).Gs-Rb1 increased the AMPK activity of hypoxia-treated cardiomyocytes.The AMPK activity of hypoxia-treated cadiomyocytes was inhibited by Ara A (P＜0.01) and was not affected by AICAR (P=0.983).Gs-Rb1 up-regulated Atg4B,Atg5,Beclin-1,Atg7,LC3B Ⅱ,the LC3BⅡ/Ⅰ ratio and cathepsin B activity of hypoxia cardiomyocytes (P＜0.05),each of these protein levels was significantly enhanced by Ara A (all P＜0.01),but was not affected by AICAR (all P＞0.05).Gs-Rb1 significantly down-regulated P62 levels of hypoxic cardiomyocytes (P＜0.05).The P62 levels of hypoxic cardiomyocytes were inhibited by Ara A (P＜0.05) and were not affected by AICAR (P=0.871).Conclusion:Gs-Rb1 may improve the viability of hypoxia cardiomyocytes by ameliorating cell autophagy via the upregulation of AMPK pathway.