文章摘要
王瑞瑶,霍梦露,李超,关锡梅,倪伟建,唐丽琴.小檗碱对高糖诱导心肌细胞AMPK-AS160-GLUT4 通路的调节及保护作用[J].中国临床保健杂志,2020,23(2):241-246.
小檗碱对高糖诱导心肌细胞AMPK-AS160-GLUT4 通路的调节及保护作用
Regulation and protective effect of berberine on hyperglycemic-induced AMPK-AS160-GLUT4 pathway in cardiomyocytes
投稿时间:2019-10-17  
DOI:10.3969/J.issn.1672-6790.2020.02.026
中文关键词: 糖尿病心肌病  肌细胞,心脏  蛋白质丝氨酸苏氨酸激酶  小檗碱  细胞保护  细胞凋亡
英文关键词: Diabetic cardiomyopathies  Myocytes,cardiac  Protein-serine-threonine kinases  Berberine  Cytoprotection  Apoptosis 〖FL
基金项目:国家自然科学基金项目(81773955);安徽省中医药科研项目(2016zy08)
作者单位E-mail
王瑞瑶 安徽中医药大学药学院,合肥 230031 masterchief00@163.com 
霍梦露 安徽中医药大学药学院,合肥 230031  
李超 安徽中医药大学药学院,合肥 230031  
关锡梅 安徽中医药大学药学院,合肥 230031  
倪伟建 中国科学技术大学附属第一医院安徽省立医院药剂科  
唐丽琴 中国科学技术大学附属第一医院安徽省立医院药剂科 tangliqin@ustc.edu.cn 
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中文摘要:
      目的 探讨小檗碱对高糖诱导心肌细胞AMPK-AS160-GLUT4通路调节及保护作用。方法 体外培养大鼠H9c2心肌细胞系,设置组别为健康对照组、高糖模型组、不同浓度小檗碱组(25、50、100 μmol/L)并分别处理。CCK-8法检测细胞增殖与毒性;Annexin V-FITC/PI双染实验观察小檗碱对高糖诱导心肌细胞凋亡的影响;鬼笔环肽染色观察心肌细胞骨架;Western blot法检测小檗碱对高糖诱导心肌细胞腺苷酸活化蛋白激酶(AMPK)、磷酸化糖代谢调节蛋白(P-AS160)、葡萄糖转运蛋白4(GLUT4)、丙酮酸脱氢酶(PDH)蛋白表达的影响。结果 CCK-8检测结果可见高糖诱导心肌细胞的细胞活力100 μmol/L小檗碱组显著高于模型组(P<0.05);Annexin V-FITC/PI双染实验结果显示心肌细胞凋亡率100 μmol/L小檗碱组显著低于模型组(P<0.01);心肌细胞骨架观察结果显示,小檗碱可改善心肌细胞肥大情况。Western blot结果显示AMPK与AS160的磷酸化程度小檗碱组明显高于模型组(P<0.01),丙酮酸脱氢酶(PDH)含量100 μmol/L小檗碱组明显高于模型组(P<0.01),心肌细胞膜上GLUT4含量50、100 μmol/L小檗碱组明显高于模型组(P<0.01)。结论 小檗碱可改善高糖诱导心肌细胞损伤,提高心肌细胞葡萄糖代谢效率,其作用机制可能与调控AMPK-AS160-GLUT4信号通路有关。
英文摘要:
      Objective To explore the regulation and protective effect of berberine on hyperglycemic-induced AMPK-AS160-GLUT4 pathway in cardiomyocytes.Methods Rat H9c2 cardiomyocyte cell line was cultured in vitro,and were divided as normal control group,high glucose model group and different concentrations of berberine group (25,50,100 μmol/L) and treated separately.Cell proliferation and toxicity were detected by cck-8 assay;Annexin V-FITC/PI double staining was used to observe the effect of berberine on apoptosis of high glucose-induced cardiomyocytes;observation of myocardial cytoskeleton by ghost pen ring peptide staining.he effects of berberine on the expression of hyperglyceme-induced cardiomyocyte adenylate activated protein kinase (AMPK),phosphorylated glucose metabolism regulatory protein (p-as160),glucose transporters 4(GLUT4) and pyruvate dehydrogenase (PDH) were determined by Western blot.Results CCK-8 test results showed that the cell viability of cardiomyocytes induced by high glucose in the 100 μmol/Lberberine group was significantly higher than that in the model group(P<0.05).Annexin V-FITC/PI double staining results showed that the apoptosis rate of cardiomyocytes in the 100 μmol/L berberine group was significantly lower than that in the model group (P<0.01).Myocardial cytoskeleton observations show that berberine can improve myocardial cell hypertrophy.Western blot results showed that the phosphorylation degree of AMPK and AS160 in the berberine group was significantly higher than that in the model group(P<0.01).The content of pyruvate dehydrogenase(PDH) in the 100 μmol/Lberberine group was significantly higher than that in the model group(P<0.01).The GLUT4 content in the myocardial cell membrane in the 50 and 100 μmol/Lberberine group was significantly higher than that in the model group(P<0.01).Conclusion Berberine can alleviate myocardial cell injury induced by high glucose and improve the glucose metabolism efficiency of myocardial cells,and its mechanism may be related to the regulation of ampk-as160-glut4 signaling pathway.
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