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、100μmol/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100μmol/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100μmol/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100μmol/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、100μmol/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。
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(25,50,100μmol/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 100μmol/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 100μmol/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 100μmol/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 100μmol/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.
作者
王瑞瑶
霍梦露
李超
关锡梅
倪伟建
唐丽琴
Wang Ruiyao;Huo Menglu;Li Chao;Guan Ximei;Ni Weijian;Tang Liqin(School of Pharmacy,Anhui University of Chinese Medicine,Hefei 230031,China)
出处
《中国临床保健杂志》
CAS
2020年第2期241-246,共6页
Chinese Journal of Clinical Healthcare
基金
国家自然科学基金项目(81773955)
安徽省中医药科研项目(2016zy08)。
作者简介
王瑞瑶,硕士研究生,Email:masterchief00@163.com;通讯作者:唐丽琴,教授,博士研究生导师,Email:tangliqin@ustc.edu.cn。
