葛根素激活AMPK/Akt/GSK-3β信号通路减轻高糖诱导的H9c2心肌细胞肥大作用研究

Study on the effect of puerarin on alleviating high glucose-induced hypertrophy of H9c2 cells via activating AMPK/Akt/GSK-3βsignaling pathway

[目的]探讨葛根素对高糖诱导的H9c2心肌细胞的保护作用及机制。[方法]采用高糖损伤H9c2心肌细胞48 h,建立体外糖尿病心肌损伤细胞模型;罗丹明标记的鬼笔环肽标记细胞骨架,测定细胞表面积;蛋白免疫印迹法(Western Blot)检测细胞中心肌肥大相关蛋白β-肌球蛋白重链(β-MHC)的水平、AMP活化蛋白激酶α(AMPKα)、丝氨酸/苏氨酸激酶(Akt)和糖原合成激酶3β(GSK-3β)蛋白的磷酸化水平、以及1,4,5-三磷酸2型受体(IP3R2)和Sigma-1受体(Sig-1R)的蛋白水平。[结果]葛根素(20μmol/L)能显著抑制高糖诱导的心肌细胞表面积增大;Western Blot分析结果显示,高糖能上调β-MHC蛋白水平、抑制AMPK和Akt磷酸化,葛根素能下调β-MHC、IP3R2蛋白水平同时促进AMPK、Akt和GSK-3β的磷酸化。[结论]葛根素可能通过激活AMPK/Akt/GSK-3β信号通路减轻高糖诱导的心肌肥大。

[Objective]The aim of the present study was to examine the effects of puerarin on H9c2 cells injured by high glucose.[Methods]H9c2 cells were injured by high glucose for 48 h,and diabetic myocardial injury cell model was established in vitro.The cytoskeleton was stained with rhodamine phalloidin,and the cell surface area was determined.Western blot was used to detect the expression ofβ-MHC,phosphorylation of AMPKα,Akt and GSK-3β,and protein levels of IP3R2 and SIG-1R.[Results]Puerarin(20μmol/L)could significantly inhibit the increase of cardiomyocyte surface area induced by high glucose.Western blot analysis showed that high glucose up-regulatedβ-MHC protein levels and inhibited phosphorylation of AMPK and Akt,while puerarin down-regulatedβ-MHC and IP3R2 expression and promoted phosphorylation of AMPK,Akt and GSK-3β.[Conclusion]Puerarin may alleviate hypertrophy of H9C2 cells induced by high glucose via activating AMPK/Akt/GSK-3βsignaling pathway.