绿原酸通过PI3K/Akt信号通路抑制异丙肾上腺素诱导的心肌肥大

Chlorogenic acid inhibits isoproterenol-induced cardiac hypertrophy through PI3K/Akt signaling pathway

目的探讨绿原酸(CGA)通过磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/Akt)信号通路抑制异丙肾上腺素(ISO)诱导的心肌肥大的作用机制。方法采用ISO诱导H9c2心肌细胞,建立心肌肥大模型。将细胞随机分为5组:空白对照组(Control组)、异丙肾上腺素组(ISO组)、绿原酸处理组(ISO+CGA组)、抑制剂LY294002组(ISO+LY组)、绿原酸处理+LY294002(ISO+CGA+LY组)。检测各实验组H9c2心肌细胞的心肌表面积及存活率;各组心肌细胞凋亡率和活性氧类(ROS)水平;各组细胞中Caspase、Akt、p-Akt、Gsk-3β、p-Gsk-3β蛋白的表达水平。结果与Control组相比,ISO组心肌细胞严重受损,细胞存活率明显降低(P<0. 05);比较ISO+CGA组和ISO组,ISO刺激显著增加心肌细胞表面积(50%),而CGA预处理可防止ISO诱导的细胞肥大。与ISO组相比,ISO+CGA组p-Akt、p-Gsk-3β蛋白表达水平升高(P<0. 05),H9c2心肌细胞存活率明显提高(P<0. 05),细胞凋亡率明显降低(P<0. 05),Caspase蛋白表达降低(P<0. 05)。与ISO+CGA组相比,ISO+LY组细胞存活率明显降低(P<0. 05),细胞凋亡率升高(P<0. 05),p-Akt、p-Gsk-3β蛋白表达水平降低(P<0. 05),Caspase蛋白表达升高(P<0. 05); ISO+CGA+LY组的心肌细胞存活率降低(P<0. 05),细胞凋亡率升高(P<0. 05),p-Akt、pGsk-3β蛋白表达水平降低(P<0. 05),Caspase蛋白表达升高(P<0. 05)。结论 CGA可能是通过PI3K/Akt信号通路调控细胞凋亡,进而抑制ISO诱导的心肌细胞肥大。

Objective To investigate the effect of chlorogenic acid(CGA) on isoproterenol(ISO)-induced cardiac hypertrophy and its relation to phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt) signaling pathway. Methods ISO-induced H9c2 myocardial cells were used as a model of myocardial hypertrophy.Cells were randomly divided into 5 groups:blank control group(control group),isoproterenol group(ISO group),chlorogenic acid treatment group(ISO+CGA group),inhibitor LY294002 group(ISO+LY group) and chlorogenic acid treatment +LY294002(ISO+CGA+LY group).The surface area of H9c2 myocardial cells and the survival rate of myocardial cells in each group were determined by the enzyme labeling apparatus.Cell apoptosis and reactive oxygen species(ROS) were measured by flow cytometry.Western blot was used to detect the expression levels of Caspase,Akt,p-Akt,Gsk-3β and p-Gsk-3β proteins in each group. Results Compared with the blank control group,the myocardial cells in the ISO group were severely damaged and the cell survival rate was significantly reduced( P <0.05).ISO stimulation significantly increased the myocardial cell surface area(50%),and the CGA pretreatment prevented iso-induced hypertrophy.Compared with the ISO group,the expression levels of p-Akt and p-Gsk-3β proteins in ISO + CGA group were increased( P <0.05),the survival rate of H9c2 cells was significantly increased( P <0.05),the apoptosis rate was significantly decreased( P <0.05),and the expression of Caspase was decreased( P <0.05).Compared with the ISO+ CGA group,the cell survival rate was significantly reduced( P <0.05),apoptosis rate was increased( P <0.05),p-Akt,p-Gsk-3β protein expression level was decreased( P <0.05),and Caspase protein expression was increased( P <0.05) in ISO+LY group and ISO+CGA+LY group. Conclusion CGA attenuates ISO-induced cardiac hypertrophy,which may be associated with its roles in regulation of apoptosis through the PI3K/Akt signaling pathway.