EZH2调节PI3K/Akt/mTOR信号通路对高糖诱导的心肌细胞凋亡和自噬的影响*

doi:10.3870/j.issn.1672-0741.24.12.020

摘要/Abstract

摘要： 目的探讨Zeste增强子同源物2(EZH2)调节磷脂酰肌醇3激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白(PI3K/Akt/mTOR)通路对高糖诱导的心肌细胞凋亡和自噬的影响。方法小鼠心肌细胞(MCM细胞)分为对照组、高糖组、高糖+过表达对照组、高糖+EZH2过表达组、高糖+沉默对照组、高糖+EZH2沉默组、高糖+EZH2沉默+PI3K抑制剂(BKM120)组。qRT-PCR检测MCM细胞EZH2 mRNA表达。CCK-8法检测MCM细胞活力。ELISA法检测MCM细胞上清液中乳酸脱氢酶(LDH)水平。流式细胞术检测MCM细胞凋亡。透射电镜观察细胞内自噬小体的数量。Western blot法检测MCM细胞EZH2、Bax、LC3-Ⅱ/LC3-Ⅰ、p62、p-PI3K、p-Akt、p-mTOR蛋白表达。结果与对照组比较,高糖组MCM细胞EZH2 mRNA表达,细胞上清液LDH水平、细胞凋亡率、自噬小体数量及EZH2、Bax、LC3-Ⅱ/LC3-Ⅰ蛋白升高,MCM细胞活力及p62、p-PI3K、p-Akt、p-mTOR蛋白表达降低(均P<0.05)。与高糖组、高糖+过表达对照组比较,高糖+EZH2过表达组MCM细胞EZH2 mRNA表达、细胞上清液中LDH水平、细胞凋亡率、自噬小体数量及EZH2、Bax、LC3-Ⅱ/LC3-Ⅰ蛋白表达升高,MCM细胞活力及p62、p-PI3K、p-Akt、p-mTOR蛋白表达降低(均P<0.05)。与高糖组、高糖+沉默对照组比较,高糖+EZH2沉默组MCM细胞EZH2 mRNA表达、细胞上清液LDH水平、细胞凋亡率、自噬小体数量及EZH2、Bax、LC3-Ⅱ/LC3-Ⅰ蛋白表达降低,MCM细胞活力及p62、p-PI3K、p-Akt、p-mTOR蛋白表达升高(均P<0.05)。BKM120逆转了沉默EZH2对高糖诱导的MCM细胞凋亡和自噬的抑制作用。结论沉默EZH2可能通过激活PI3K/Akt/mTOR通路抑制高糖诱导的MCM细胞凋亡和自噬。

关键词: Zeste增强子同源物2, 糖尿病心肌病, 高糖, 心肌细胞, 凋亡, 自噬

Abstract: Objective To investigate the effect of enhancer of Zeste homolog 2(EZH2)on high glucose-induced cardiomyocyte apoptosis and autophagy by regulating the phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/mTOR)pathway. Methods Mouse cardiomyocytes (MCM cells) were divided into control group,high glucose group,high glucose+overexpression control group,high glucose+EZH2 overexpression group,high glucose+silencing control group,high glucose+EZH2 silencing group,and high glucose+EZH2 silencing+PI3K inhibitor(BKM120)group.The qRT-PCR was applied to detect EZH2 mRNA expression in MCM cells.CCK-8 was applied to detect the viability of MCM cells.ELISA was applied to detect the level of lactate dehydrogenase(LDH)in the supernatant of MCM cells.Flow cytometry was applied to detect apoptosis in MCM cells.Transmission electron microscopy was applied to observe the number of autophagosomes inside cells.Western blot was applied to detect EZH2,Bax,LC3-Ⅱ/LC3-Ⅰ,p62,p-PI3K,p-Akt,and p-mTOR proteins in MCM cells. Results The expression of EZH2 mRNA in MCM cells,level of LDH in cell supernatant,apoptosis rate,number of autophagosomes,and EZH2,Bax,LC3-Ⅱ/LC3-Ⅰproteins in high glucose group were higher,the viability of MCM cells and the expression of p62,p-PI3K,p-Akt,and p-mTOR proteins were lower than in the control group(all P<0.05).Compared with the high glucose group and the high glucose+overexpression control group,the expression of EZH2 mRNA in MCM cells,level of LDH in cell supernatant,apoptosis rate,number of autophagosomes,and EZH2,Bax,LC3-Ⅱ/LC3-Ⅰ proteins in high glucose+EZH2 overexpression group were increased,the viability of MCM cells and the expression of p62,p-PI3K,p-Akt,and p-mTOR proteins were decreased(all P<0.05).Compared with the high glucose group and the high glucose+silencing control group,the expression of EZH2 mRNA in MCM cells,level of LDH in cell supernatant,apoptosis rate,number of autophagosomes,and EZH2,Bax,LC3-Ⅱ/LC3-Ⅰ proteins in the high glucose+EZH2 silencing group were decreased,the viability of MCM cells and the expression of p62,p-PI3K,p-Akt,and p-mTOR proteins were increased(all P<0.05).BKM120 reversed the inhibitory effects of silencing EZH2 on high glucose-induced apoptosis and autophagy in MCM cells. Conclusion Silencing EZH2 may inhibit high glucose-induced apoptosis and autophagy in MCM cells by activating the PI3K/Akt/mTOR pathway.

Key words: enhancer of Zeste homolog 2, diabetes cardiomyopathy, high glucose, cardiomyocytes, apoptosis, autophagy

中图分类号:

李彦伟, 杨贺然, 李兴江, 胡嘉航, 刘贵波. EZH2调节PI3K/Akt/mTOR信号通路对高糖诱导的心肌细胞凋亡和自噬的影响^*[J]. 华中科技大学学报（医学版）, 2026, 55(3): 363-369.

Li Yanwei, Yang Heran, Li Xingjiang et al. Effect of EZH2 on High Glucose-induced Cardiomyocyte Apoptosis and Autophagy by Regulating the PI3K/Akt/mTOR Signaling Pathway[J]. Acta Medicinae Universitatis Scientiae et Technologiae Huazhong, 2026, 55(3): 363-369.

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EZH2调节PI3K/Akt/mTOR信号通路对高糖诱导的心肌细胞凋亡和自噬的影响^*

Effect of EZH2 on High Glucose-induced Cardiomyocyte Apoptosis and Autophagy by Regulating the PI3K/Akt/mTOR Signaling Pathway

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