抑制JWA基因表达对细胞因子TGF-β1诱导的心肌细胞凋亡的机制研究  

Study on the Mechanism of Inhibitory Effect of JWA Gene Expression on Cardiac Myocyte Apoptosis Induced by TGF-β1

作　　者：胡国平[1] HU Guo-ping(Hubei Polytechnic Institute,Xiaogan, Hubei 432000, China)

机构地区：[1]湖北职业技术学院,湖北孝感432000

出　　处：《湖北职业技术学院学报》2020年第3期93-99,62,共8页Journal of Hubei Polytechnic Institute

摘　　要：目的:研究抑制JWA基因的表达对转化生长因子-β1(TGF-β1)诱导的心肌细胞凋亡的影响及作用机制。方法:用TGF-β1转染JWA RNA处理体外培养大鼠心肌H9c2心肌细胞,采用反转录聚合酶链技术(RT-PCR)和Western blot法检测心肌细胞中JWA的表达水平的变化,采用噻唑蓝(MTT)检测心肌细胞增殖情况,流式细胞术检测心肌细胞凋亡情况,对比检测心肌细胞中凋亡相关蛋白剪切的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved Caspase-3)蛋白、Bcl-2相关X(Bax)蛋白和Bcl-2蛋白水平,检测心肌细胞中产生的活性氧(ROS)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-PX)和丙二醛(MDA)的含量。结果:TGF-β1处理后的心肌细胞中JWA mRNA水平和Bcl-2蛋白水平、CAT、GSH-PX含量均明显高于正常培养的心肌细胞,细胞存活率升高,细胞凋亡率和细胞中Cleaved Caspase-3、Bax蛋白水平、Bax/Bcl-2值、细胞中ROS、MDA含量均降低(P<0.05),与正常培养的心肌细胞相比,差异有统计学意义。结论:抑制JWA的表达可减少TGF-β1诱导的心肌细胞凋亡,促进心肌细胞增殖,减弱TGF-β1诱导的心肌细胞氧化损伤,其作用机制可能是通过减轻细胞的氧化损伤减少细胞凋亡。Objective:To study the effect of inhibiting the expression of JWA gene on the apoptosis of cardiomyocytes induced by transforming growth factor-β1(TGF-β1)and its mechanism of action.Methods:Myocardial H9c2 cells were cultured in vitro and cardiomyocytes were treated with TGF-β1.Reverse transcriptase polymerase chain reaction(RT-PCR)and western blot were used to detect the expression of JWA in cardiomyocytes.The proliferation of cardiomyocytes was detected by thiazolyl blue(MTT).The apoptosis of cardiomyocytes was detected by flow cytometry.Comparison was made to determine the cystein-induced cleavage of apoptosis-related proteins in cardiomyocytes.Caspase-3(Cleaved Caspase-3)protein,Bcl-2 related X(Bax)protein,and Bcl-2 protein levels detect superoxide dismutase(SOD)produced in cardiomyocytes,Catalase(CAT),glutathione peroxidase(GSH-PX)and malondialdehyde(MDA)content.Results:The level of JWA mRNA and protein in cardiomyocytes after TGF-β1 treatment was significantly higher than that of normal cultured cardiomyocytes(P<0.05).The survival rate of cardiomyocytes after TGF-β1 treatment increased,and the rate of apoptosis,Cleaved Caspase-3 and Bax protein levels in the cells and Bcl-2 protein levels decreased.The content of ROS and MDA in the cells decreased.Compared with normal cultured cardiomyocytes,the difference was statistically significant(P<0.05).Conclusion:Inhibiting the expression of JWA may reduce the apoptosis of cardiomyocytes induced by TGF-β1,promote the proliferation of cardiomyocytes and reduce the oxidative damage induced by TGF-β1 in cardiomyocytes.The mechanism may reduce the oxidative damage of cells and reduce the apoptosis.

关 键 词：JWA 心肌细胞 TGF-Β1 凋亡 机制研究 

分 类 号：R542.2[医药卫生—心血管疾病]