延胡索乙素对高糖诱导H9c2细胞凋亡和氧化应激的抑制作用  

Inhibitory effect of l-tetrahydropalmatine on apoptosis and oxidative stress of H9c2 cells induced by high glucose

作　　者：蹇明辉 陈文明 张怡 Jian Minghui;Chen Wenming;Zhang yi(Department of Cardiology,Affiliated Hospital of Zunyi Medical University,Zunyi Guizhou 563099,China;Department of Health Toxicology,School of Public Health,Zunyi Medical University,Zunyi Guizhou 563099,China)

机构地区：[1]遵义医科大学附属医院心血管内科,贵州遵义563099 [2]遵义医科大学公共卫生学院卫生毒理学教研室,贵州遵义563099

出　　处：《遵义医科大学学报》2022年第3期317-321,共5页Journal of Zunyi Medical University

基　　金：遵义医科大学附属医院硕士启动基金资助项目(NO:院字[2012]06);遵义市科技支撑计划项目(NO:遵市科合HZ字(2020)227;遵市科合HZ字(2021)51)。

摘　　要：目的探讨延胡索乙素(l-THP)对高糖诱导H9c2细胞凋亡和氧化应激抑制作用。方法以高糖诱导H9c2心肌细胞凋亡为实验模型,将实验分为5组:对照组(Control)、模型组(Model)、l-THP高、中、低3个剂量组(100、50、25μmol/L);用噻唑蓝(MTT)法测定细胞活力,用试剂盒法检测乳酸脱氢酶(LDH)、活性氧簇(ROS)、谷胱甘肽过氧化物酶(GSH-Px)的表达量,流式细胞术检测细胞凋亡率,Western blot印迹法检测定半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)蛋白表达水平;RT-PCR法检测Fas和FasL基因表达水平。结果与Control组相比,Model组H9c2细胞的活力明显降低,l-THP能逆转高糖对心肌H9c2细胞活力降低的作用,并呈浓度依赖性;与Control组相比,Model组H9c2细胞LDH的释放量和ROS的活性均明显增加,GSH-PX的活性显著下降,与Model组相比,l-THP可呈浓度依赖性降低LDH的释放量,降低ROS的活性,增强GSH-PX的活性;与Control组相比,Model组H9c2细胞凋亡率明显升高,Caspase-3的蛋白表达水平明显升高,Fas和FasL基因表达水平明显上升,与Model组相比,l-THP能降低H9c2细胞的凋亡率,降低Caspase-3的蛋白表达水平,下调Fas和FasL基因表达水平。结论l-THP通过调控细胞凋亡和抗氧化作用来保护H9c2细胞免受高糖诱导的损伤,以此提高其生存能力。Objective To study the inhibitory effects of tetrahydropalmatine(l-THP)on high glucose induced apoptosis and oxidative stress of H9c2 cells.Methods The apoptosis of H9c2 myocardial cells induced by high glucose was used as the experimental model,and the experiment was divided into 5 groups:Control group,Model group,and l-THP high-dose,medium-dose and low-dose groups(100,50,25μmol/L).The cell viability was determined by thiazoline-blue method,the expression levels of lactate dehydrogenase(LDH),reactive oxygen species(ROS)and glutathione peroxidase(GSH-Px)were detected by kit method,and the apoptosis rate was detected by flow cytometry.Caspase-3 protein expression was detected by Western blot.Fas and FasL gene expression levels were detected by RT-PCR.Results Compared with the Control group,the activity of H9c2 cells in Model group was significantly decreased,and l-THP could reverse the effect of high glucose on the activity of H9c2 cells in a concentration-dependent manner.Compared with the Control group,the release amount of LDH and ROS activity of H9c2 cells in Model group were significantly increased,while the activity of GSH-Px was significantly decreased.Compared with the Model group,l-THP could reduce the release amount of LDH,decrease the activity of ROS,and enhance the activity of GSH-Px in a concentration-dependent manner.Compared with the Control group,the apoptosis rate of H9c2 cells in the Model group was significantly increased,the protein expression level of Caspase-3 was significantly increased,and the expression level of Fas and FasL genes was significantly increased.Compared with the Model group,l-THP could reduce the apoptosis rate of H9c2 cells,decreased caspase-3 protein expression,Fas and FasL gene expression.Conclusion l-THP protects H9c2 cells from high glucose-induced damage by regulating apoptosis and antioxidant effects,thus improving their viability.

关 键 词：H9C2心肌细胞 延胡索乙素 高糖 细胞凋亡 氧化应激 

分 类 号：R542.2[医药卫生—心血管疾病]