摘要
目的构建体外高糖诱导的H9c2心肌细胞损伤模型,观察高糖环境下心肌细胞损伤情况及TRAP1的表达变化及意义。方法将体外培养的H9c2细胞随机分为正常糖对照组(5.5 mmol/L葡萄糖)、高糖组(33.0 mmol/L葡萄糖)、高渗组(5.5 mmol/L葡萄糖+27.5 mmol/L甘露醇),体外培养48 h;Western blot及实时荧光定量PCR(qRT-PCR)方法检测各组细胞TRAP1蛋白及mRNA相对表达量;MTS检测心肌细胞活性;2',7'-二氢二氯荧光黄双乙酸盐(DCFH-DA)荧光探针检测细胞内ROS含量;JC-1荧光染色检测心肌细胞线粒体膜电位变化。结果与正常糖对照组及高渗组相比,高糖组H9c2细胞中TRAP1 mRNA及蛋白表达量均下降(P <0.05);细胞活性下降(P <0.05);细胞内ROS含量增多(P <0.05);心肌细胞线粒体膜电位下降(P <0.05)。结论高糖环境下心肌细胞中TRAP1表达的下降,可能与ROS介导的线粒体功能受损所致的心肌损伤密切相关。
Objective To construct the model of myocardial injury induced by high glucose in vitro,and to investigate the expression and function of tumor necrosis factor receptor-associated protein 1(TRAP1)in cardiomyocytes under hyperglycemia.Methods H9c2 cells were treated with normal glucose(5.5 mmol/L Glucose),high glucose(33.0 mmol/L Glucose),and high osmotic pressure(5.5 mmol/L Glucose+ 27.5mmol/L Mannitol)for 48 h.TRAP1 mRNA expression was analyzed by qPCR and TRAP1 protein level was determined by Western blot analysis.Activity of cardiomyocytes was detected using the CellTiter 96? AQueous One Solution assay.The intracellular reactive oxygen species(ROS)content was detected using the fluorescent 2',7'-dichlorodihydrofluorescein diacetate probe,and the change in mitochondrial membrane potential was detected by JC-1 fluorescent staining.Results Compared with normal control and high osmotic groups,the expression of TRAP1 in H9c2 cells decreased(P<0.05)in high glucose group,accompanied by reduction in mitochondrial membrane potential and cell activity,and increased intracellular ROS production(all P<0.05).Conclusions Decreased TRAP1 expression in cardiomyocytes under high glucose conditions is closely related to myocardial injury induced by ROS-mediated mitochondrial dysfunction.
作者
钟祯
李万根
张霄旦
ZHONG Zhen;LI Wangen;ZHANG Xiaodan(Department of Endocrinology,the Second Affiliated Hospital of Guangzhou Medical University,Guangzhou 510260,China)
出处
《实用医学杂志》
CAS
北大核心
2018年第24期4022-4026,共5页
The Journal of Practical Medicine
基金
广东省自然科学基金项目(编号:2017A030310257)
广州市科技计划项目(编号:201707010045)
广州医科大学博士
留学回国人员科研项目(编号:2015C10)
关键词
糖尿病心肌病
肿瘤坏死因子受体相关蛋白1
线粒体
氧化应激
diabetic cardiomyopathy
tumor necrosis factor receptor-associated protein 1
mitochondria
oxidative stress
