大鼠VDAC1腺病毒过表达载体构建及功能鉴定  

Construction and Functional Characterization of Rat VDAC1 Adenovirus Overexpression Vector

作　　者：赵嘉仪 袁明明 王利军 赖松青[1] 邹华西 黄琼[1] 刘季春[1] 黄璜[1,2] ZHAO Jia-yi;YUAN Ming-ming;WANG Li-jun;LAI Song-qing;ZOU Hua-xi;HUANG Qiong;LIU Ji-chun;HUANG Huang(Department of Cardiac and Macrovascular Diseases,the First Affiliated Hospital,Nanchang University,Nanchang 330006,China;Huankui Academy,Nanchang University,Nanchang 330006,China)

机构地区：[1]南昌大学第一附属医院心脏大血管外科,南昌330006 [2]南昌大学焕奎书院,南昌330006

出　　处：《南昌大学学报（医学版）》2024年第1期20-24,共5页Journal of Nanchang University：Medical Sciences

基　　金：江西省中医药管理局科技计划(2022A355);南昌大学“四新”研究与改革实践项目(NCUJGLX-2022-160-130);南昌大学创新创业教育类教学改革研究项目(NCUSCJG-2022N58)。

摘　　要：目的构建电压依赖性阴离子通道蛋白1(VDAC1)腺病毒过表达载体,并观察其在H9C2细胞中的表达情况,为探究VDAC1在心肌细胞缺血再灌注损伤中的作用机制奠定基础。方法构建ADV6-NC腺病毒及ADV6-VDAC1腺病毒,取第6代H9c2心肌样细胞用于转染。转染细胞分为3组:空白对照组、阴性对照组(转染ADV6-NC腺病毒)和实验组(转染ADV6-VDAC1腺病毒)。通过qPCR和蛋白免疫印迹实验检测各组细胞VDAC1的表达情况。结果成功构建ADV6-NC及ADV6-VDAC1,实验组的VDAC1表达量明显高于空白对照组和阴性对照组,差异具有统计学意义(P<0.05)。结论利用腺病毒载体可使H9c2细胞在体外持续高效表达VDAC1。Objective In order to lay a foundation for exploring the mechanism of action of voltage-dependent anionic channel protein 1(VDAC1)in myocardial ischemia-reperfusion injury,an adenovirus vector overexpressing VDAC1 was constructed and its expression in H9C2 cells was observed.Methods ADV6-NC adenovirus and ADV6-VDAC1 adenovirus were constructed,and H9c2 myocardial cells of the 6th generation were used for transfection.The cells were divided into three groups:blank control group,negative control group(transfection with ADV6-NC)and experimental group(transfection with ADV6-VDAC1).The expression of VDAC1 was detected by qPCR and Western blot.Results The ADV6-NC and ADV6-VDAC1 vectors were constructed successfully.The expression of VDAC1 in the experimental group was obviously higher than that in the blank control group and the negative control group(P<0.05).Conclusion H9c2 cells can express VDAC1 constantly and efficiently by using adenovirus vector in vitro.

关 键 词：电压依赖性阴离子通道蛋白1 肌细胞 质粒构建 腺病毒载体 

分 类 号：R332[医药卫生—人体生理学] R602[医药卫生—基础医学]