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.
作者
赵嘉仪
袁明明
王利军
赖松青
邹华西
黄琼
刘季春
黄璜
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)
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)。
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
voltage-dependent anionic channel protein 1(VDAC1)
myocardial cells
plasmid construction
adenovirus vector
