南昌大学学报(医学版)2024,Vol.64Issue(1) :20-24.DOI:10.13764/j.cnki.ncdm.2024.01.004

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

赵嘉仪 袁明明 王利军 赖松青 邹华西 黄琼 刘季春 黄璜
南昌大学学报(医学版)2024,Vol.64Issue(1) :20-24.DOI:10.13764/j.cnki.ncdm.2024.01.004

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

赵嘉仪 1袁明明 2王利军 2赖松青 2邹华西 2黄琼 2刘季春 2黄璜1
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作者信息

  • 1. 南昌大学,第一附属医院心脏大血管外科,南昌 330006;南昌大学,焕奎书院,南昌 330006
  • 2. 南昌大学,第一附属医院心脏大血管外科,南昌 330006
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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.

Abstract

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 H9 C2 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

Key words

voltage-dependent anionic channel protein 1(VDAC1)/myocardial cells/plasmid construction/adenovirus vector

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基金项目

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)

出版年

2024
南昌大学学报(医学版)
南昌大学

南昌大学学报(医学版)

CSTPCD
影响因子:1.008
ISSN:2095-4727
参考文献量14
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