Uqcrc1的RNA干扰片段筛选及其对H9C2心肌细胞耐受缺氧/复氧损伤的影响

被引：3

作者：

吴潇潇

李洪

易婷婷

杨天德

机构：

[1] 第三军医大学附属新桥医院麻醉科

来源：

中华临床医师杂志(电子版) | 2013年 / 7卷 / 24期

关键词：

RNA干扰; Uqcrc1; H9C2心肌细胞; 缺氧/复氧;

D O I：

暂无

中图分类号：

R363 [病理生理学];

学科分类号：

100104 ;

摘要：

目的筛选泛醇-细胞色素c还原酶核心蛋白(Ubiquinol-cytochrome c reductase core protein 1,Uqcrc1)高效的RNA干扰片段,探讨Uqcrc1基因沉默对大鼠H9C2心肌细胞耐受缺氧/复氧(hypoxia/reoxygenation,H/R)损伤的影响。方法制备三种针对Uqcrc1的RNA干扰片段,采用RT-PCR和Western blot检测Uqcrc1 RNA干扰片段转染后Uqcrc1基因和蛋白的表达,从中筛选出最有效的RNA干扰片段及转染浓度,采用MTT法和LDH试剂盒分别检测H/R损伤后H9C2心肌细胞的存活率和LDH漏出率。结果靶序列为CCGUUGCUGUAGCUAACAAdTdT的siUqcrc1片段对Uqcrc1 mRNA的抑制作用最明显,在转染浓度为200 nmol/L时对Uqcrc1蛋白表达抑制最明显。靶向Uqcrc1的RNA干扰片段转染降低了H9C2心肌细胞耐受H/R损伤的能力。与阴性对照组相比,Uqcrc1基因沉默组H9C2心肌细胞H/R损伤后的存活率显著降低,LDH漏出率升高(P<0.01)。结论 Uqcrc1在心肌细胞耐受H/R损伤中起了重要作用,它可能是心肌保护的又一个关键靶点。

引用

页码：11469 / 11472

页数：4

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