容积激活性氯离子通道蛋白ClC-3表达下调促进H9c2心肌细胞肥大及其机制研究  

The volume-activated chloride channel protein ClC-3 down-expression promotes the hypertrophy of H9c2 cardiomyocytesand its mechanism

作　　者：孙宇 陈梦青 汪帅 李艳文 喻阳 李咏琪 李春梅 Sun Yu;Chen Mengqing;Wang Shuai(School of Life Sciences and Biopharmaceuticals, Guangdong Pharmaceutical University, Guangzhou 510006;Shenzhen Inustitute of Advanced Technology Chinese Academy of Science, Shenzhen 440305)

机构地区：[1]广东药科大学生命科学与生物制药学院,广州510006 [2]中国科学院深圳先进技术研究院,深圳440305

出　　处：《安徽医科大学学报》2021年第7期1075-1081,共7页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:31500926)。

摘　　要：目的探讨容积激活性氯离子通道蛋白ClC-3表达下调对H9c2心肌细胞的影响及其机制。方法用siRNA转染H9c2心肌细胞构建ClC-3表达下调的细胞模型;免疫荧光染色观察H9c2细胞表面积;qRT-PCR检测H9c2细胞心房利钠肽(ANP)、脑钠肽(BNP)和β-肌球蛋白重链(β-MHC)的mRNA表达水平;JC-1荧光探针检测线粒体膜电位(MMP)水平,图像分析系统研究H9c2心肌细胞的调节性体积回缩能力(RVD);全细胞膜片钳技术检测H9c2细胞膜上氯电流的变化。结果与空白组相比,siRNA转染H9c2细胞72 h能降低ClC-3 mRNA和蛋白的表达水平;而ClC-3下调与异丙肾上腺素相似,都增加H9c2心肌细胞表面积和心肌肥大标志性因子mRNA的表达,同时降低H9c2细胞的MMP和容积调节能力,抑制容积激活性氯离子电流的激活。结论ClC-3表达的下调能诱导H9c2心肌细胞肥大,其机制可能与其损伤线粒体功能、降低细胞容积调节能力和抑制容积激活性氯离子电流的激活有关。Objective To investigate the effect and mechanism of down-regulation of volume-activated chloride channel protein ClC-3 on H9c2 cardiomyocytes.Methods H9c2 cardiomyocytes were transfected with siRNA to construct a cell model of down-regulated ClC-3;immunofluorescence staining was used to observe the surface area of H9c2 cells;qRT-PCR was used to detect ANP,BNP andβ-MHC of H9c2 cells;the level of mitochondrial membrane potential was detected by JC-1 fluorescent probe;the image analysis system was used to study the regulatory volume decrease(RVD)of H9c2 cardiomyocytes;the change of chloride current on H9c2 cell membrane was detected by whole cell patch clamp technology.Results Compared with the control group,siRNA transfection of H9c2 cells for 72 h could reduce the expression levels of ClC-3 mRNA and protein;and the down-regulation of ClC-3 expression was similar to isoproterenol,which could increase the surface area of H9c2 cardiomyocytes and the iconic factors.Besides,it decreased the mitochondrial membrane potential of H9c2 cells,reduced the capacity of volume regulation,and inhibited the activation of volume-activated chloride ion current.Conclusion The down-regulation of ClC-3 expression induces H9c2 cardiomyocyte hypertrophy.Its mechanism may be related to its damage to mitochondrial function,reduction of cell volume regulation and inhibition of volume-activated chloride current activation.

关 键 词：CLC-3 H9C2细胞 心肌细胞肥大 线粒体膜电位 容积激活性氯电流 

分 类 号：R329.21[医药卫生—人体解剖和组织胚胎学]