miR-208b对小鼠HL-1房颤细胞钙超载的影响及其机制分析  

Effect and mechanism of miR-208b on calcium overload in mouse HL-1 atrial fibrillation cells

作　　者：张卫卫[1] 石昭昭[1] 蔡琴[1] 赵黎[1] 刘婷婷[1] 李晨辉 ZHANG Wei-wei;SHI Zhao-zhao;CAI Qin(Department of Cardiology,Xi'an First Hospital,Xi'an Shaanxi 710002,China)

机构地区：[1]西安市第一医院心内科

出　　处：《临床和实验医学杂志》2019年第20期2183-2186,共4页Journal of Clinical and Experimental Medicine

基　　金：陕西省社会发展科技攻关项目(编号:2015SF227)

摘　　要：目的探讨miR-208b对小鼠HL-1房颤细胞钙超载的影响,并分析其作用机制。方法将小鼠HL-1心房肌细胞随机分为3组:对照组、起搏组和miR-208b组。后两组建立快速起搏HL-1房颤细胞模型,然后将miR-208b模拟物转染至miR-208b组细胞,将模拟物阴性对照转染至起搏组细胞,对照组不处理。全细胞膜片钳技术检测动作电位时程(APD)及L型钙电流(I Ca,L)的电流密度,实时荧光定量PCR及Western Blot检测钙通道蛋白mRNA及蛋白的表达。结果与对照组相比,起搏组和miR-208b组细胞中miR-208b表达显著增加(P<0.05),且miR-208b组高于起搏组(P<0.05)。与对照组相比,起搏组和miR-208b组APD50、APD90、I Ca,L的电流密度、CACNA1C mRNA、CACNB2 mRNA、CaV1.2蛋白、SERCA2 mRNA及SERCA2蛋白的表达均显著降低(P<0.05),且miR-208b组低于起搏组(P<0.05)。结论miR-208b在小鼠HL-1房颤细胞中高表达,其可通过抑制L型钙通道蛋白的表达和功能以及抑制肌浆网Ca 2+摄入,导致胞内Ca 2+超载,诱发AF。Objective To investigate the effect of miR-208b on calcium overload in mouse HL-1 atrial fibrillation cells and analyze its mechanism of action.Methods Mouse HL-1 atrial myocytes were randomly divided into 3 groups:control group,pacing group and miR-208b group.The cells in latter two groups were established a rapid pacing HL-1 atrial fibrillation cell model,then miR-208b mimic was transfected into the miR-208b group,and mock negative control was transfected into the pacing group,and the cells in control group were given no transfection.The whole cell patch clamp technique was used to detect the current density of action potential duration(APD)and L-type calcium current(I Ca,L).The expressions of calcium channel proteins in mRNA and protein levels were detected by real-time fluorescent quantitative PCR and Western Blot.Results Compared with the control group,the expression of miR-208b was significantly increased in the pacing group and the miR-208b group(P<0.05),and the miR-208b group was higher than the pacing group(P<0.05).Compared with the control group,the APD50,APD90,current density of I Ca,L and expressions of CACNA1C mRNA,CACNB2 mRNA,CaV1.2 protein,SERCA2 mRNA and SERCA2 protein in the pacing group and miR-208b group were significantly reduced(P<0.05),and the miR-208b group was lower than the pacing group(P<0.05).Conclusion miR-208b is highly expressed in HL-1 atrial fibrillation cells,which induces AF by inhibiting the expression and function of L-type calcium channel protein and inhibiting sarcoplasmic reticulum Ca 2+uptake,resulting in intracellular Ca 2+overload.

关 键 词：小鼠 房颤细胞 miR-208b 钙超载 机制 

分 类 号：R54[医药卫生—心血管疾病]