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目的探讨miR-1/133是否参与病毒性心肌炎心肌细胞钾离子、钙离子通道基因表达的调节。方法建立Balb/c小鼠急性病毒性心肌炎模型。将小鼠分为对照组、心肌炎组、心肌炎+miR-1/133mimics组、心肌炎+miR-1/133 NC组,每组10只。采用苏木精-伊红染色法观察心肌形态学改变;采用qRT-PCR法检测心肌中miR-1、miR-133及Kcnd2、Irx5、Kcnj2和α1c的相对表达量;采用Western blotting法检测心肌中蛋白Kv4.2、Kir2.1、Cav1.2的相对表达量。结果苏木精-伊红染色显示对照组心肌细胞排列整齐,间质无炎性细胞浸润;心肌炎组与心肌炎+miR-1/133 NC组心肌细胞水肿、排列紊乱,炎性细胞浸润间质;心肌炎+miR-1/133mimics组心肌细胞排列较整齐,无细胞水肿,间质少量炎性细胞浸润。与对照组相比,心肌炎组与心肌炎+miR-1/133 NC组心肌miR-1、miR-133及Kcnd2、Kcnj2表达下调,蛋白Kv4.2、Kir2.1的表达下调(P<0.01);Irx5、α1c及蛋白Cav1.2表达均上调(P<0.01);心肌炎+miR-1/133 mimics组较心肌炎组与心肌炎+miR-1/133 NC组相比,miR-1、miR-133及Kcnd2、Kcnj2表达上调,蛋白Kv4.2、Kir2.1表达上调(P<0.05),Irx5、α1c及蛋白Cav1.2表达均下调(P<0.01)。结论 miR-1/133参与病毒性心肌炎心肌细胞钾离子和钙离子通道基因表达的调节。
Objective To investigate whether miR-1/133 is involved in the regulation of K + and Ca 2+ channel gene expression in viral myocarditis. Methods Acute viral myocarditis in Balb / c mice was established. Mice were divided into control group, myocarditis group, myocarditis + miR-1 / 133mimics group, myocarditis + miR-1/133 NC group, 10 rats in each group. The myocardial morphological changes were observed by hematoxylin-eosin staining. The relative expression of miR-1, miR-133, Kcnd2, Irx5, Kcnj2 and α1c in myocardium was detected by qRT-PCR. The relative expression of protein Kv4.2, Kir2.1, Cav1.2. Results The hematoxylin-eosin staining showed that the myocardial cells in the control group were arranged neatly with no interstitial inflammatory cell infiltration. The myocardium edema, disordered structure and infiltration of inflammatory cells in myocarditis and myocarditis + miR-1 / Myocarditis + miR-1 / 133mimics group of myocardial cells arranged more neat, no cell edema, interstitial infiltration of a small amount of inflammatory cells. Compared with the control group, the expression of miR-1, miR-133, Kcnd2 and Kcnj2 in myocarditis group and myocarditis + miR-1 / 133NC group were down-regulated and the expressions of Kv4.2 and Kir2.1 proteins were down-regulated in myocarditis group and myocarditis group (P <0.01). The expression of miR-1, miR-133, and Cav1.2 in myocarditis + miR-1/133 mimics group were higher than those in myocarditis + miR-1/133 NC group The expressions of Kcnd2 and Kcnj2 were up-regulated, the expressions of Kv4.2 and Kir2.1 were upregulated (P <0.05), while the expressions of Irx5, α1c and Cav1.2 were down-regulated (P <0.01). Conclusion miR-1/133 is involved in the regulation of gene expression of potassium and calcium channels in viral myocarditis.