证实microRNA-214参与慢性心衰大鼠心肌细胞钙调控过程,并揭示其内在作用机制.冠脉结扎法诱导大鼠慢性心衰模型.实时定量PCR结果显示与假手术组比,心衰组大鼠左心室miR-214表达显著上调(P〈0.05),Western blot实验法结果显示与假手术组比,心衰组大鼠左心室L-型钙通道β1亚基(Cavβ1)表达显著下调(P〈0.05).左心室局部肌肉注射agomiR-214导致假手术组和心衰组大鼠左心室收缩压(LVSP)、L-型钙通道电流密度、L-型钙通道β1亚基(Cavβ1)表达均显著降低(P〈0.05).荧光素酶报告基因实验结果显示miR-214能够调节心肌细胞L-型钙通道β1亚基基因CACNB1的表达.结果提示microRNA-214参与慢性心衰大鼠心肌细胞钙调控过程,其机制可能与microRNA-214抑制心肌细胞L-型钙通道β1亚基(Cavβ1)表达有关.
The goal of this study was to determine the role of microRNA-214 involoved in the calcium handling of cardiomyocytes in rats with chronic heart failure(CHF) and clarify the underlying mechanism. CHF animal model was induced by coronary artery ligation. The results of real-time PCR showed that microRNA-214 expression levels in the cardiomyocytes were significantly increased in CHF rats compared with sham rats. The results of Western blot revealed that L-type calcium channel subunit beta-1 (Cavl31) protein expression levels in the cardiomyocytes were significantly decreased in CHF rats compared with sham rats. Local intramuscular injections of agomiR-214 produced a significantly decrease in left ventricular systolic pressure (LVSP), the density of L- type calcium current and Cavl31 protein expression levels in both sham and CHF rats. Luciferase Reporter Assay demonstrated that miR-214 can down-regulate its target protein Cavl31. Our study suggested that a mechanism by which microRNA-214 involved in the calcium handling in CHF may be mediated through downregulating Cavl31 protein expression levels.