MicroRNA-328 Contributes to Adverse Electrical Remodeling in Atrial Fibrillation
MicroRNA-328 Contributes to Adverse Electrical Remodeling in Atrial Fibrillation
复制标题
DOI：
10.1161/circulationaha.110.958967
复制
发表时间：
2010-12-07
期刊：
影响因子：
37.8
通讯作者：
Yang, Baofeng
中科院分区：
文献类型：
--
作者：
Lu, Yanjie;Zhang, Ying;Yang, Baofeng
背景 - 临床和实验性心房颤动（AF）的一个特征是心房电重构，其与L型钙电流大幅降低以及动作电位时程缩短相关。微小RNA（miRNA）可能参与这一过程的可能性尚未得到验证。因此，我们评估了miRNA在调节实验性AF中的潜在作用。
方法和结果 - 通过微阵列分析miRNA转录组，并通过实时逆转录聚合酶链反应对来自右心房快速起搏8周建立AF的犬的左心房样本以及来自风湿性心脏病AF患者的人心房样本进行验证。发现miR - 223、miR - 328和miR - 664上调超过2倍，而miR - 101、miR - 320和miR - 499至少下调50%。特别是，相对于非AF受试者，miR - 328水平在AF犬中升高3.9倍，在AF患者中升高3.5倍。计算机预测确定CACNA1C和CACNB1（分别编码心脏L型钙通道α1c和β1亚基）为miR - 328的潜在靶点。通过腺病毒感染在犬心房中强制表达miR - 328以及在小鼠中采用转基因方法重现了AF的表型，例如AF易感性增强、L型钙电流降低以及心房动作电位时程缩短。用抗miR使miR - 328水平正常化可逆转这些情况，并且内源性miR - 328的基因敲低减弱了AF易感性。通过蛋白质印迹和荧光素酶活性测定证实CACNA1C和CACNB1为miR - 328的同源靶基因，显示出miR - 328水平与L型钙通道蛋白亚基之间的相互关系。
结论 - miR - 328通过靶向L型钙通道基因促进AF中的不良心房电重构。因此，该研究揭示了AF的一种新的分子机制，并表明miR - 328是AF的一个潜在治疗靶点。（《循环》2010年；122:2378 - 2387）
Background-A characteristic of both clinical and experimental atrial fibrillation (AF) is atrial electric remodeling associated with profound reduction of L-type Ca2+ current and shortening of the action potential duration. The possibility that microRNAs (miRNAs) may be involved in this process has not been tested. Accordingly, we assessed the potential role of miRNAs in regulating experimental AF.Methods and Results-The miRNA transcriptome was analyzed by microarray and verified by real-time reverse-transcription polymerase chain reaction with left atrial samples from dogs with AF established by right atrial tachypacing for 8 weeks and from human atrial samples from AF patients with rheumatic heart disease. miR-223, miR-328, and miR-664 were found to be upregulated by >2 fold, whereas miR-101, miR-320, and miR-499 were downregulated by at least 50%. In particular, miR-328 level was elevated by 3.9-fold in AF dogs and 3.5-fold in AF patients relative to non-AF subjects. Computational prediction identified CACNA1C and CACNB1, which encode cardiac L-type Ca2+ channel alpha 1c- and beta 1 subunits, respectively, as potential targets for miR-328. Forced expression of miR-328 through adenovirus infection in canine atrium and transgenic approach in mice recapitulated the phenotypes of AF, exemplified by enhanced AF vulnerability, diminished L-type Ca2+ current, and shortened atrial action potential duration. Normalization of miR-328 level with antagomiR reversed the conditions, and genetic knockdown of endogenous miR-328 dampened AF vulnerability. CACNA1C and CACNB1 as the cognate target genes for miR-328 were confirmed by Western blot and luciferase activity assay showing the reciprocal relationship between the levels of miR-328 and L-type Ca2+ channel protein subunits.Conclusions-miR-328 contributes to the adverse atrial electric remodeling in AF through targeting L-type Ca2+ channel genes. The study therefore uncovered a novel molecular mechanism for AF and indicated miR-328 as a potential therapeutic target for AF. (Circulation. 2010;122:2378-2387.)