CAJ | 학술논문

目的筛选、分析与非瓣膜性心房颤动(房颤)相关的微RNA(miRNA).方法选择2009年1月至2010年12月北京安贞医院心外科的非瓣膜性房颤患者33例,应用低密度芯片技术,检测非瓣膜性房颤患者和健康人左心耳组织中miRNA表达谱的差异,用生物信息学方法筛选与房颤相关的miRNA及其靶基因,进一步通过定量反转录(RT)-PCR,Western印迹法和体外构建报告载体实验,验证miRNA和其靶基因之间的关系.结果与健康人比较,房颤患者左心耳组织中发现10个差异表达的miRNA:miR-155、miR-142-3p、miR-19b、miR-223、miR-146b-5p、miR-486-5p、miR-301b、miR-193b和miR-519b表达高,miR-193a-5p表达低(均p＜0.05)；其中miR-155表达差异最大(9.42±4.74比1.63±0.65),为健康人组织的5.78倍.miR-155的高表达导致了预测的靶基因CACNA1C和相应Cav1.2蛋白的低表达,体外构建报告载体实验发现miR-155可以结合到CACNA1C的3’-UTR区.结论非瓣膜性房颤患者左心耳组织中存在与其发病相关的miRNA,其中miR-155有可能通过调控Cav1.2的表达,参与了房颤的电重构.
목적 사선、분석여비판막성심방전동(방전)상관적미RNA(miRNA).방법 선택2009년1월지2010년12월북경안정의원심외과적비판막성방전환자33례,응용저밀도심편기술,검측비판막성방전환자화건강인좌심이조직중miRNA표체보적차이,용생물신식학방법사선여방전상관적miRNA급기파기인,진일보통과정량반전록(RT)-PCR,Western인적법화체외구건보고재체실험,험증miRNA화기파기인지간적관계.결과 여건강인비교,방전환자좌심이조직중발현10개차이표체적miRNA:miR-155、miR-142-3p、miR-19b、miR-223、miR-146b-5p、miR-486-5p、miR-301b、miR-193b화miR-519b표체고,miR-193a-5p표체저(균p＜0.05)；기중miR-155표체차이최대(9.42±4.74비1.63±0.65),위건강인조직적5.78배.miR-155적고표체도치료예측적파기인CACNA1C화상응Cav1.2단백적저표체,체외구건보고재체실험발현miR-155가이결합도CACNA1C적3’-UTR구.결론비판막성방전환자좌심이조직중존재여기발병상관적miRNA,기중miR-155유가능통과조공Cav1.2적표체,삼여료방전적전중구.
Objective To detect the differential expressions of miRNAs in left atrial appendage (LAA) in patients with atrial fibrillation (AF).Methods Left atrial samples were collected from nonvalvular AF patients and healthy controls.The miRNA transcriptome was analyzed by microarray and verified by real-time reverse transcription-polymerase chain reaction.Computational prediction identified the AF-related miRNAs and its target gene.In the meantime,construction of reporter plasmids and reporter assays were performed to test whether miRNA could represses the Luciferase activity of 3' untranslated regions of its target gene.Results MiR-155,miR-142-3p,miR-19b,miR-223,miR-146b-5p,miR-486-5p,miR-301b,miR-193b,miR-519b were found to be up-regulated by ＞2 folds whereas miR-193a-5p wasdown-regulated.In particular,the level of miR-155 increased by 5.78 folds in AF patients versus healthycontrols (9.42 ± 4.74 vs 1.63 ±0.65).Furthermore,computational prediction identified CACNAICencoding Cav1.2 as a direct target of miR-155.In the meantime,the construction of reporter plasmids andreporter assays showed that miR-155 repressed the Luciferase activity of 3' untranslated regions ofCACNAIC.Conclusion In LAA sample of nonvalvular AF,there is an expression of AF-related miRNAsincluding miR-155.And it reveals a potential link between the reglation of Cav1.2 and miR-155 in electricremodeling of AF.