房颤射频消融术逆转冠状窦血miRNA异常表达的研究  

The radiofrequency ablation my reverse the abnormal miRNA expression in coronary sinus blood of patients with atrial fibrillation

作　　者：徐桂玉[1] 赵楠楠[1] 杨水祥[1] XU Gui-yu ZHAO Nan-nan YANG Shui-xiang.(Department of Cardiology, the Capital Medical University Affiliated Beijing Shijitan Hospital, Beijing 100038, China)

机构地区：[1]首都医科大学附属北京世纪坛医院心内科,北京市100038

出　　处：《中国心血管病研究》2016年第8期720-726,765,共8页Chinese Journal of Cardiovascular Research

摘　　要：目的 探讨房颤射频消融手术（RFA）终止房颤后患者冠状窦血miRNA的变化,试图发现真正起调控作用的miRNA,以揭示房颤的机制及可能的干预靶点.方法 选择30例行房颤射频消融术患者（阵发性、持续性和永久性房颤各10例）,健康体检者10例作为正常对照组.射频消融术前分别取冠状窦血和外周血,术后3个月取外周血,使用miRNA芯片进行全基因组miRNA表达谱微阵列分析,Real-time PCR对miRNA结果进行验证,并通过mirbase、miranda、targetscan数据库行靶基因分析,对重要miRNA进行双荧光素酶结合实验.结果 房颤射频消融术前患者冠状窦血与自身外周血比较,共有142种miRNA表达差异,其中6种显著上调、8种下调（P＜0.05）.射频消融术后外周血较术前上调的6种miRNA中分别有3种表达上调和下调,其中miR-1266下调-204.17倍;较术前下调的8种miRNA中,有7种再下调,其中miR-3664-5p下调-44.66倍.荧光素酶结合实验证实SCN5A是miR-1266的直接靶基因,CACNA1C是miR-4279的直接靶基因.结论 房颤射频消融手术可逆转患者冠状窦血miRNA的调控异常,冠状窦血miRNA的表达差异可直接反映房颤时心肌miRNA的表达状况.MiR-1266有可能成为未来房颤干预的靶点.Objective To study the effect of radiofrequency ablation（RFA ） on miRNA differential expres- sion in coronary sinus blood（SB ） of atrial fibrillation（AF ） patients, and try to identify the possible pathogenesis of AF and miRNA targets for intervention in future. Methods 30 AF patients （10 paroxymal, lO persistent and 10 permanent AF patients ） were enrolled, alongside 10 healthy subjects as controls. Peripheral blood （PB） samples were obtained before and at 3 months after RFA respectively. The total RNA was extracted and hybridized with the microRNA chips （microRNA v 18.0）, and the differential expression of miRNA and clustering analysis in whole genome were made with Volcano Plot and tMEV software respectively, and validated by Real-time PCR. The target gene analysis of miRNAs was predicted through the Mirbase, Miranda and Targetscan databases. The important miRNAs were made for dual luciferase binding experiment to analyze and validate target genes. Results There were 142 miRNAs differential expression in SB of pre-operation AF patients compared with PB of patient-self control, in which, 6 miRNAs were increased and 8 miRNAs were decreased significantly （P〈0.05）. Then, in the compare of PB in AF patients after RFA with which of self-control before RFA, there were 3 miRNAs up-regulat- ed and down-regulated respectively in above increased 6 miRNAs, while miR-1266 was down-regulated 204.17- fold. In above decreased 8 miRNAs, 7 of them were down-regulated again after RFA, while miR-574-3p up-regulated 5.25-fold. SCN5A was the target gene of miR-1266, and CACNAlC was the target gene of miR-4279 confirmed with Luciferase binding experiments. Conclusion RFA can reverse the abnormal expressions of miRNAs in AF patients. The differential expression of miRNAs in SB can directly reflect the regulatory status of miRNAs in cardiomyocytes in AF attack. MiR-1266 may become the future target for AF intervention.

关 键 词：心房颤动 射频消融 微小RNA 离子通道蛋白 组学研究 

分 类 号：R541.7[医药卫生—心血管疾病]