LncRNA-CCRR通过调控UBA6影响钠通道泛素化进而调控心梗后心律失常  

LncRNA-CCRR regulates arrhythmia induced by myocardial infarction by affecting sodium channel ubiquitination via UBA6

作　　者：孙飞涵 李聃宁 杨华 王圣洁 罗惠珊 郭建军 宣立娜 孙丽华 SUN Fei-han;LI Dan-ning;YANG Hua;WANG Sheng-jie;LUO Hui-shan;GUO Jian-jun;XUAN Li-na;SUN Li-hua(Dept of Pharmacology,College of Pharmacy,Harbin Medical University,Harbin 150081,China)

机构地区：[1]哈尔滨医科大学药学院药理学教研室,黑龙江哈尔滨150081

出　　处：《中国药理学通报》2024年第8期1437-1446,F0001,共11页Chinese Pharmacological Bulletin

基　　金：黑龙江省自然科学基金资助项目(No.LH2022H002)。

摘　　要：目的探讨心梗后心律失常钠通道泛素化的调节机制,深入研究lncRNA-CCRR在心肌梗死后调控心脏电生理重构的机制,对心梗后心律失常的防治具有重要意义。方法应用lncRNA-CCRR转基因鼠,C57BL/6小鼠在体注射lncRNA-CCRR过表达腺相关病毒,感染4周后结扎冠状动脉左前降支12 h构建小鼠急性心肌梗死模型,程序性电刺激检测心律失常发生率。将lncRNA-CCRR过表达/敲减腺相关病毒、阴性对照转染至原代乳鼠心肌细胞中,缺氧12 h。FISH检测lncRNA-CCRR表达,Western blot和实时荧光定量PCR检测Na_(v)1.5和UBA6蛋白以及Na_(v)1.5的mRNA表达,免疫荧光检测Na_(v)1.5和UBA6表达,RIP检测lncRNA-CCRR与UBA6作用关系,全细胞膜片钳检测细胞CCRR过表达及敲减后I_(Na)电流密度。结果心梗小鼠心肌梗死边缘区lncRNA-CCRR表达降低,心律失常发生率升高,CCRR和Na_(v)1.5 mRNA表达下调,Na_(v)1.5蛋白表达下调,UBA6蛋白表达上调;过表达CCRR可逆转上述变化。转染AAV-CCRR可逆转缺氧后下调的CCRR和Na_(v)1.5 mRNA水平,改善Na_(v)1.5和UBA6蛋白表达,且lncRNA-CCRR与UBA6蛋白具有相互结合作用;转染AAV-CCRR后I_(Na)密度增加;转染AAV-si-CCRR后I_(Na)密度降低。结论心梗后lncRNA-CCRR表达降低;lncRNA-CCRR可通过抑制UBA6,增加Na_(v)1.5蛋白表达和I_(Na)电流密度改善心梗后心律失常。Aim To investigate the regulatory mechanism of arrhythmia of sodium channel ubiquitination after MI and to study the electrophysiological remodeling mechanism of lncRNA-CCRR after MI for the prevention and treatment of arrhythmia after MI.Methods LncRNA-CCRR transgenic mice and C57BL/6 mice injected with lncRNA-CCRR overexpressed adeno-associated virus were used.Four weeks after infection,the left anterior descending branch of the coronary artery was ligated for 12 h to establish a mouse acute myocardial infarction model,and the incidence of arrhythmia was detected by programmed electrical stimulation.LncRNA-CCRR overexpression/knockdown adeno-associated virus and negative control were transfected into neonatal mouse cardiomyocytes(NMCMs),and the model was prepared by hypoxia for 12 h.LncRNA-CCRR expression was detected by FISH,Na_(v)1.5 and UBA6 protein and Na_(v)1.5 mRNA expression were detected by Western blot and real-time quantitative polymerase chain reaction(qRT-PCR),Na_(v)1.5 and UBA6 expressions were detected by immunofluorescence,and the relationship between lncRNA-CCRR and UBA6 was detected by RIP.I_(Na)current density after CCRR overexpression and knockdown was detected by Whole-cell clamp patch.Results In MI mice,the expression of lncRNA-CCRR decreased,the incidence of arrhythmia increased,the expression of CCRR and Na_(v)1.5 mRNA was down-regulated,the protein expression of Na_(v)1.5 was down-regulated,and the protein expression of UBA6 was up-regulated compared with sham group.Overexpression of CCRR could reverse the above changes.AAV-CCRR could reverse the down-regulated CCRR and Na_(v)1.5 mRNA levels after hypoxia,and improve the expression of Na_(v)1.5 and UBA6 protein.The direct relationship between lncRNA-CCRR and UBA6 was identified by RIP analysis.The I_(Na)density increased after transfection with AAV-CCRR.The I_(Na)density decreased after transfection with AAV-si-CCRR.Conclusions The expression of lncRNA-CCRR decreases after MI,and lncRNA-CCRR can improve arrhythmia induced by MI by inhibi

关 键 词：心肌梗死 lncRNA-CCRR UBA6 钠通道 泛素化 心律失常 

分 类 号：R-332[医药卫生] R329.24R342.2R541.7R542.22