miRNAs可能通过调控MAPK通路参与心室间隔缺损  

miRNAs may involve in ventricular septal defect through regulating the MAPK pathway

作　　者：江辉 张林[1] JIANG Hui;ZHANG Lin(College of Life Sciences, Zhejiang Chinese Medicial University, Hangzhou 310053, China)

机构地区：[1]浙江中医药大学生命科学学院,浙江杭州310053

出　　处：《基础医学与临床》2022年第4期590-598,共9页Basic and Clinical Medicine

基　　金：浙江省教育厅高校国内访学者教师专业发展项目(FX2019021);浙江中医药大学科研创新人才项目(KC201921)。

摘　　要：目的探讨miRNAs在心室间隔缺损(VSD)中发挥的作用。方法通过miRNAs表达谱芯片实验分析心室间隔缺损(VSD)组和健康组全血样本中的差异表达miRNAs,通过生物信息数据库miRbase、Targetscan和Miranda进行靶基因测预。通过KEGG和REACTOME通路数据库筛选靶基因信号通路;采用RT-qPCR和Western blot检测关键信号通路中关键基因转录和蛋白表达水平。结果实验共得到22例差异表达miRNAs,预测获得12744个靶基因;共得到126条通路,对其中与心脏发育密切相关的MAPK通路进行了深入分析,其中ERK子通路中的RASA1、NF1的转录和蛋白表达水平均明显增加(P<0.05);Ras的转录和蛋白表达水平均明显降低(P<0.05);ERK、MEK1磷酸化蛋白表达水平显著下降(P<0.05);p38/MAPK子通路中的TNF、TNFRSF1A、TRAF2的转录和蛋白表达水平均明显增加(P<0.05);p38磷酸化蛋白表达水平显著上升(P<0.05);MEF2C的转录、总蛋白及磷酸化蛋白表达水平均明显上升(P<0.05)。上述通路受到12个miRNAs调控。结论22个差异miRNAs表达异常,导致多种已知心脏发育相关基因表达紊乱;其中12个表达异常miRNAs抑制ERK通路进而抑制心肌细胞分化、过度激活p38/MAPK通路表达进而促进心肌细胞异常凋亡,最终导致VSD的发生。Objective To investigate the role of miRNAs in pathogenesis of ventricular septal defect.MethodsThe miRNAs differential expression in whole blood samples from ventricular septal defect(VSD)group and normal group was analyzed by miRNAs expression microarray experiment and the target genes were predicted by biological information databases miRbase,Targetscan and Miranda.Target gene signaling pathways were screened through KEGG and REACTOME pathway databases,the transcription and protein expression of key genes in key signaling pathways were detected by RT-qPCR and Western blot.Results A total of 22 cases of differentially expressed miRNA were obtained in the experiment,and 12744 target genes were predicted.A total of 126 pathways were found and the MAPK pathway,which was closely related to heart development,and was deeply analyzed.Among them,the transcription and protein expression of RASA1 and NF1 in ERK pathway were significantly increased(P<0.05),the transcription and protein expression of RAS were significantly decreased(P<0.05),and the expression levels of ERK and MEK1 phosphorylated proteins were also significantly decreased(P<0.05);the transcription and protein expression of TNF,TNFRSF1A and TRAF2 in the p38/MAPK sub-pathway were significantly increased(P<0.05),the expression of p38 phosphorylated proteins was significantly increased(P<0.05),The transcrip-tion,total protein and phosphorylated protein expression of MEF2C were significantly increased(P<0.05).The above pathways were found to be regulated by 12 miRNA.Conclusions The abnormal expression of 22 differentially expressed miRNAs leads to a variety of known cardiac development-related gene expression disorders.Among them,12 abnormally expressed miRNAs inhibit the ERK pathway and thus inhibit the differentiation of cardiomyocytes and over-activate the expression of p38/MAPK pathway to promote the abnormal apoptosis of cardiomyocytes,and ultimately lead to the occurrence of VSD.

关 键 词：心室间隔缺损 差异表达miRNAs MAPK通路 

分 类 号：Q71[生物学—分子生物学]