沉默LncRNA H19通过调节神经生长因子抑制心肌梗死后交感神经重构  

Knockdowa of LncRNA H19 inhibits sympathetic nerve remodeling after myocardial infarction by regulating nerve growth factor

作　　者：张学丽 郑璐 王瑜[1] 王康 闫素华[1] ZHANG Xueli;ZHENG Lu;WANG Yu;WANG Kang;YAN Suhua(Department of Cardiology,Shandong Provincial Qianfoshan Hospital,Shandong University,Jinan 250014,Shandong,China)

机构地区：[1]山东大学附属千佛山医院心内科,山东济南250014

出　　处：《山东大学学报（医学版）》2021年第5期73-81,共9页Journal of Shandong University:Health Sciences

基　　金：国家自然科学基金(81870253);泰山学者岗位建设基金(闫素华);山东第一医科大学学术提升计划(2019QL012)。

摘　　要：目的探讨H19与神经生长因子(NGF)在心肌梗死(MI)后室性快速性心律失常(VAs)发生过程中的作用与调控关系。方法采用左冠状动脉结扎法建立大鼠MI模型。(1)将14只健康SD大鼠分为MI组和假手术组(sham组),每组7只,采用qRT-PCR法检测MI后7d长链非编码RNA H19(LncRNA H19)的表达;(2)将32只大鼠随机分为假手术+对照病毒组(sham+NC组)、假手术+沉默病毒组(sham+siH19组)、MI+对照病毒组(MI+NC组)、MI+沉默病毒组(MI+siH19组),每组8只,在MI手术当天,将H19沉默慢病毒或对照病毒注射到左心室心肌,在MI后7 d处死之前,进行电生理检查,取出心脏,分别采用Western blotting、qRT-PCR和免疫荧光等方法检测H19的表达及其对NGF、神经重构指标酪氨酸羟化酶(TH)、生长相关蛋白43(GAP43)及心律失常易感性的影响;(3)将24只大鼠随机分为MI+NC组、MI+siH19组和MI+siH19+LV-NGF组,每组8只,进行回复实验,采用Western blotting、免疫荧光法检测NGF、TH和GAP43的表达。结果大鼠MI 7 d后,H19表达上调(P<0.01);沉默H19后,免疫荧光结果显示,大鼠MI后TH、GAP43的阳染面积下降(P<0.001);程序性电刺激显示,MI后VAs的易感性降低(P<0.001);沉默H19可以有效抑制NGF的表达(Z=-2.402 P=0.016),而且沉默H19对交感神经重构的抑制作用也可被过表达的NGF改善(PTH<0.001,PGAP43=0.001)。结论MI后沉默H19可通过调节NGF的表达而抑制交感神经重构,从而降低MI后心律失常的发生率。Objective To explore the roles of H19 and nerve growth factor(NGF)in the development of ventricular arrhythmias(VAs)after myocardial infarction(MI).Methods MI rat models were established by left coronary artery occlusion.(1)A total of 14 healthy Sprague Dawley(SD)rats were randomly divided into the MI group and sham group,with 7 rats in each group.Seven days after MI was induced,the rats were sacrificed to determine the H19 expression with qRT-PCR.(2)A total of 32 rats were randomly divided into 4 groups:sham+NC group,sham+siH19 group,MI+NC group and MI+siH19 group,with 8 rats in each group.H19-siRNA or the control virus was injected into the left ventricular myocardium of the rats.Programmed electrical stimulation was performed 7 days after MI was induced to observe the susceptibility to VAs.The effects of H19 on NGF,tyrosine hydroxylase(TH)and growth-associated protein 43(GAP43)were detected with qRT-PCR,Western blotting and immunofluorescence staining.(3)A total of 24 rats were randomly divided into the MI+NC group,MI+siH19 group and MI+siH19+LV-NGF group for rescue test,with 8 rats in each group.The expressions of NGF,TH and GAP43 were detected with Western blotting and immunofluorescence staining.Results The expression of H19 were up-regulated 7 days after MI was induced(P<0.001).H19 knockdown suppressed the positive staining area of TH and GAP43(P<0.001),decreased the susceptibility to VAs(P<0.01),and inhibited the expression of NGF(Z=-2.402,P=0.016).The inhibitory effect of H19 knockdown on sympathetic remodeling could be saved by overexpressed NGF(PTH<0.001,PGAP43=0.001).Conclusion H19 knockdown after myocardial infarction can inhibit sympathetic nerve remodeling by regulating the expressiou of NGF,thus reducing the incidence of arrhythmia.

关 键 词：心肌梗死 交感神经重构 室性心律失常 LncRNA H19 神经生长因子 

分 类 号：R541[医药卫生—心血管疾病]