主动脉弓缩窄模型所致小鼠主动脉重塑的lncRNA表达谱分析  

Expression profile of lncRNAs of aortic remodeling in a murine aortic arch constriction model

作　　者：冯婷 周风云 陈扬平 路祥麒 张秋霞[1] 陈业佳 张新禄[1] 修建成[1] Feng Ting;Zhou Feng-Yun;Chen Yang-Ping;Lu Xiang-Qi;Zhang Qiu-Xia;Chen Ye-Jia;Zhang Xin-Lu;Xiu Jian-Cheng(Department of Cardiology,Nanfang Hospital,Southern Medical University,Guangzhou 510515,China)

机构地区：[1]南方医科大学南方医院心血管内科,广州510515

出　　处：《解放军医学杂志》2020年第9期913-921,共9页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金青年科学基金(81800371);国家重点专项项目子课题(2018YFC1312803)。

摘　　要：目的分析主动脉弓缩窄(TAC)模型所致高压负荷诱导的小鼠主动脉重塑的长链非编码RNA(lncRNA)表达谱并探讨其可能的作用机制。方法取60只8周龄C57BL/6J雄性小鼠,分别构建TAC模型及进行假手术处理,术后1周TAC模型小鼠右颈总动脉/左颈总动脉血流速度比值为5~10时纳入实验组,假手术处理小鼠该比值为<1者纳入对照组,各24只。分别采用HE、Weigert及天狼猩红染色观察血管中膜厚度、中膜层弹力纤维及胶原纤维沉积情况。应用高通量测序技术鉴定差异表达转录本,并通过生物信息学分析的方法解析动脉重塑后转录本的表达变化。结果术后1周,实验组的右颈总动脉/左颈总动脉血流速度比值高于对照组,差异有统计学意义(P<0.001)。术后2周,与对照组相比,实验组升主动脉内径增大,且发生明显重塑,包括中膜增厚、中膜弹力纤维变稀疏以及大量胶原纤维沉积。根据建立的标准[差异倍数≥2,P<0.05,错误发现率(FDR值)<0.05],实验组与对照组在转录水平上共有199个lncRNA发生显著变化。任意挑选6个lncRNA进行qRT-PCR验证,与测序结果一致。预测lncRNA顺式作用与反式作用的靶基因,GO功能富集及KEGG通路富集分析表明,细胞外基质、细胞基质黏附、核因子(NF)-κB通路及血管平滑肌细胞收缩相关靶基因富集明显。结论本研究提供了高压负荷所致主动脉重塑中lncRNA表达变化的概况,提示lncRNA参与调节动脉重塑的细胞外基质合成过程,可能有助于阐明高压负荷所致主动脉重塑的潜在分子机制。Objective To study the expression profile and possible mechanism of long non-coding RNA(lncRNA)of aortic remodeling induced by elevated blood pressure in a murine transverse aortic arch constriction(TAC)model.Methods Sixty 8-week-old male C57BL/6J mice were randomly and averagely divided to receive TAC surgery or sham operation.TAC model mice were included in the experimental group when the ratio of blood flow velocity of right common carotid artery/left common carotid artery(RC/LC)was 5-10 at 1 week after operation,and mice treated with sham operation were included in the control group when RC/LC was less than 1.Twenty four mice for each group met the criteria in the end.HE staining,Weigert staining and Sirius red staining were used to observe the media wall thickness,elastic fiber and collagen fiber deposition of the ascending aorta.Two weeks after the surgery,aortic tissues were harvested for high throughput sequencing to identify differentially expressed lncRNA.Bioinformatics analysis were then performed to analyze the changes of lncRNA expression.Results One week after modeling,the ultrasound results showed that the blood flow velocity increased significantly at the ascending aorta in experimental group,and the blood flow velocity ratio(RC/LC)was higher significantly in experimental group than in control group(P<0.001).Two weeks after modeling and compared to that in control group,the inner diameter of the ascending aorta enlarged in experimental group and the tunica media of ascending aorta developed obvious remodeling,including the thickening of the tunica media,the thinning of media elastic fibers and the deposition of a large amount of collagen fibers.Using the well established cutoffs(fold change≥2,P<0.05 and false discovery rate(FDR)<0.05),a total of 199 lncRNAs were significantly changed in experimental group and control group at transcription level.Six differentially expressed lncRNAs were randomly selected for verifying the accuracy of sequencing data byusing qRT-PCR, which was consistent with the

关 键 词：高血压 动脉重塑 长链非编码RNA 高通量测序技术 生物信息学分析 

分 类 号：R544.1[医药卫生—心血管疾病]