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大鼠运动性肌损伤特征microRNA表达及其膜损伤调控靶点的预测

徐玉明, 曹建民, 李俊平, 黄巧婷, 王平

杭州师范大学体育与健康学院，杭州 310036；北京体育大学运动人体科学学院，北京 100086

摘要

本文旨在筛选大鼠运动性肌损伤(exercise-induced muscle damage, EIMD)的特征microRNA (miRNA)，并预测分析特征miRNA调控膜损伤的靶点。将24只健康雄性Sprague-Dawley (SD)大鼠平均分为安静对照(C)组、运动后24 h (E24)组、运动后48 h (E48)组，采取一次间歇性离心运动(−16°下坡跑台跑)复制EIMD模型，用伊文氏蓝荧光染色法鉴定EIMD特征，用基因芯片进行差异性miRNA筛选，用RT-qPCR验证芯片检测结果后获得EIMD差异性miRNA表达谱，RT-qPCR检测EIMD膜损伤相关特征蛋白和相关通路核心分子的mRNA表达水平并预测特征miRNA的靶基因。本研究筛选出并验证了2条EIMD特征miRNA：miR-206-3p和miR-139-3p。骨架蛋白dystrophin (r = −0.68)、utrophin (r = −0.64)和MAPK信号通路核心分子JNK (r = −0.62)、ERK1 (r = −0.68)均与miR206-3p呈中度负相关(P < 0.001)，而均与miR-139-3p无显著相关。以上研究结果提示，EIMD可诱导成年大鼠腓肠肌miRNA表达谱明显改变，miR-206-3p和miR-139-3p是EIMD特征miRNA，miR-206-3p可能通过调控骨架蛋白dystrophin、utrophin和MAPK信号通路分子JNK、ERK1调控膜损伤。

关键词： 运动; 骨骼肌 ; 损伤 ; 膜 ; 微小RNA

分类号：G804.2;R363

Analyses of exercise-induced muscle damage-specific microRNA expression and molecular target of sarcolemmal damage in rats

XU Yu-Ming, CAO Jian-Min, LI Jun-Ping, HUANG Qiao-Ting, WANG Ping

Sports and Health College of Hangzhou Normal University, Hangzhou 310036, China； Sport Science College of Beijing Sport University, Beijing 100086, China

Abstract

In the present study, we were to screen the specific microRNA (miRNA) of exercise-induced muscle damage (EIMD) and assess the EIMD-specific miRNAs-regulated target of sarcolemmal damage in rats. Twenty-four male Sprague-Dawley (SD) rats were randomly divided into 3 groups, which included sedentary (C), 24 h post-exercise (E24) and 48 h post-exercise (E48) groups. Rat EIMD model was established by an acute eccentric exercise, i.e., a downhill running treatment at −16&ordm; gradient. EIMD characteristics were verified by Evans blue dye staining, differentially expressed miRNAs were detected by microarray assay, EIMD-specific miRNAs expressions were further validated by real-time quantitative RT-PCR (RT-qPCR), and targets of the miRNAs were predicted based on mRNA expressions of associated proteins and related pathway core molecules of sarcolemmal damage. Two EIMD-specific expressed miRNAs, including miR-206-3p and miR-139-3p, were found in the study. There was a significantly negative correlation (P < 0.05) between miR-206-3p expression and dystrophin (r = −0.68), utrophin (r = −0.64), JNK (r = −0.62) or ERK1 (r = −0.68) respectively, but no correlation was found between miR-139-3p and these biomolecules. The results suggest that: i) the expression profile of miRNAs in rat is significantly affected by EIMD, ii) miR-206-3p and miR-139-3p are the EIMD-specific miRNAs, and iii) miR-206-3p may control sarcolemmal damage by regulating dystrophin, utrophin, JNK and ERK1.

Key words: Exercise; skeletal muscle ; damage ; sarcolemma ; microRNA

收稿日期：2016-12-01　　录用日期：2016-12-27

通讯作者：徐玉明　　E-mail: xuyuming110@126.com

引用本文：

徐玉明, 曹建民, 李俊平, 黄巧婷, 王平. 大鼠运动性肌损伤特征microRNA表达及其膜损伤调控靶点的预测[J]. 生理学报 2017; 69 (3): 276-284.

XU Yu-Ming, CAO Jian-Min, LI Jun-Ping, HUANG Qiao-Ting, WANG Ping. Analyses of exercise-induced muscle damage-specific microRNA expression and molecular target of sarcolemmal damage in rats. Acta Physiol Sin 2017; 69 (3): 276-284 (in Chinese with English abstract).