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乳鼠心肌细胞上清液对大鼠MSCs心肌动作电位相关离子通道基因表达的影响
Effect of supernatant of neonatal rat cardiomyocytes on gene expression of action potential-related ion channels in rat MSCs
【摘要】 目的观察不同状态下培养的乳鼠心肌细胞上清液对大鼠骨髓间充质干细胞(MSCs)心肌动作电位相关离子通道基因表达的影响。方法分别取大鼠骨髓MSCs和乳鼠心肌细胞进行体外培养。实验分为MSCs组、5-氮胞苷(5-Aza)诱导组、正常诱导组、缺氧诱导组和心肌细胞组,MSCs组为正常培养的MSCs,5-Aza诱导组为心肌定向诱导剂5-Aza干预的MSCs,正常诱导组为正常心肌细胞上清液干预的MSCs,缺氧诱导组为缺氧状态下的心肌细胞上清液干预的MSCs,心肌细胞组为不予干预的正常心肌细胞。采用RT-PCR法检测各组细胞膜上钠通道(SCN2a)、钾通道(Kv2.1、Kv1.4、Kir1.1、EAG1)和钙通道基因(CCHL2α)mRNA表达的变化。结果 MSCs组细胞膜上表达与心肌动作电位相关的离子通道基因。与MSCs组相比,其余各组SCN2a、Kv2.1、Kir1.1、EAG1、CCHL2α mRNA表达均升高(P<0.05或<0.01),且缺氧诱导组、心肌细胞组Kv1.4 mRNA表达升高(P<0.05或<0.01),但5-Aza诱导组、正常诱导组、缺氧诱导组各种离子通道基因表达两两比较差异无统计学意义。结论乳鼠正常心肌细胞上清液和缺氧心肌细胞上清液可诱导大鼠MSCs心肌动作电位相关离子通道基因表达增加,心肌微环境有利于大鼠MSCs发生心肌电生理变化。
【Abstract】 Objective To observe the effects of neonatal rat cardiomyocyte supernatants at different state on the gene expression of action potential-related ion channels in rat bone marrow mesenchymal stem cells( MSCs). Methods Rat bone marrow MSCs and neonatal rat cardiomyocytes were cultured in vitro. The cells were divided into the MSCs group,5-azacytidine( 5-Aza) induction group,normal induction group,hypoxia-induced group,and cardiomyocyte group. The MSCs group was normally cultured MSCs. The MSCs in the 5-Aza group were induced by myocardial inducer 5-Aza,the MSCs in the normal induction group was induced by normal cardiomyocyte supernatant,the MSCs in the hypoxia-induced group were induced by hypoxic cardiomyocyte supernatant,and the MSCs in the cardiomyocyte group were normal myocardial cells without induction. The mRNA expression of sodium( SCN2a),potassium( Kv2. 1,Kv1. 4,Kir1. 1,EAG1),and calcium( CCHL2α) channels in the cell membranes of each group was detected by RT-PCR. Results The ion channel genes associated with cardiac action potentials were expressed on the membrane of MSCs. Compared with the MSCs group,the mRNA expression of SCN2a,Kv2. 1,Kir1. 1,EAG1,and CCHL2α increased in the other groups( P < 0. 05 or P <0. 01),and the expression of Kv1. 4 mRNA in the hypoxia-induced group and cardiomyocyte group increased( P < 0. 05 or P < 0. 01). However,there was no significant difference in the expression of various ion channel genes between the 5-Aza induction group,the normal induction group,and the hypoxia-induced group. Conclusion The expression of ion channelrelated genes increases in MSCs after induction of normal and hypoxia cardiomyocyte supernatants,which shows that the myocardial microenvironment contributes to the myocardial electrophysiological changes of MSCs.
【Key words】 bone marrow-derived mesenchymal stem cells; cardiomyocytes; ionic channel; action potential;
- 【文献出处】 山东医药 ,Shandong Medical Journal , 编辑部邮箱 ,2018年22期
- 【分类号】R54
- 【下载频次】100