沉默MagT1影响大鼠心肌细胞内Mg^(2+)水平和细胞凋亡的研究  

Effect of silencing MagT1 on the content of magnesium ion and apoptosis in rat cardiomyocytes

作　　者：滕飞翔 沈之君[2] 赵洪侠 张栋梁 杨留才 

机构地区：[1]江苏医药职业学院基础医学部,江苏盐城224005 [2]南通大学医学院病理学与病理生理学系,江苏南通226001 [3]潍坊护理职业学院医学基础部,山东潍坊261041

出　　处：《重庆医学》2018年第3期340-345,共6页Chongqing medicine

基　　金：江苏省盐城市医学科技发展计划项目(YK2016046)

摘　　要：目的使用siRNA沉默大鼠心肌细胞的镁离子转运蛋白1(MagT1),检测细胞内Mg^(2+)浓度变化和细胞凋亡情况。方法设计并合成MagT1siRNA序列,转入原代培养大鼠心肌细胞沉默MagT1,使用反转录PCR和Western blot检测MagT1mRNA和蛋白表达情况,荧光显微镜检测细胞内Mg^(2+)浓度变化情况,流式细胞法检测细胞凋亡情况。结果相比阴性siRNA组,MagT1siRNA转染大鼠心肌细胞48h,MagT1mRNA的沉默效率为51.83%(P<0.05),MagT1蛋白的沉默效率为56.75%(P<0.05),胞内Mg^(2+)浓度降低29.13%(P<0.05),细胞凋亡率为31.18%(P<0.01);转染60h,MagT1mRNA的沉默效率为86.91%(P<0.01),MagT1蛋白质的沉默效率为83.85%(P<0.01),细胞内Mg^(2+)浓度降低41.32%(P<0.01),细胞凋亡率为40.61%(P<0.01)。结论 MagT1被显著沉默后,细胞内Mg^(2+)浓度显著降低,细胞凋亡显著提高,细胞生命活动受到较大影响。Objective The changes of Mg^2＋ concentration and cell apoptosis were detected by using siRNA to silence MagTt in rat cardio myocytes. Methods MagT1 siRNA sequences were designed and synthetized,then transfected into primary cultured rat myocardial cell for silencing MagT1. The expression of MagT1 mRNA and protein were detected by RT-PCR and Western blot. The changes of Mg^2＋ concentration in the cells were detected by fluorescence microscopy. Flow cytometry （FCM） was used to detect the cell apoptosis. Results Compared with negative siRNA group,MagT1 siRNA transfected rat cardiomyocytes after 48 h, MagT1 mRNA silence efficiency was 51.83 % （P〈0.05）, the silence of MagT1 protein efficiency was 56.75 % （P〈0.05）, intracellular Mg^2＋ concentration was reduced by 29.13 % （P〈0.05） ,the apoptosis rate was 31.18 % （P〈0.01） ;MagT1 siRNA transfected rat cardiomyocytes after 60 h,MagT1 mRNA silence efficiency was 86.91% （P〈0.01）,the silence of MagT1 protein efficiency was 83.85% （P〈0. 01）, intracellular Mg^2＋ concentration was reduced by 41.32% （P〈0.01）, the apoptosis rate was 40.61% （P〈0.01）. Conclusion After the silencing of MagT1,the concentration of Mg^2＋ in the cells decreased significantly,the apoptotic rate increased significantly,cell life activities are greatly affected.

关 键 词：细胞凋亡 肌细胞 心脏 镁 镁离子转运蛋白1 SIRNA 

分 类 号：R331[医药卫生—人体生理学]