MicroRNA-133对心肌细胞内钙的调节机制  

Regulatory mechanism of microRNA-133 on intracellular calcium metablism in rat cardiomyocytes

作　　者：沐晓芹[1] 张莹[1] 梁海海[1] 李超[1] 陈艳艳[1] 吕延杰[1] 杨宝峰[1] 

机构地区：[1]哈尔滨医科大学药理学教研室黑龙江省生物医药重点实验室省部共建国家重点实验室培育基地,黑龙江哈尔滨150081

出　　处：《哈尔滨医科大学学报》2009年第4期328-332,共5页Journal of Harbin Medical University

基　　金：2006年度国家博士点基金项目(20060226019)

摘　　要：目的研究微小RNA133(microRNA-133,miRNA-133)对乳鼠心肌细胞内钙浓度的调节机制。方法利用lipofectamine2000将miRNA-133转染到原代培养的乳鼠心肌细胞中,采用细胞免疫荧光染色方法检测miRNA-133对L型钙通道蛋白表达的影响,并采用激光共聚焦显微镜检测miRNA-133对心肌细胞内钙浓度的影响。结果与对照组相比,转染miRNA-133的乳鼠心肌细胞钙通道蛋白的荧光强度显著降低,细胞内钙荧光强度也明显降低(P<0.05),转染AMO133(miRNA-133的反义链,可特异性阻断miRNA-133)的心肌细胞钙通道蛋白的荧光强度显著增强,细胞内钙荧光强度也明显增加(P<0.05),miRNA-133和AMO133共转染的心肌细胞钙通道蛋白的荧光强度和细胞内钙荧光强度与对照组相比无明显变化。结论MiRNA-133通过抑制L型钙通道蛋白的表达而降低心肌细胞内的钙浓度。Objective To investigate the regulation of [ Ca^2＋ ]i by microRNA-133 （miRNA-133） and underlying mechanism in rat neonatal cardiomyocytes. Methods MiRNA-133 was transfected with lipofectamine2000 into primary cultured rat neonatal cardiomyocytes, then the expression of L-type Ca^2＋ channel and [ Ca^2＋ ] i were detected with immunofluorescence technique and laser confoeal microscopy. Re＇suits The expression of L-type Ca^2＋ channel and the intracellular fluorescence intensity （ [ Ca^＋＋] i） of the cardiomyocytes in miRNA-133 group were lower than those in the control group（P 〈 0. 05） ;The fluorescence intensity of L-type Ca^2＋ channel and the intracellular fluorescence intensity of cardiomyocytes in AMO-133 group were higher than those in control group（P 〈 0. 05 ）. However, there was no significant difference in the expression and the intracellular fluorescence intensity of L-type Ca^2＋ channel between the cotransfection group （miRNA-133 ＋ AMO-133 ） and control group （P 〉 0. 05 .）. Conclusion MiRNA-133 decreased [ Ca^2＋ ]i by reducing the expression of L-type Ca^2＋ channel.

关 键 词：miRNA-133 心肌细胞 细胞钙浓度 L型钙通道 

分 类 号：R542.2[医药卫生—心血管疾病] R135.2[医药卫生—内科学]