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.
出处
《哈尔滨医科大学学报》
CAS
北大核心
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)
