摘要
目的探讨微小RNA(miRNA)-1对大鼠心肌细胞凋亡的影响。方法应用转染的方法使培养的大鼠H9c2心肌细胞高表达miRNA-1(miRNA-1组),real-time PCR方法检测miRNA-1表达水平以确定转染成功后,MTT法检测细胞活力,流式细胞术测定细胞凋亡情况,分别采用real-time PCR和Western Blot方法检测凋亡抑制基因Bcl-2的mRNA和蛋白表达情况,并与常规条件下培养的H9c2细胞(空白对照组)和转染随机合成的miRNA阴性对照片段(阴性对照组)比较。结果与空白对照组和阴性对照组相比,转染miRNA-1 mimics后,H9c2大鼠心肌细胞miRNA-1表达水平明显升高,细胞凋亡率显著增加,细胞生存率、Bcl-2的mRNA和蛋白表达水平显著降低。结论转染的miRNA-1 mimics能够上调细胞内的miRNA-1表达水平,抑制心肌细胞增殖,促进细胞凋亡。
Objective To investigate the effect of miRNA-1 on the cardiomyocyte apoptosis in rats. Methods MicroRNA-1 mimics was transfected into the cultured H9c2 cell line （miRNA-1 group）. Cells transfected with random miR-NA fragment was used as negative control group. The cell apoptosis was evaluated by FCM assay. MTT assay was used to de-tect the cell viability. The expression level of miRNA-1 was detected by real-time PCR. The expression levels of Bcl-2 mRNA and protein were detected by real-time PCR and Western blot assay. Results Compared with normal and negative control groups, the expression level of miRNA-1 was significantly higher in H9c2 cardiomyocytes, the apoptosis rate was in-creased, the cell vitality and Bcl-2 expression level were significantly decreased after transfection of miRNA-1 mimics. Conclusion miRNA-1 mimics can up-regulate miRNA-1 level, inhibit proliferation and induce cardiomyocyte apoptosis.
出处
《天津医药》
CAS
北大核心
2014年第5期455-457,共3页
Tianjin Medical Journal
