微小RNA-34a调控高糖诱导H9c2心肌细胞凋亡的初步研究

miR-34aregulates high glucose-induced apoptosis of H9c2cells

目的探讨微小RNA-34a(miR-34a)对高糖诱导的H9c2心肌细胞凋亡的影响。方法将培养的H9c2心肌细胞随机分为3组:正常组(葡萄糖浓度5.5mmol/L)、高糖组(葡萄糖浓度33mmol/L),抑制剂组(miR-34a抑制剂浓度50nmol/L+葡萄糖33mmol/L)。采用定量PCR检测miR-34a和凋亡相关基因Bcl-2基因表达的变化,Western blot检测凋亡相关蛋白Bcl-2表达的变化;采用流式细胞术检测细胞凋亡。结果与正常组比较,高糖组心肌H9c2细胞凋亡率明显升高(P<0.05),H9c2细胞miR-34a表达显著上调(P<0.05),H9c2细胞Bcl-2mRNA和蛋白表达减少(P<0.05)。与高糖组比较,抑制剂组H9c2细胞凋亡明显下降(P<0.05),H9c2细胞Bcl-2mRNA和蛋白表达明显升高(P<0.05)。结论 miR-34a能调控高糖所致的H9c2心肌细胞凋亡,可能是通过直接抑制抗凋亡基因Bcl-2的表达而实现的。

Objective To study the effect of miR-34a on regulation of high glucose-induced apopto sis of H9c2 cells. Methods Cultured H9C2 ceils were randomly divided into normal glucose （5.5 mmol/L） group,high glucose （33 mmol/L） group and miR-34a inhibitor （50 mmol/L）＋ glucose （33mmol/L） group. Expressions of miR-34a and Bcl-2 mRNA were detected by RT-PCR. Western blot was used to monitor the changes in apopotosis-associated protein Bcl-2,respectively. Apoptosis of-H9c2 ceils was detected by flow cytometry. Results The apoptpsis level of H9c2 ceils and the expression level of miR-34a in H9c2 cells were significantly higher whereas the expression level of Bcl-2 was significantly lower in high glucose group than in normal glucose group （P~ 0.05）. The apoptpsis level of H9c2 cells was significantly lower whereas the expression level of t3cl-2 was significantly higher in miR-34a inhibitor ＋ glucose group than in high glucose group （P〈0.05）. Conelusion miR-34a regulates high glucose-induced apoptpsis of H9c2 cells by inhibiting the expression of gel-2.