miR-24通过SIRT1调控晶状体上皮细胞凋亡的研究

目的探讨氧化应激情况下miR-24对晶状体细胞凋亡的调控。方法采用实时定量PCR检测40例白内障患者晶状体上皮组织及临近晶状体上皮组织中miR-24的表达水平,并在氧化应激情况下检测miR-24的表达变化。通过miR-24 mimics、miR-24 inhibitor转染晶状体上皮细胞SAR01/04以过表达和敲低miR-24,利用pcDNA3.1-SIRT1转染SAR01/04以过表达SIRT1,FITC/PI流式细胞术检测晶状体细胞凋亡情况,Western blot检测凋亡相关蛋白Bcl-2、Bax表达情况,CCK-8检测细胞活性状态。结果miR-24在白内障晶状体上皮组织中的表达高于临近组织,氧化应激情况下促进miR-24表达增高,差异有统计学意义(P<0.05);氧化应激情况下,敲低miR-24抑制晶状体上皮细胞的凋亡;过表达或敲低miR-24可以分别降低或促进SIRT1的表达;过表达miR-24和过表达SIRT1后抑制了晶状体细胞的凋亡。结论氧化应激促进晶状体上皮细胞miR-24表达上调,miR-24通过下调SIRT1促进晶状体上皮细胞的凋亡,为白内障的靶向治疗提供一定的策略。

miR-24 regulates apoptosis via suppressing SIRT1 in lens epithelial cells

Objective To investigate the regulation of miR-24 on apoptosis of lens cells under oxidative stress.Methods The expression level of miR-24 was detected by real-time PCR in cataract lens epithelial of 40 patients,and the change of miR-24 expression was detected under oxidative stress.SAR01/04 was transfected by miR-24 mimics and miR-24 inhibitor for over-expression or knockdown of miR-24,SAR01/04 was transfected by pcDNA3.1-SIRT1 for overexpressing SIRT1,apoptosis of lens cells was detected by FITC/PI flow cytometry,Bcl-2 and Bax expression of apoptosis related proteins were detected by Western blot,and cell activity was detected by CCK-8.Results The expression of miR-24 in the lens epithelial tissue of cataract was higher than that in the adjacent tissues,and the expression of miR-24 was significantly increased under oxidative stress(P<0.05);under oxidative stress,knockdown of miR-24 inhibited the apoptosis of lens epithelial cells;overexpression or knockdown of miR-24 could reduce or promote the expression of SIRT1;overexpression of miR-24 and SIRT1 could inhibit the apoptosis of lens epithelial cells.Conclusion Oxidative stress promotes the up-regulation of miR-24 expression in lens epithelial cells.miR-24 promotes the apoptosis of lens epithelial cells by down-regulating SIRT1,which provides certain strategies for the targeted treatment of cataract.