线粒体靶向性肽SS-31对H2O2诱导人晶状体上皮细胞凋亡的保护作用

doi:10.6040/j.issn.1671-7554.0.2014.195

摘要/Abstract

摘要： 目的研究线粒体靶向性肽SS-31在H₂O₂诱导的人晶状体上皮细胞凋亡中的保护作用及其分子机制.方法体外培养人晶状体上皮细胞系SRA01/04,选取相同代次的SRA01/04细胞进行实验,应用100 μmol/L H₂O₂ 构建凋亡模型,随机分为正常对照组、H₂O₂处理组、3个不同浓度的SS-31预处理组.应用MTT法检测各组细胞存活率;流式细胞术检测各组细胞凋亡率;Western blotting检测各组凋亡蛋白Bcl-2、Bax、Cleaved-Caspase-3的表达.结果 H₂O₂处理24 h后,细胞存活率降低,凋亡率升高,促凋亡蛋白Bax的表达升高,抗凋亡蛋白Bcl-2的表达降低,Bax/Bcl-2比率升高,Cleaved-Caspase-3的表达升高.SS-31预处理能明显提高细胞的存活率,且与浓度呈正相关.同时,SS-31还能减少细胞的凋亡率,下调Bax的表达,上调Bcl-2的表达,减少Cleaved-Caspase-3的激活,从而发挥抗凋亡作用.结论 SS-31能在H₂O₂诱导的人晶状体上皮细胞凋亡中有效地发挥保护作用,但能否应用于白内障治疗有待深入研究.

关键词: SS-31, 人晶状体上皮细胞, 氧化应激, 凋亡, 线粒体靶向性肽

Abstract: Objective To study the protective effect of SS-31 against H₂O₂-induced apoptosis in human lens epithelial cells and the possible mechanism. Methods SRA01/04 cells, an immortal human lens epithelial cell line, were cultured in vitro. SRA01/04 cells at the same generation were chosen and 100 μmol/L H₂O₂ was used to build the model of apoptosis. The cells were randomly allocated into five groups: the control group, the group treated by H₂O₂, and three groups pretreated by different concentrations of SS-31. The cell viabilities were monitored by using 4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide(MTT) assay. The cell apoptotic rates were detected with Annexin V-Propidium iodide double-staining flow cytometry. The expressions of Bcl-2, Bax and Cleaved-Caspase-3 were tested by using Western blotting. Results After exposure of 100 μmol/L H₂O₂ for 24 h, the cell viabilities decreased, cell apoptotic rates increased, expression of pro-apoptotic protein Bax increased, expression of anti-apoptotic protein Bcl-2 decreased, and expression of Cleaved-Caspase-3 increased. Pretreatment of SS-31 at different concentrations improved cell viabilities in a dose-dependent manner. Meanwhile, SS-31 plays an anti-apoptotic role by decreasing cell apoptotic rate, down-regulating the expression of Bax, up-regulating the expression of Bcl-2, and suppressing the activation of Cleaved-Caspase-3. Conclusion SS-31 efficiently protects human lens epithelial cells against H₂O₂-induced apoptosis. Further studies are needed on whether SS-31 can be used in medicine treatment for cataract.

Key words: SS-31, Human lens epithelial cells, Apoptosis, Mitochondria-targeted peptide, Oxidative stress

中图分类号:

R776.1

郭长艳, 秦雪娇, 吴晓莉, 高雪, 蔡可丽. 线粒体靶向性肽SS-31对H₂O₂诱导人晶状体上皮细胞凋亡的保护作用[J]. 山东大学学报（医学版）, 2015, 53(8): 23-26.

GUO Changyan, QIN Xuejiao, WU Xiaoli, GAO Xue, CAI Keli. Protective effect of mitochondria-targeted peptide SS-31 against H₂O₂-induced apoptosis in human lens epithelial cells[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2015, 53(8): 23-26.

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线粒体靶向性肽SS-31对H₂O₂诱导人晶状体上皮细胞凋亡的保护作用

Protective effect of mitochondria-targeted peptide SS-31 against H₂O₂-induced apoptosis in human lens epithelial cells

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