摘要
目的探讨氧化损伤对体外培养的人视网膜色素上皮(RPE)细胞金属蛋白酶(MMP-2,9)及其组织抑制剂(TIMP-1)表达的影响。方法将不同浓度的H2O2处理传代的RPE细胞,检测活力后,采用明胶酶谱法和RT-PCR技术定量分析RPE细胞中MMP-2,9的蛋白水平和mRNA含量。采用RT-PCR和ELISA分别测定TIMP-1的mRNA表达和培养液中的浓度。结果不同浓度H2O2对RPE细胞增殖没有影响,MMP-2均呈高表达,但mRNA和蛋白均没有变化;MMP-9表达相对弱,且随着H2O2浓度的升高而增强,ELISA法检测不同浓度H2O2处理的RPE细胞培养液中TIMP-1浓度无显著差别。结论不同H2O2浓度下MMP-2、TIMP表达无明显差异,MMP-9随着H2O2浓度增加表达增强,表明糖尿病视网膜病变中存在的氧化损伤导致MMP-9表达升高。
O bjective To investigate the expression of matrix metalloproteinase-2,9(MMP -2,9)and tissue inhibitors of metalloproteinase(TIMP-1)in cultured huma n retinal pigment epithelial(RPE)cell under oxidative stress.Methods Human ARPE-19were incubated in media containing0,10,50and100μmol H 2 O 2 .The cells viability was evaluated by MTT assay,and the activity of MMPs was assessed by gelatin zymography.The levels of mRNA encoding MMP and TIMP- 1were determined by RT-PCR,and TIMP-1level in media was detected by ELISA. Results Varied concentrations of H 2 O 2 did not affect proliferation of RPE cells.MMP-2was strongly expressed in all samples,and MMP-9was weakly expressed but gradually enhanced with increa se of H 2 O 2 concentrations.There was no significant difference among the levels of TIM P-1of all samples by ELISA.Conclusion There was no obvious difference of MMP -2and TIMP expressions in various H 2 O 2 concentrations groups.MMP-9may be involved in the oxidative stress and pa thogenesis of diabetic retinopathy.
出处
《眼科研究》
CSCD
北大核心
2004年第1期40-43,共4页
Chinese Ophthalmic Research
