过氧化氢对人视网膜色素上皮细胞ARPE-19中20S蛋白酶体表达的上调作用

Up-regulation effect of hydrogen peroxide on expression of 20S proteasome in ARPE-19

目的:探讨过氧化氢诱导的氧化应激对人视网膜色素上皮细胞ARPE-19中20S蛋白酶体表达的影响。方法:分别选择0,100,300μmol/L过氧化氢作用ARPE-19细胞4h后收集样本,Western blot方法检测细胞中20S蛋白酶体表达水平,细胞免疫荧光方法检测20S蛋白酶体在细胞中的分布。结果:100和300μmol/L过氧化氢溶液处理的细胞中20S蛋白酶体的表达和对照组相比明显增加(100μmol/L过氧化氢处理组vs0和300μmol/L过氧化氢处理组,P<0.05),并呈现剂量依赖性。正常培养情况下,细胞内20S蛋白酶体在核周微量表达,100和300μmol/L过氧化氢溶液处理的细胞内20S蛋白酶体和对照组相比明显增加,其分布集中在核内及核膜周围。结论:过氧化氢对ARPE-19的20S蛋白酶体表达有上调作用。

AIM： To investigate the effect of hydrogen peroxide on 20S proteasome expression induced by oxidative stress in ARPE-19 cells in vitro. METHODS： ARPE-19 were treated with 0, 100 or 300μmol/L hydrogen peroxide for 4 hours, respectively, then the expression and distribution of 20S proteasome in cells were detected by Western Blot and immunofluorescence staining assay respectively. RESULTS： 20S proteasome increased significantly in ARPE-19 when it treated by hydrogen peroxide at 100 and 300μmol/L （100μmol/L hydrogen peroxide vs 0, 300μmol/ L hydrogen peroxide , P 〈 0.05）, and the up-regulation change was dose-dependent. Intracellular distribution of 20S proteasome in ARPE-19 cultured under normal circumstances was around the nuclear membrane. 20S proteasome expression in ARPE-19 increased after treated by hydrogen peroxide at 100 and 3001jmol/L, mainly in the nucleus and around the nuclear membrane. CONCLUSION： 20S proteasome is up-regulated inARPE-19 treated by hydrogen peroxide. Ubiquitin- proteasorne pathway is induced in ARPE-19 by oxidative stress in vitro.