【摘要】 目的:观察重组腺病毒pAd-CMV5-DEST-hogg1转染人视网膜色素上皮细胞ARPE-19后,高表达hOGG1对视网膜色素上皮细胞在H2O2诱导的氧化损伤中的作用。方法:鉴定和包装重组腺病毒pAd-CMV5-DEST-hogg1和腺病毒载体对照p Ad/CMV/V5-GW/lac Z,并分别转染ARPE-19细胞。H2O2(100μmol·L-1,2 h)干预转染重组腺病毒的ARPE-19细胞(RPE-Ad-hogg1)、转染腺病毒载体对照的ARPE-19细胞(RPE-Ad-lac Z)以及未转染病毒的ARPE-19细胞。蛋白免疫印迹法检测3组细胞hOGG1的表达量;流式细胞仪检测细胞内ROS的生成量;免疫细胞化学方法检测氧化损伤标志性终产物8-羟基鸟嘌呤(8-oxod G)的形成量。结果:重组腺病毒p Ad-CMV5-DEST-hogg1包装成功,并在ARPE-19细胞中表达;高表达h OGG1的视网膜色素上皮细胞较两对照组细胞显示出更少的ROS生成量(P<0.05)、更低的8-oxod G氧化损伤程度(P<0.01)。结论:视网膜色素上皮细胞hOGGl蛋白高表达,可提高ARPE-19细胞碱基水平的修复能力。更多还原

【Abstract】 Objective: To study the effect of hOGG1 on human retinal pigment epithelial cells in H2O2 induced oxidative stress,by observing p Ad-CMV5-DEST-hogg1 transfecting APRE-19. Methods: Construct recombinant adenovirus pAd-CMV5-DEST-hogg1,control adenovirus vector p Ad / CMV / V5-GW / lac Z,and transfect them into ARPE-19 cells( RPE-Ad-hogg1,RPE-Ad-lac Z) respectively. The two cell lines,together with a normal ARPE-19 cell strain,were treated by H2O2( 100 μmol·L-1,2 h). The oxygen free radicals( ROS) production was detected by Flow Cytometry( FCM). Immunohistochemisty assessed 8-oxoguanine( 8-oxod G),which is the oxidative damage end products. Results: The construction and packaging of recombinant adenovirus-hogg1 vector were successful,and it could be expressed in ARPE-19 cells. ARPE-19 cells with high h OGG1 expression had a lower ROS and 8-oxod G production( P < 0. 05) and lower degree of oxidational damage on 8-oxod G. Conclusion: A high expression of h OGG1 in ARPE-19 cells can reduce the degree of oxidative stress induced by H2O2.更多还原