缺氧条件下Hsp90抑制剂17-AAG对RPE细胞整联蛋白连接激酶表达的影响  

Effect of 17-AAG,a Hsp90 inhibitor,on the expression of ILK in RPE cells under hypoxic condition

作　　者：王业青[1] 王健[1] 吕勇[1] 董丽[1] 穆华[1] 曹文萍[1] 

机构地区：[1]哈尔滨医科大学附属第一医院眼科,中国黑龙江省哈尔滨市150001

出　　处：《国际眼科杂志》2014年第5期811-814,共4页International Eye Science

基　　金：黑龙江省教育厅科学技术研究项目(No.12531389)~~

摘　　要：目的:研究热休克蛋白(heat shock protein 90,Hsp90)抑制剂17-AAG对缺氧诱导的视网膜色素上皮(retinal pigment epithelial,RPE)细胞整联蛋白连接激酶(integrin-linked kinase,ILK)表达的影响。方法:利用200μmol/L氯化钴(cobalt chloride,CoCl2)建立体外RPE细胞的化学缺氧模型。不同浓度的Hsp90抑制剂17-AAG预处理RPE细胞1h后给予12h缺氧处理。实验分为缺氧对照组、二甲基亚砜(DMSO)对照组和17-AAG预处理组。其中17-AAG预处理组根据浓度不同又分为6组:0.01,0.10,0.50,1.00,5.00,10.00μmol/L。应用RT-PCR和Western blot方法检测ILK表达变化情况。结果:缺氧对照组、DMSO对照组和17-AAG预处理组ILK mRNA与内参β-actin mRNA光密度比值分别为1.32±0.04,1.29±0.03,0.93±0.06,0.70±0.05,0.53±0.03,0.44±0.04,0.32±0.04,0.30±0.03;ILK蛋白与内参β-actin蛋白光密度比值分别为2.16±0.04,2.13±0.04,1.65±0.04,1.13±0.05,0.74±0.03,0.41±0.06,0.35±0.04,0.35±0.03。与缺氧对照组比较,17-AAG预处理组ILK mRNA和蛋白的表达明显降低(P<0.05),呈浓度依赖性。结论:Hsp90抑制剂17-AAG能够降低缺氧条件下RPE细胞ILK的表达。AIM： To investigate the effect of 17-allylamio-17-demethoxygeldanamycin （ 17 - AAG ） , an heat shock protein 90（Hsp90） inhibitor, on the expression of integrin-linked kinase （ ILK） induced by hypoxia in retinal pigment epithelium （ RPE） cells. METHODS： RPE cells were cultured with 200μmol/L cobalt chloride （ CoCl2 ） for 12 hours to imitate chemical hypoxia. Pretreatment of 17-AAG was 1 hour prior to hypoxic insult. Experiment was divided into hypoxic control group, Dimethyl sulfoxide （ DMSO） control group and 17-AAG pretreatment group （0. 01, 0. 10, 0. 50, 1. 00, 5. 00 and 10. 00μmol/L ）. RT - PCR and Western blot analysis were used to examine the expression of ILK in cultured RPE cells. RESULTS：The density ratios of ILK mRNA and β-actin mRNA of hypoxic control group, DMSO control group and 17-AAG pretreatment groups were 1. 32±0. 04,1. 29±0. 03, 0. 93±0. 06,0. 70±0. 05,0. 53±0. 03,0. 44±0. 04,0. 32±0. 04,0. 30±0. 03;and the density ratios of ILK protein and β-actin protein＆amp;nbsp;were 2. 16±0. 04, 2. 13±0. 04, 1. 65±0. 04, 1. 13±0. 05, 0. 74±0. 03, 0. 41± 0. 06, 0. 35± 0. 04, 0. 35± 0. 03. The ILK expression in 17-AAG pretreated groups were inhibited compared to hypoxic control group （P 〈 0. 05）, and the decrease was in concentration-dependent manner. CONCLUSION：Hsp90 inhibitor 17-AAG was effective to inhibit the expression of ILK induced by hypoxia in RPE cells.

关 键 词：整联蛋白连接激酶 视网膜色素上皮细胞 缺氧 热休克蛋白90 17-丙烯胺基-17-去甲氧基格尔德霉素 

分 类 号：R773.4[医药卫生—眼科]