热休克处理对视网膜色素上皮细胞氧化损伤的保护作用  

Protective Effects of Heat Shock Pretreatment on Oxidative Injury Human Retinal Pigment Epithelial Cells

作　　者：徐静[1] 陈尧[2] 牛膺筠[3] 

机构地区：[1]潍坊市人民医院眼科,山东潍坊261041 [2]潍坊市人民医院耳鼻咽喉科 [3]青岛大学医学院附属医院眼科

出　　处：《潍坊医学院学报》2015年第1期67-69,共3页Acta Academiae Medicinae Weifang

基　　金：国家自然科学基金赞助课题(课题编号:30572010);山东省卫生和计划生育委员会;山东省医药卫生科技发展计划资助课题(课题编号:2013WS0281)

摘　　要：目的研究热休克预处理对热休克蛋白27(HSP27)在视网膜色素上皮细胞(RPE)中表达的影响,探讨热休克预处理对RPE氧化损伤的保护作用及其机制。方法培养的RPE随机分成正常对照组、热休克预处理组、H_2O_2损伤组及热休克预处理加H_2O_2损伤组,将热休克预处理、热休克预处理加H_2O_2损伤组细胞置于42℃水浴中孵育20min进行热休克处理,于37℃体积分数5%CO_2细胞培养箱中继续培养,分别于0h,2h,4h,8h,12h,24h后终止培养,用于实验。H_2O_2损伤组及热休克预处理加H_2O_2损伤组细胞600μmol·L^(-1)H_2O_2处理造成RPE氧化损伤模型;应用免疫细胞化学法检测热休克预处理前后细胞HSP27的表达;流式细胞术检测H_2O_2损伤前后RPE的凋亡情况。结果热休克预处理后RPE HSP27表达出现不同程度增高,与对照组比较,预处理后4h及8h组表达明显增强,Q值分别为9.9133,11.6842,P<0.01;流式细胞仪分析检测结果显示,H_2O_2可以显著降低细胞的活性,与对照组比较,Q值为51.7561,P<0.01。热休克预处理后的RPE表现出时H_2O_2损伤的耐受性,与H_2O_2损伤组比较,预处理后4h加H_2O_2损伤组及8h加H_2O_2损伤组RPE凋亡率明显降低,Q值分别为24.1271,25.5848,P<0.01,且程度与细胞中HSP27表达高低呈一定的相关性(r=-0.9856,P<0.01)。结论热休克预处理可诱导RPE HSP27的高表达并能明显减轻RPE的H_2O_2损伤,其机制可能主要是通过高表达的HSP27保护RPE抑制其凋亡来实现。[ ABSTRACT] Objective To study the effects of heat shock pretreatment on the express of HSP 27 in retinal pigment epithelium cells and H2O2-induced apoptosis of retinal pigment epithelium （RPE）cells.Methods The cultivate retinal pigment epithelium cells were di-vided into 4 groups：control group ,H2 O2 injury group ,heat shock pretreatment group and H 2 O2 injury after heat shock pretreatment group;heat shock pretreatment group and H 2 O2 injury after heat shock pretreatment group cells were cultured in 42℃water for 20min,then were set in the incubator,after diffent time（0h,2h,4h,8h,12h,24h） to break off culture and used in experiment;H2 O2 injury group and H2 O2 injury af-ter heat shock pretreatment group cells were exposed to 600μmol· L-1 H2 O2 to induce retina pigment epithelium cells oxidation injury .Apop-tosis was assessed by annexin V-fluorescein isothiocyanate-Propidium iodium （ annexin V-FITC-PI ） staining method and immunohistochemical methods were applied to detect the expressions of HSP 27.Results After heat shock pretreatment the express of HSP 27 enhanced in different degree.Compared with the control group ,after heat shock pretreatment 4h and 8h groups the expression of HSP27 increased obviously（Q val-ues were：9.9133,11.6842,P〈0.01）.In oxidative injury group,cells viability decreased obviously.Compared with the control group（Q=51.7561,P〈0.01）,heat shock pretreatment intervention group compared with the H 2O2 injury group,H2O2 injury after heat shock pretreat-ment 4hgroup,H2O2 injury after heat shock pretreatment 8h group cell viability gradually increased （Q values were：24.1271,25.5848,P〈0. 01） and the decrease of apoptosis had high correlation with the express of HSP 27（r=-0.9856,P〈0.01）.Conclusion Heat shock pretreat-ment can induce the overexpression of hsp 27 and against the apoptosis induced by H 2 O2 .The main mechanism of protecting the RPE cells and inhibiting the apoptosis may be via inducing the overexpression of hsp 27 to.

关 键 词：视网膜色素上皮细胞 细胞凋亡 过氧化氢 热休克预处理 

分 类 号：R774.1[医药卫生—眼科]