CAJ | 학술논문

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、H2 O2损伤组及热休克预处理加H2 O2损伤组，将热休克预处理、热休克预处理加H2 O2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％CO2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。 H2 O2损伤组及热休克预处理加H2 O2损伤组细胞600μmol· L－1 H2 O2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H2 O2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，H2 O2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表现出对H2 O2损伤的耐受性，与H2 O2损伤组比较，预处理后4h加H2 O2损伤组及8h加H2 O2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H2O2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。
목적：연구열휴극예처리대열휴극단백27（ HSP27）재시망막색소상피세포（ RPE）중표체적영향，탐토열휴극예처리대RPE양화손상적보호작용급기궤제。방법배양적RPE수궤분성정상대조조、열휴극예처리조、H2 O2손상조급열휴극예처리가H2 O2손상조，장열휴극예처리、열휴극예처리가H2 O2손상조세포치우42℃수욕중부육20min진행열휴극처리，우37℃체적분수5％CO2세포배양상중계속배양，분별우0h，2h，4h，8h，12h，24h후종지배양，용우실험。 H2 O2손상조급열휴극예처리가H2 O2손상조세포600μmol· L－1 H2 O2처리조성RPE양화손상모형；응용면역세포화학법검측열휴극예처리전후세포HSP27적표체；류식세포술검측H2 O2손상전후RPE적조망정황。결과열휴극예처리후RPE HSP27표체출현불동정도증고，여대조조비교，예처리후4h급8h조표체명현증강，Q치분별위9．9133，11．6842，P＜0．01；류식세포의분석검측결과현시，H2 O2가이현저강저세포적활성，여대조조비교，Q치위51．7561，P＜0．01。열휴극예처리후적RPE표현출대H2 O2손상적내수성，여H2 O2손상조비교，예처리후4h가H2 O2손상조급8h가H2 O2손상조RPE조망솔명현강저，Q치분별위24．1271，25．5848，P＜0．01，차정도여세포중HSP27표체고저정일정적상관성（r＝－0．9856，P＜0．01）。결론열휴극예처리가유도RPE HSP27적고표체병능명현감경RPE적H2O2손상，기궤제가능주요시통과고표체적HSP27보호RPE억제기조망래실현。
[ 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.