【作者】 徐静；

【导师】 牛膺筠；

【作者基本信息】 青岛大学 ， 眼科学， 2006， 硕士

【摘要】 【目的】 究热休克预处理对HSP27在视网膜色素上皮细胞中表达的影响,探讨热休克预处理对视网膜色素上皮细胞氧化损伤的保护作用及其机制。 【方法】 将培养的RPE细胞随机分成正常对照组、热休克预处理组、H₂O₂损伤组及热休克预处理加H₂O₂损伤组,并将第二、四组细胞置于42℃水浴中孵育20分钟进行热休克处理制造热休克预处理模型,再放入37℃ 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。后两组细胞600μmol·L-1H₂O₂·L-1H₂O₂处理造成RPE细胞氧化损伤模型;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H₂O₂损伤前后RPE细胞的凋亡情况。 【结果】 正常情况下RPE细胞中仅有微量HSP27表达,表达位于RPE细胞浆内,热休克预处理后RPE细胞HSP27表达出现不同程度增高,热休克预处理后2h细胞中HSP27表达开始升高,4h及8h浓度达到高峰,12h开始下降,至24h表达以接近正常水平;H₂O₂损伤组细胞经氧化损伤后RPE细胞死亡率明显上升,损伤形式主要表现为凋亡,单纯热休克预处理不引起细胞调亡,热休克预处理加H₂O₂损伤组同H₂O₂损伤组相比细胞损伤较轻,且RPE细胞凋亡的减轻程度与细胞中HSP27表达高低呈现出一定的相关性,于热休克预处理后4h及8hRPE细胞凋亡率降低最为显著。 【结论】 热休克预处理可诱导RPE细胞HSP27的高表达并能明显减轻RPE细胞的H₂O₂损伤,其机制可能主要是通过高表达的HSP27保护RPE细胞抑制其凋亡来实现的。更多还原

【Abstract】 [Objective] To study the effect of heat shock pretreatment on the express of HSP27 in retinal pigment epithelium cells and the effect on H₂O₂-induced apoptosis of retinal pigment epithelium （RPE）cells.[Methods]The cultivate retinal pigment epithelium cells were divided into 4 groups: control group,H₂O₂ injury group, heat shock pretreatment group and H₂O₂ injury after heat shock pretreatment group;cells Group 2 and 4 are culture in 42℃ water for 20min,then set in the incubator, after diffent time（0h, 2h,4h,8h,12h,24h） to break off culture and used in experiment;cells Group 3 and 4 are exposed to 600μmol·L-1 H₂O₂ to induce retina pigment epithelium cells oxidation injury;The percentage of apoptotic cells were analyzed by Flow Cytometry;heat shock proteins were detected by immunochemistry.[Results] There are mild express of hsp27 in retina pigment epithelium cells. After heat shock pretreatment the express of hsp27 are enhancement in different degree. After heat shock pretreatment 2 hours the express begin to heighten, after 4h and 8h the express reach to the peak ,and after 12h and 24h the express were begin to decline, the apoptosis of Group 3 are obviosly ,the apoptosis of Group 4 are descent in different degree. Heat shock pretreatment can inhibit the H₂O₂-induced apoptosis of RPE cells obviously.[Conclusion] Heat shock pretreatment can induce the overexpression of hsp27 and against the apoptosis induced by H₂O₂. the main mechanism perhaps was via induce the overexpression of hsp27 to protect the RPE cells and inhibit the apoptosis.更多还原