热休克反应对氧化应激所致nucleolin裂解的影响  

Effect of heat shock response on the cleavage of nucleolin induced by oxidative stress

作　　者：王慷慨[1] 蒋磊[1] 易宇欣[1] 刘可[1] 鄂顺梅[1] 唐道林[1] 王建设[1] 石永忠[1] 王秋鹏[1] 肖献忠[1] 

机构地区：[1]中南大学湘雅医学院病理生理学教研室,长沙410078

出　　处：《中南大学学报（医学版）》2004年第5期504-508,共5页Journal of Central South University ：Medical Science

基　　金：国家自然科学基金 (30 30 0 1 77;30 2 70 533) ;国家 973重点项目 (G2 0 0 0 0 5690 8);教育部博士点专项基金 (2 0 0 2 0 5330 32 )

摘　　要：观察热休克反应对氧化应激 (0 .5mmol/LH2 O2 )诱导细胞凋亡发生时nucleolin(C2 3)裂解的影响并探讨其机制。方法 :采用caspase 3活性定量分析及免疫印迹技术检测氧化应激诱导的原代心肌细胞、RAW 2 6 4 .7巨噬细胞及K5 6 2白血病细胞凋亡及C2 3的裂解 ;通过热休克反应观察热休克反应对C2 3裂解的影响。结果 :0 .5mmol/LH2 O2处理细胞 2hcaspase 3活性显著升高 ,12h达高峰。免疫印迹结果显示上述各种细胞的C2 3蛋白均发生了裂解 (有 80kD裂解片断出现 ) ,而且最早出现裂解片段的时间都在H2 O2 处理 30min到 1h左右 ;同时热休克反应通过诱导HSP70 ,HSP2 5等应激蛋白表达而显著减少氧化应激所致C2 3的裂解。结论 :氧化应激诱导凋亡发生的同时也引起C2 3的裂解 ;热休克反应显著抑制氧化应激所致C2 3的裂解 。Objective To observe the cleavage of nucleolin (C23) during apoptosis induced by oxidative stress and to clarify the effect of heat shock response (HSR) on the cleavage of nucleolin and its possible molecular mechanism. Methods We added 0.5 mmol/L peroxide hydrogen (H 2O 2) into cultured cells to mimic oxidative stress. Apoptosis and cleavage of C23 were detected using caspase-3 colorimetric assay and Western blotting respectively. HSR was performed to observe the effect of HSR on cleavage of C23 induced by oxidative stress, and over-expressions of HSP70 and HSP25 were detected by Western blotting.Results Activity of caspase-3 increased significantly after 2 hours of 0.5 mmol/L H 2O 2 treatment, and reached the peak after 12 hours. The cleavage of C23 appeared 30 minutes to 1 hour after the treatment of H 2O 2 as indicated by a cleaved fragmentation of 80 kD, which was significantly inhibited by HSR. Moreover, HSR could induce HSP70 and HSP25 over-expressions. Conclusion Oxidative stress can induce the activation of caspase -3, cleavage of C23, and apoptosis. HSR can significantly inhibit the cleavage of C23 induced by oxidative stress, which is related to the over-expressions of HSP70, HSP25, and other stress proteins.

关 键 词：热休克反应 氧化应激 诱导 CASPASE-3 观察 表达 影响 细胞 H2O2 片段 

分 类 号：R363[医药卫生—病理学]