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为探讨热休克蛋白 70对氧化应激 (0 .5mmol L过氧化氢 )所致核仁素裂解的影响及其机制 ,采用免疫印迹技术检测氧化应激诱导核仁素的裂解 ;通过热休克反应和构建热休克蛋白 70转基因细胞观察热休克反应与热休克蛋白 70对核仁素裂解的影响 ,同时采用免疫共沉淀技术检测热休克蛋白 70与核仁素之间的相互作用。结果发现 ,0 .5mmol L过氧化氢可导致多种细胞的核仁素发生裂解 (有 80kDa裂解片断出现 ) ,而且最早出现裂解片段的时间都在过氧化氢处理 30min到 1h左右 ;热休克预处理及转热休克蛋白 70后均引起热休克蛋白 70的表达明显增加 ,同时显著抑制抑制氧化应激所致核仁素裂解片段的出现 ,免疫共沉淀结果显示在氧化应激时热休克蛋白70与核仁素直接结合 ,形成复合物。以上结果提示 ,热休克反应与热休克蛋白 70可以显著抑制氧化应激所致核仁素的裂解 ,其机理与氧化应激时热休克蛋白 70与核仁素直接结合有关。
To investigate the effect and mechanism of heat shock protein 70 on nucleolysis by oxidative stress (0.5 mmol L hydrogen peroxide), oxidative stress-induced cleavage of nucleolin was detected by Western blotting. Heat shock reaction And to construct heat shock protein 70 transgenic cells to observe the effect of heat shock reaction and heat shock protein 70 on nucleolide cleavage. At the same time, the co-immunoprecipitation assay was used to detect the interaction between heat shock protein 70 and nucleolin. The results showed that 0.5 mmol L of hydrogen peroxide led to the cleavage of nucleotides (with 80 kDa cleavage fragments) in a variety of cells, and the first time the cleaved fragment appeared in the hydrogen peroxide treatment 30min to 1h; heat shock pre After heat shock protein 70 was transferred to heat shock protein 70, the expression of heat shock protein 70 was significantly increased and the inhibition of oxidative stress was also observed. The results of co-immunoprecipitation showed that heat shock protein 70 And nucleotides directly bound to form a complex. The above results suggest that heat shock reaction and heat shock protein 70 can significantly inhibit oxidative stress-induced cleavage of nucleotides, which is related to the direct binding of heat shock protein 70 to nucleolin by oxidative stress.