Tubulin和HSP90在氧化应激预适应中的作用

Role of heat shock protein 90 and tubulin in oxidative stress preconditioning

目的:探讨热休克蛋白90和细胞骨架蛋白tubulin在氧化应激预适应中的作用。方法:应用不同浓度H2O2作用于HepG2细胞,建立HSP90不同表达量的模型后,MTT比色法检测细胞活力,Western blotting定量检测各模型中HSP90、tubulin量,激光扫描共聚焦显微镜测定该两种蛋白在对照、预适应、氧化应激、预适应后氧化应激下的分布和共定位。结果:MTT比色法结果提示预适应可提高应激状态下的细胞生存活力;Western blotting结果显示氧化应激预适应可提高细胞内HSP90和tubulin含量,减轻再次应激所致的HSP90和tubulin总量下降;激光共聚焦显示该两种蛋白有相似的细胞分布。结论:Tubulin可能协同HSP90在氧化应激预适应中起保护作用。

AIM： To find the role of heat shock protein 90 （HSP90） and tubulin in oxidative stress preconditioning in HepG2 cells. METHODS： The different doses of H202 were used to induce cell injury in HepG2 cells. MTT assay, Western blotting and confocal laser microscopy were also used. RESULTS： MTT colorimetry showed that preconditioning （50 mmol/L H2O2 ） provided a temporary resistance against subsequent oxidative stress （500 mmol/L H2O2 ）. Western blotting demonstrated that preconditioning increased the levels of HSP90 and tubulin in HepG2 cells, and lessen the declining of HSP90 and tubulin after stress. Tubulin and HSP90＇s colocalizations in cells with different doses of H2O2 were also observed under laser scanning confocal microscope. CONCLUSION ： Tubulin might play important role in oxidative stress preconditioning in HepG2 cells by combining with HSP90.