热休克蛋白参与NF-κB介导的H2O2预处理的细胞保护作用

目的探讨热休克蛋白(HSP)是否参与核因子-κB(NF-κB)介导的H2O2预处理的细胞保护作用。方法应用碘化丙啶(PI)染色流式细胞技术(FCM)检测细胞凋亡率,免疫印迹法(Westernblot)测定NF-κB的表达水平,免疫细胞化学染色法测定NF-κB的核转移。结果100μmol/LH2O2预处理PC12细胞90min可显著地抑制300μmol/LH2O2引起的细胞凋亡,并可明显地上调PC12细胞NF-κB的表达及促进NF-κB的核转移,同时也能上调HSP70和HSP90的表达。NF-κB的抑制剂TPCK阻断NF-κB表达,并显著地阻断H2O2预处理的细胞保护作用及HSP70和HSP90的表达。结论HSP70和HSP90参与NF-κB介导的H2O2预处理的适应性细胞保护作用。

Heat shock proteins were involved in the adaptive cytoprotection of H2O2 preconditioning mediated by nuclear factor-κB

Objective To investigate whether heat shock proteins (HSPs) were involved in the cytoprotection of H2O2 preconditioning mediated by Nuclear factor-κB (NF-κB). Methods The percent of apoptotic cells was measured by propidium iodide stain flow cytometry (FCM), the level of NF-κB expression was detected by Western blot assay, and the nuclear translocation of NF-κB p65 was assessed by immuno-cytochemical staining. Results The preconditioning of H2O2 at 100 μM markedly reduced PC 12 cells apoptosis induced by 300 μmol / L H2O2, and significantly enhanced expression and nuclear translocation of NF-κB p65. Furthermore, 100 μmol / L H2O2 preconditioning also up-regulated expression of HSP70 and HSP90. N-tosyl-L-phenylalanine chloromethyl ketone (TPCK), an inhibitor of NF-κB blocked the overexpression of NF-κB, HSP70 and HSP90 induced by H2O2 preconditioning, accompanied with attenuation of H2O2 preconditioning-induced cytoprotectioin. Conclusion HSP70 and HSP90 participated in the adaptive cytoprotection of H2O2 preconditioning mediated by NF-κB pathway.