Survivin在过氧化氢预处理诱导的适应性细胞保护中的作用

Role of survivin in the adaptive cytoprotection induced by H_2O_2 preconditioning

目的探讨H2O2预处理对存活素(survivin)表达的影响及存活素在H2O2预处理的适应性细胞保护中的作用。方法在PC12细胞建立H2O2预处理对抗H2O2诱导细胞凋亡的实验模型,应用甲氮甲唑蓝(MTT)法检测细胞存活率,碘化丙啶(PI)染色流式细胞术检测细胞凋亡率,Hoechst染色检测细胞凋亡的形态学改变,免疫印迹法(Westernblot)测定存活素的表达水平。结果用100μmol/LH2O2预处理PC12细胞90min可明显地抑制300μmol/LH2O2作用12h后引起的细胞毒性和细胞凋亡,并可以显著地上调PC12细胞存活素的表达;JAK2抑制剂AG-490不仅可以抑制存活素的表达,而且拮抗H2O2预处理的适应性细胞保护作用。结论存活素是JAK-STAT通路的靶基因,可能在H2O2预处理诱导的适应性细胞保护机制中起着重要的作用。

Objective To investigate the influence of H2O2 preconditioning on survivin expression and the role of survivin in the adaptive cytoprotection induced by H2O2 preconditioning in PC12 cells. Methods In PC12 cells, setting up the experimental model of cytoprotection of H2O2 preconditioning against apoptosis induced by H2O2. The viability of PC12 cells was assessed by MTT assay, the percent of apoptotic cells was measured by propidium iodide stain flow cytometry （FCM）, the morphological change of apoptotic ceils was tested by using the chromatin dye Hoechst 33258, the level of survivin expression was detected by Western blot assay. Results The preconditioning of H2O2 at 100μmol for 90 min obviously depressed apoptosis and cytotoxicity induced by 300 μmol H2O2, and up-regulated survivin expression; the inhibitor of JAK2 AG-490 inhibited not only the expression of survivin, but also the adaptive cytoprotection induced by H2O2 preconditioning. Conclusion Survivin is the target gene of JAK-STAT pathway, and played an important role in the cytoprotective effect of H2O2 preconditioning.