摘要
目的探讨热休克蛋白90(Hsp90)在硫化氢(H2S)保护PC12细胞对抗氯化钴(CoCl2)引起的化学性缺氧损伤中的作用。方法在PC12细胞建立H2S预处理对抗CoCl2诱导PC12细胞损伤的实验模型。应用细胞计数试剂盒8(CCK-8)检测细胞存活率;Hoechst 33258染色荧光显微镜照相术检测凋亡PC12细胞的形态学改变;应用碘化丙啶(PI)染色流式细胞术检测细胞凋亡率;免疫印迹法(Western blot)检测Hsp90的表达。结果H2S的供体400μmol·L-1硫氢化钠(NaHS)可上调PC12细胞Hsp90的表达,NaHS作用3h,Hsp90表达达最高峰,作用24h时表达恢复到基础水平;NaHS也能明显地增加CoCl2引起的Hsp90的表达上调。NaHS预处理能对抗CoCl2引起的PC12细胞损伤,提高细胞存活率,降低细胞凋亡率。Hsp90抑制剂17-丙烯胺基-17-去甲氧基格尔德霉素(17-AAG)可拮抗NaHS预处理对Hsp90表达的上调作用,并明显地减弱H2S诱导的适应性细胞保护作用。结论H2S能保护PC12细胞对抗CoCl2诱导的低氧损伤作用,诱导Hsp90表达上调可能是其细胞保护机制之一。
Aim To explore the role of heat shock protein 90（Hsp90）in hydrogen sulfide-induced protection against PC12 cells injuries elicited by cobalt chloride （CoCl2）. Methods Hydrogen sulfide preconditioning against CoCl2-induced injury model was set up in PC12 cells. Cell viability was tested by using cell counter kit-8. Morphological changes in apoptotic PC12 cells were detected by Hoechst 33258 staining and photofluorography. Apoptotic rate was evaluated by propidium iodide （PI）staining and flow cytometry（FCM）. The expression of Hsp90 was evaluated by Western blot. Results Hsp90 expression was upregulated after treatment with 400 μmol·L-1 sodium hydrosulfide（ a H2S donor）,peaking at 3 h,returning to the basal level at 24 h. Furthermore,H2S preconditioning obviously en-hanced the upregulation of Hsp90 expression induced by CoCl2. H2S preconditioning markedly protected PC12 cells against injuries induced by CoCl2,increasing the viability of cells and decreasing the percentage of apoptotic cells. 17-allylamino-17-demethoxygeldanamycin（17-AAG）,an inhibitor of Hsp90,antagonized H2S preconditioning-induced Hsp90 activation and the adaptive cytoprotection. Conclusion H2S can protect PC12 cells against CoCl2-induced injuries,and upregulating the expression of Hsp90 may be one of the mechanisms underlying cytoprotection induced by H2S pre-conditioning.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2010年第1期103-107,共5页
Chinese Pharmacological Bulletin
基金
广东省科技计划资助项目(No2008B080703053
2007B080701030)
关键词
硫化氢
氯化钴
化学性缺氧
热休克蛋白
细胞保护
凋亡
PC12细胞
hydrogen sulfide cobalt chloride chemical hypoxia heat shock proteins cytoprotection apoptosis PC12 cells
