摘要
【目的】探讨硫化氢(H2S)对抗二氯化钴(CoCl2)诱导PC12细胞凋亡的作用及其初步机制。【方法】应用化学性缺氧模拟剂CoCl2在PC12细胞建立化学性缺氧损伤模型;以硫氢化钠(NaHS)作为H2S的供体;应用CCK-8比色法检测细胞存活率;碘化丙啶(PI)染色流式细胞技术(FCM)检测细胞凋亡率;Hoechst33258染色检测细胞凋亡的形态学变化;罗丹明123(Rh123)染色荧光显微镜照相检测细胞线粒体膜电位(MMP);2',7'-二氯荧光黄双乙酸盐(DCFH-DA)染色荧光显微镜照相检测细胞内的活性氧(ROS)水平。【结果】CoCl2引起PC12细胞的存活率显著降低及凋亡率明显增加,同时引起PC12细胞的MMP明显降低及ROS生成显著增加。在600μmol/L CoCl2处理PC12细胞前,应用100～400μmol/L NaHS预处理30min,呈浓度依赖性地阻断CoCl2引起PC12细胞的存活率降低;200μmol/L、400μmol/L NaHS预处理能显著地降低600μmol/L CoCl2引起PC12细胞的凋亡率增加并阻断CoCl2引起的MMP降低及ROS升高。【结论】H2S具有神经细胞保护作用,能明显地对抗CoCl2诱导的PC12细胞凋亡,此作用可能与其减少ROS生成,提高MMP有关。
【Objective】 To explore the cytoprotection of hydrogen sulfide （H2S） against cobalt chloride （CoCl2）-induced apoptosis in PC12 cells and the underlying mechanisms.【Methods】 CoCl2 （a chemical hypoxia-mimetic agent） was used to establish the chemical hypoxia-induced PC12 cell injuries model.Sodium hydrosulfide （NaHS） was used as a H2S donor.The viability of PC12 cells was measured by CCK-8 assay.The percentage of apoptotic cells was assessed by propidium iodide stain flow cytometry （FCM）.The morphological change of apoptotic cells was tested by using the chromatin dye Hoechst 33258.The mitochondrial membrane potential （MMP） was analyzed by rhodamine 123 staining and photofluorography.The level of reactive oxygen species （ROS） in PC12 cells was measured by DCFH-DA staining and photofluorography.【Results】 CoCl2 induced a decrease in cell viability and an increase in percentage of apoptosis in PC12 cells along with dissipation of MMP as well as overproduction of ROS.When PC12 cells were treated with NaHS 30 min before CoCl2 treatment a decrease in viability of PC12 cells induced by 600 μmol / L CoCl2 was concentration-dependently blocked by NaHS （100,200,and 400 μmol / L）.Pretreatment with NaHS at 200 and 400 μmol / L obviously reduced the apopototic percentage of PC12 cells induced by 600 μmol / L CoCl2 and inhibited the dissipation of MMP and overproduction of ROS.【Conclusion】 H2S protected PC12 cells against CoCl2-induced apoptosis,which may be associated with the inhibition of H2S on the dissipation of MMP and overproduction of ROS induced by CoCl2.
出处
《中山大学学报（医学科学版）》
CAS
CSCD
北大核心
2010年第1期79-84,共6页
Journal of Sun Yat-Sen University:Medical Sciences
基金
广东省科技计划项目(2008B080703053)
关键词
硫化氢
二氯化钴
缺氧
PC12细胞
凋亡
hydrogen sulfide
cobalt chloride
hypoxia
PC12 cells
apoptosis
