摘要
目的探讨硫化氢(hydrogen sulfide,H2S)对抗β-淀粉样蛋白(β-amylmoid)诱导PC12细胞凋亡的作用及机制。方法应用硫化氢钠(sodium hydrosulfide,NaHS)作为H2S的供体,甲氮甲唑蓝(MTT)法检测细胞存活率,碘化丙啶(PI)染色流式细胞技术(FCM)检测细胞凋亡率,Hoechst染色检测细胞凋亡的形态学变化,罗丹明123(Rhodamine123,Rh123)染色FCM检测细胞线粒体膜电位(mitochondrial membrane potential,MMP),双氢罗丹明123染色FCM检测细胞内活性氧(reactive oxygen species,ROS)的含量。结果淀粉样多肽β25-35(Aβ25-35)引起PC12细胞的存活率显著降低及凋亡率明显增大,同时引起PC12细胞的MMP明显降低及ROS生成率显著增加。当NaHS与Aβ25-35共同作用于PC12细胞时,NaHS浓度依赖性地阻断20μmol/LAβ25-35引起PC12细胞的存活率降低,100μmol/LNaHS显著地降低20μmol/LAβ25-35引起PC12细胞的凋亡率并阻断Aβ25-35引起的MMP降低及ROS升高。结论H2S的供体NaHS具有细胞保护作用,能对抗Aβ25-35引起的PC12细胞凋亡,此作用可能与其阻断MMP降低及ROS生成增多有关。
Objective To explore the cytoprotection of hydrogen sulfide （H2S） against β-amyloid-induced apoptosis in PC12 cells and the underlying mechanisms. Methods Sodium hydrosulfide （NaHS） was used as a H2S donor. The viability of PC12 ceils was measured by MTT 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 stain FCM. The level of reactive oxygen species （ROS） in PC12 cells was measured by dihydrohodamine 123 stain FCM. Results β-Amyloid peptide 25-35 （Aβ25-35） induced a decrease in viability and an increase in percentage of apoptosis of PC12 cells along with dissipation of MMP as well as well overproduction of ROS. When PC12 cells were co-treated with Naris and Aβ25-35, a decrease in viability of PC12 cells induced by 20 μmol/L Aβ25-35 was concentration-dependently blocked by Naris （50, 100 and 200 μmol/L）. NaHS at 100 μmol/L obviously reduced the apopototic percentage of PC12 cells induced by 20 μmol/L Aβ25-35 and inhibited the dissipation of MMP and overproduction of ROS. Conclusion H2S protected PC12 cells against Aβ25-35-induced apoptosis, which may be associated with the inhibition of NaHS on the dissipation of MMP and overproduction of ROS induced by Aβ25-35.
出处
《解剖学研究》
CAS
2007年第2期107-110,共4页
Anatomy Research
基金
广东省科技计划项目(2006B36004022)
