摘要
【目的】探讨ATP敏感性钾通道(KATP)在硫化氢(H2S)对抗β-淀粉样多肽(Aβ)诱导嗜铬细胞瘤细胞(PC12)细胞损伤中的作用。【方法】应用硫化氢钠(NaHS)作为H2S的供体,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(Rh123)染色FCM检测细胞线粒体膜电位(MMP),双氢罗丹明123染色FCM检测细胞内活性氧(ROS)的含量。【结果】20μmol/L和40μmol/LAβ25-35能明显地诱导PC12细胞凋亡,增加细胞凋亡率,并能明显地抑制PC12细胞MMP及增加细胞内ROS生成;100mol/L和200μmol/LNaHS本身不损伤PC12细胞,但能明显地抑制Aβ25-35的致细胞凋亡作用,降低PC12细胞的凋亡率,并能显著地抑制Aβ25-35引起的MMP降低及胞内ROS水平增多;在应用NaHS与Aβ25-35作用PC12细胞之前30min,应用KATP抑制剂Glybenclamide(Gly)(10μmol/L)对PC12细胞进行预处理,能部分地对抗NaHS的细胞保护作用,使PC12细胞凋亡率增加,MMP降低及ROS生成增多。【结论】NaHS能明显对抗Aβ25-35对PC12细胞的损伤作用,此细胞保护作用可能与NaHS保护PC12细胞的MMP及抑制胞内ROS生成有关;KATP通道抑制剂Gly能部分地阻断NaHS的细胞保护作用,提示KATP通道开放可能是NaHS对抗Aβ25-35损伤作用的机制之一。
[Objective] To explore the role of ATP-sensitive K^＋ channels in the protection of hydrogen sulfide （H2S） against β-amyloid-induced injury in PC12 cells. [Methods] Sodium hydrosulfide （NariS） was used as a H2S donor. 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 rbodamine 123 stain FCM. The level of reactive oxygen species （ROS） in PC12 cells was measured by dihydrohodamine 123 stain FCM. [Results] Amyloid β-peptide 25-35 （Aβ25-35） at 20 μmol/L and 40μmol/L significantly induces PC12 cells apoptosis; and inhibits MMP and induces ROS in PC12 cells. NariS at 100 μmol/L or 200 μmol/L alone did not damage PC12 cells,but obviously attenuated Aβ25-35 induced apoptosis,and blocked the dissipation of MMP and the over-production of ROS induced by Aβ25-35 The ATP-sensitive K^＋ channel （KATP） inhibitor, glybenclamide （Gly）, which was pretreated 30 min before co-administration of NariS and Aβ25-35, obviously and partly blocked the cytoprotection of NariS, increasing the percentage of apoptosis; Furthermore, Gly at 10 μmol/L partly and significantly blocked the preservation of MMP and inhibition of overproduction of ROS afforded by NariS. [Conclusion] NariS obviously protected PC12 cells against the damage induced by Aβ25-35, the cytoprotection of NariS was associated with the preservation of MMP and inhibition of ROS over-production; KATP channel inhibitor, Gly, significantly partly blocked the cytoprotection of NariS, indicating that KATP channels activation played an important role in the protection of NariS against PC12 cells damage induced by Aβ25-35.
出处
《中山大学学报（医学科学版）》
CAS
CSCD
北大核心
2008年第3期258-263,共6页
Journal of Sun Yat-Sen University:Medical Sciences
基金
广东省科技计划基金(2004B30601016)
广东省自然科学基金(5001676)
关键词
硫化氢
Β-淀粉样多肽
线粒体膜电位
活性氧
ATP敏感性钾通道
hydrogen sulfide
β-amyloid peptide
mitochondrial membrane potential
reactive oxygen species
ATP-sensitive K^＋ channels
