硫化氢对抗过氧化氢对PC12细胞的损伤作用

目的探讨硫化氢(hydrogen sulfide,H2S)对抗过氧化氢(hydrogen peroxide,H2O2)对PC12细胞的损伤作用及有关机制。方法应用H2O2在PC12细胞建立氧化应激损伤的实验模型;应用甲氮甲唑蓝(MTT)法检测细胞存活率,碘化丙啶(PI)染色流式细胞技术(FCM)检测细胞凋亡率,罗丹明123(Rhodamine 123,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。应用硫化氢钠(sodium hydrosulfide,NaHS)作为H2S的供体。结果200μmol和400μmolH2O2作用PC12细胞24h均使细胞的存活率明显降低及凋亡率显著增加,200μmolH2O2引起PC12细胞的MMP明显降低及ROS生成显著增多。当NaHS与H2O2(200或400μmol/L)共同作用于PC12细胞时,NaHS(100～400μmol/L)浓度依赖性的阻断H2O2引起PC12细胞的存活率降低及细胞凋亡率增加。400μmolNaHS明显地阻断200μmolH2O2引起PC12细胞的MMP降低及ROS增多。结论H2S能明显地保护PC12细胞对抗H2O2引起的损伤,阻断MMP降低及ROS生成可能是H2S的细胞保护机制之一。

Hydrogen sulfide protects PC 12 cells against H2O2-induced damage

Objective To explore the protection of hydrogen sulfide (H2S) against hydrogen peroxide (H2O2)-induced PC12 cell damage and the mechanisms underlying. Methods PC12 cells were used to set up the oxidative stress injury experiment model by using H2O2. The viability of PC12 cells were measured by MTT assay. The percentage of apoptotic cells was assessed by propidium iodide stain flow cytometry (FCM). The mitochondrial membrane potential (MMP) was analyzed by rhodamine 123 stain FCM. The level of reactive oxygen species (ROS) in PC12 cells was tested by dihydrohodamine 123 stain FCM. Sodium hydrosulfide (NaHS) was used as H2S donor. Results H2O2 at 200 μmol and 400 μmol respectively reduced the cell viability and increased the apoptotic percentage in PC12 cells. H2O2 at 200 μmol induced the loss of MMP and overproduction of ROS in PC12 cells. However, when PC12 cells were treated by both NaHS (100 μmol/L to 400 μmol/L) and H2O2(200 μmol/L to 400 μmol/L) for 24 h, NaHS dose-dependently blocked the inhibition of cell viability and the increase in apoptotic percentage induced by H2O2. 400 μmol NaHS significantly inhibited the loss of MMP and overproduction of ROS induced by H2O2 at 200 μmol. Conclusion H2S can obviously protect PC12 cells against H2O2-induced damage, which is associated with the inhibition of NaHS on the loss of MMP and overproduction of ROS induced by H2O2.