姜黄素对H_2O_2损伤PC12细胞的保护作用

Protective effect of curcumin on H_2O_2-induced damage in PC12 cells

目的 探讨姜黄素 (curcumin ,Cur)对氧化应激损伤PC12细胞的保护作用。方法 以H2 O2 损伤PC12细胞为氧化应激损伤的模型 ,采用甲氮甲唑蓝 (3 [4 ,5 dimethylthia zol 2 yl] 2 ,5diphenyltetrazoliumbromide ,MTT)法检测细胞增殖状况 ,碘化丙啶 (Propidiumiodide,PI)染色流式细胞术(flowcytometry ,FCM )检测细胞凋亡 ,罗丹明 12 3(Rho damine12 3,Rh12 3)染色FCM检测细胞线粒体膜电位 (mito chondrialpotentialmembrane ,△Ψm) ,双氢罗丹明 12 3(Dihy drohodamine12 3,DHR)染色FCM检测细胞内活性氧 (reac tiveoxygenspecies,ROS)的含量。 结果 2 0和 4 0 μmol·L-1Cur均可使 2 5～ 4 0 0 μmol·L-1H2 O2 作用 2 4h后对PC12细胞生长的抑制率下降 ,可明显抑制 10 0和 2 0 0 μmo·L-1H2 O2 作用 2 4h后对PC12细胞凋亡的诱导作用和对PC12细胞△Ψm的降低作用 ,可明显降低 10 0和 2 0 0 μmol·L-1H2 O2 作用 12h后细胞内ROS的含量。结论 Cur对氧化应激损伤PC12细胞具有保护作用 ,其机制可能与降低细胞内ROS的含量 ,进而抑制△Ψm的降低有关。

AIM To study the protective effect of curcumin (Cur) on PC12 cells damage induced by oxidative stress. METHODS Using H 2O 2-induced PC12 cells damage as the model of neuron damage induced by oxidative stress, the proliferation of PC12 cells was observed by 3-[4,5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide(MTT)assay, the apoptosis of PC12 cells was detected by propidium iodide stain flow cytometry (FCM), the mitochondrial membrane potential (△Ψm) was analyzed by rhodamine123 stain FCM and the level of reactive oxygen species (ROS) in PC12 cells was analyzed by dihydrohodamine123 stain FCM. RESULTS In the presence of Cur (20 and 40 μmol·L -1), the inhibitory rates of PC12 cells induced by H 2O 2 from 25 to 400 μmol·L -1 and the apoptosis of PC12 cell induced by 100 and 200 μmol·L -1 H 2O 2 for 24 h were decreased. The level of △Ψm was decreased significantly in PC12 cells after 100 and 200 μmol·L -1 H 2O 2 treatment for 24 h, however, those decreases were significantly ameliorated by Cur (20 and 40 μmol·L -1) treatment. The level of ROS was significantly increased in PC12 cells exposed to H 2O 2 (100 and 200 μmol·L -1) for 12 h, whereas 40 μmol·L -1 of Cur prevented the rise induced by H 2O 2. CONCLUSIONS Cur has protective effect on PC12 cells damage induced by oxidative stress and the effect might be attributed to its removal of ROS and increase of △Ψm level.