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探讨没药甾酮(guggulsterone)对氧化应激损伤PC12细胞的保护作用。以过氧化氢(hydrogen peroxide,H2O2)损伤PC12细胞为氧化应激损伤模型,维生素E为对照,采用四甲基偶氮唑蓝[3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl-tetrazolium bromide,MTT]法检测细胞增殖状况;试剂盒检测乳酸脱氢酶(lactate dehydrogenase,LDH)及一氧化氮(nitric oxide,NO)的释放;DCFH法和Fura 2-AM法检测细胞内活性氧(reactive oxygen species,ROS)和Ca2+的含量;碘化丙啶(propidium iodide,PI)染色流式细胞术(flow cytometry,FCM)检测细胞凋亡;罗丹明123(rhodamine 123,Rh 123)染色FCM检测细胞线粒体膜电位(mitochondrial membrane protential,MMP)。结果表明,没药甾酮(0.1～10μmol.L-1)可使200μmol.L-1H2O2作用24 h后的PC12细胞生长抑制率下降;细胞外LDH和NO,细胞内ROS和Ca2+含量降低;明显抑制200μmol.L-1H2O2作用12 h后诱导的PC12细胞凋亡和线粒体膜电位降低作用,没药甾酮(0.1～10μmol.L-1)使细胞凋亡率由24.3%下降至18.4%、15.9%、11.8%。实验结果表明,没药甾酮对氧化应激损伤PC12细胞具有保护作用,其机制可能为降低细胞内ROS含量,进而抑制LDH和NO释放,降低细胞内Ca2+含量,升高线粒体膜电位,减少细胞凋亡。
To investigate the protective effect of guggulsterone on PC12 cells injured by oxidative stress. Hydrogen peroxide (H2O2) was used to damage PC12 cells as an oxidative stress injury model. Vitamin E was used as a control and tetramethylazolyl blue [3-(4,5-dimethylthiazole-2-yl)-2] was used. 5-diphenyl-tetrazolium bromide (MTT) method was used to detect cell proliferation. The kit was used to detect the release of lactate dehydrogenase (LDH) and nitric oxide (NO). DCFH assay and Fura 2-AM assay The content of reactive oxygen species (ROS) and Ca2+ in cells; propidium iodide (PI) staining flow cytometry (FCM) to detect apoptosis; rhodamine 123 (rhodamine 123, Rh 123) Staining FCM to detect mitochondrial membrane potential (MMP). The results showed that the mycelial dose (0.1～10μmol.L-1) could reduce the growth inhibition rate of PC12 cells treated with 200μmol.L-1H2O2 for 24 h; the extracellular LDH and NO, intracellular ROS and Ca2+ contents decreased; The apoptosis of PC12 cells and the decrease of mitochondrial membrane potential induced by the inhibition of 200μmol.L-1H2O2 for 12 h, the apoptotic rate of mycelia (0.1-10 μmol.L-1) decreased from 24.3% to 18.4% and 15.9. %, 11.8%. The experimental results show that myrrhoside has a protective effect on PC12 cells injured by oxidative stress. The mechanism may be to decrease intracellular ROS content, thereby inhibit the release of LDH and NO, decrease the intracellular Ca2+ content, increase the mitochondrial membrane potential, and reduce the number of cells. Apoptosis.