人参皂甙Rd减轻H2O2介导的小胶质细胞氧化应激损伤

Ginsenoside Rd reduces oxidative stress damage induced by H2O2 in microglia cells

目的探讨人参皂甙Rd能否对抗H2O2刺激小胶质细胞引起的氧化应激损伤及对小胶质细胞生物功能的影响。方法实验分为空白对照组、人参皂甙Rd组、H202组、人参皂甙Rd10ixmol／L＋H202组、人参皂甙Rd20ixmol／L＋H202组。空白对照组为小鼠小胶质细胞系N9细胞在CO2孵箱孵育24h；人参皂甙Rd组应用20μmol／L人参皂甙Rd处理N9细胞24h；H202组应用700Ixmol／LH202处理N9细胞24h；人参皂甙Rd10ixmol／L＋H202组、人参皂甙Rd20ixmol／L＋H202组分别应用10txmol／L、20txmol／L人参皂甙Rd预处理N9细胞24h，然后加入700iμmol／LI-1202作用24h。处理结束后，应用MTT法检测各组细胞活力，应用2＋．7＋．二氯荧光素二乙酸酯（DCFH-DA）检测细胞内活性氧（ROS）水平，应用四甲基罗丹明乙酯（TMRE）检测线粒体膜电位（MMP），应用Westemblotting检测脑源性神经营养因子（BDNF）蛋白表达水平。结果与空白对照组、人参皂甙Rd组相比，H2O2组细胞存活率明显下降，ROS水平明显升高，MMP水平明显降低，差异均有统计学意义（氏0．05）；与空白对照组相比，人参皂甙Rd组BDNF蛋白表达水平升高，差异有统计学意义（P〈0．05）；与H202组相比，人参皂甙Rd10μmol／L＋H20：组、人参皂甙Rd20μmol／L＋H2O2组细胞存活率明显升高，ROS水平明显下降，MMP水平明显增高，BDNF蛋白表达水平增高，差异均有统计学意义（P〈0．05）。结论人参皂甙Rd能够对抗H：0：介导的小胶质细胞氧化应激损伤，且能促进小胶质细胞分泌BDNF，对小胶质细胞有显著的保护作用。

Objective To investigate the effect ofginsenoside Rd on oxidative stress induced by H202 in microglia cells and the changes of biological function of microglia cells. Methods The microglia cell lines N9 were divided into control group, ginsenoside Rd group, H202 group, ginsenoside Rd 10 μmol/L＋H202 group and ginsenoside Rd 20 I-mol/L＋H202 group. In the control group, the N9 cells were incubated in the CO2 incubator for 24 h; in the ginsenoside Rd group, the N9 cells were treated with 20 μmol/L ginsenoside Rd for 24 h; in the H202 group, the N9 cells were treated with 700 -mol/L H202 for 24 h; the cells in ginsenoside Rd 10 txmol/L＋H202 group and ginsenoside Rd 20 -mol/L＋ H202 group were pretreated with 10 p.mol/L or 20 p-mol/L ginsenoside Rd for 24 h, respectively, and then, 700 -mol/L H202 was added to the medium and co-cultured for another 24 h. After the treatment, the cell viability was measured by MTT assay, the intracellular reactive oxygen species （ROS） level was determined by 2＇,7＇-dichlorofluorescein diacetate （DCFH-DA） and mitochondrial membrane potential （MMP） was measured by tetramethylrhodamine methyl ester （TMRE）. Western blotting was used todetect the expression of brain derived neurotrophic factor （BDNF） in N9 cells. Results As compared with those in the control group and ginsenoside Rd group, the cell survival rate was significantly decreased, ROS level was significantly increased and MMP was significantly decreased in H：O2 group （P〈0.05）; as compared with the control group, the BDNF expression in ginsenoside Rd group was higher, and the difference was statistically significant （P〈0.05）; as compared with H202 group, the cell survival rate, BDNF expression and MMP level increased significantly, ROS was significantly decreased in ginsenoside Rd 10 Ixmol/L＋H202 group and ginsenoside Rd 20 pomol/L＋H2Oz group （P〈0.05）. Conclusion Ginsenoside Rd could protect microglia cells from oxidative stress damage induced by H202, and could increase the secretion of BDNF,which has a significant protective effect on microglia cells.