利福平预处理通过Nrf2减轻帕金森病小胶质细胞的氧化损伤  

Rifampicin Pretreatment Inhibits Oxidative Damage of Microglia via Nrf2 Signaling Pathway in Parkinson's Disease

作　　者：梁嫣然[1] 林淡钰[1] 陈颖[1] 毕伟[2] 曾志芬[1] 井秀娜[1] 伍霞[1] 陶恩祥[1] 

机构地区：[1]中山大学孙逸仙纪念医院神经内科,广东广州510120 [2]暨南大学附属第一医院神经内科,广东广州510630

出　　处：《中山大学学报（医学科学版）》2016年第2期190-196,共7页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(81503052);广东省医学科研基金(B2014130);国家级大学生创新创业训练计划项目(201410558098);广东省自然科学基金(2014A030313202);2014年广东省公益研究与能力建设专项资金(2014A020212164)

摘　　要：【目的】探讨利福平对鱼藤酮诱导的BV2小胶质细胞氧化损伤中的核因子E2相关因子2(Nrf2)信号通路的影响。【方法】利用鱼藤酮刺激的BV2小胶质细胞建立帕金森病氧化应激的模型,分别用不同浓度的利福平预处理BV2小胶质细胞,CCK-8法检测细胞存活率;分别用双氯荧光素(DCFH-DA)、罗丹明123染色后使用流式细胞仪检测活性氧(ROS)水平和线粒体膜电位(MMP);使用脂质氧化检测试剂盒测定细胞内脂质过氧化终产物丙二醛(MDA)的含量。采用Western Blot法检测利福平对小胶质细胞Nrf2核转位以及血红素加氧酶-1(HO-1)蛋白表达的影响。用si RNA抑制HO-1基因表达后用CCK-8法检测利福平对鱼藤酮诱导的氧化损伤的影响。【结果】与空白对照组相比,鱼藤酮刺激下BV2小胶质细胞的细胞活力明显降低。鱼藤酮可引起BV2细胞内ROS和MDA的水平显著升高以及MMP明显下降。利福平预处理可以降低鱼藤酮所引起的ROS和MDA的产生,以及逆转鱼藤酮对MMP的降低作用;利福平能够诱导Nrf2的核转位及上调HO-1蛋白的表达。使用si RNA抑制HO-1基因表达后,可逆转利福平预处理对鱼藤酮诱导的氧化损伤的抑制作用。【结论】利福平可能通过激活Nrf2信号通路,减轻鱼藤酮诱导的帕金森病小胶质细胞模型的氧化损伤。[ Objective ] To investigate the effect of rifampicin on Nff2 signal pathway during protection against rotenone-induced oxidative damage in BV2 microglia. [Methods] The in vitro model to assess the mechanism of oxidative stress in Parkinson＇s disease （PD） was established by adding rotenone to BV2 cells.The cell viability was measured by CCK-8 assay. Furthermore,the intracellular reactive oxygen species （ROS）and mitochondrial membranepotential （MMP） were detected by flow cytometry after dyed with DCFH- DA and Rhodamine123, respectively. The level of malondialdehyde （MDA） was measured according to the protocol of Lipid Oxidation Products Detection Kit. The effects of rifampicin on nuclear translocation of Nrf2 and protein expression of hemeoxygenase-1 （HO-1 ） were measured by western blot. After transfection with siRNA targeting HO-1 gene, the effect of rifampiein on rotenone-induced eytotoxicity was evaluated by CCK-8 assay. [Resuhs] Compared with the control group, the cell viability of BV2 cells decreased obviously, and intraeellular ROS and MDA levels increased significantly, but MMP decreased markedly in rotenone group. However, such trends could be reversed in the rifampicin pretreatment group. Moreover, rifampicin could induce nuclear translocation of Nrf2 and significantly increase protein expression of HO-1 when compared with rotenone group. After knocking down the expression of HO- 1 using siRNA, the inhibitory effect of rifampicin on cytotoxicity induced by rotenone was reversed. [ Conclusion ] Rifampicin may protect against damage of oxidative stress in PD via activation of Nrf2 signaling pathway.

关 键 词：帕金森病 利福平 鱼藤酮 核因子E2相关因子2 小胶质细胞 

分 类 号：R74[医药卫生—神经病学与精神病学]