Nrf2通过上调自噬促进C17.2神经干细胞的增殖（英文）  

Nrf2 Promotes C17.2 Neural Stem Cell Proliferation via Up-Regulating Autophagy

作　　者：杨晓红[1] 王胜男[1] 刘庆余[2] 欧阳颖[2] 李民[1] 皮荣标[1] 

机构地区：[1]中山大学药学院,广东广州510006 [2]中山大学孙逸仙纪念医院,广东广州510120

出　　处：《中山大学学报（医学科学版）》2016年第4期481-489,共9页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(31371070);广东省国际科技合作项目(2013B051000038);中央高校基础研究项目(15ykjc08b);广东省自然科学基金(S2013010016308)

摘　　要：【目的】通过使用化合物调控细胞内信号通路,最终引起脑内神经干细胞的增殖和分化。这被为认为是一种崭新的修复受损大脑功能的治疗策略。我们发现Nrf2能够调控C17.2神经干细胞的增殖,并试图阐明其可能的作用机制。【方法】采用MTT法检测细胞活力,Brd U掺入法检测细胞增殖,western blot法检测细胞内蛋白水平的变化,流式细胞术检测细胞内ROS水平的变化。【结果】Nrf2的激动剂t-BHQ和SFN可以促进C17.2神经干细胞的增殖,而使用Nrf2的抑制剂brusatol则抑制其增殖。然而无论是Nrf2的激动剂还是抑制剂均对细胞内的ROS水平没有影响,但是通过t-BHQ和SFN激活Nrf2能降低H2O2导致的C17.2细胞损伤。此外,我们观察到t-BHQ和SFN能增加细胞内LC3Ⅱ的蛋白水平以及GFP-LC3点子的数目,而brusatol则具有相反的作用。过表达Nrf2不仅导致LC3Ⅰ向Ⅱ的转变,还能促进C17.2细胞的增殖,而加入自噬的抑制剂NH4Cl和CQ则能部分逆转Nrf2导致的C17.2细胞增殖。【结论】Nrf2能够促进C17.2干细胞的增殖,这可能与其升高细胞内的自噬水平相关,而不是与细胞内ROS的水平相关。[ Objective ] In order to repair or restore the function of damaged brain, to regulate signaling pathway by chemicals, which ultimately leads to endogenous neural stem cells （NSC） proliferation and differentiation, has been regarded as an new treatment strategy. Here, we found the effects of nuclear factor erythroid 2-related factor （Nrf2） on C17.2 cell proliferation, and tried to illuminate its possible mechanisms. [ Methods ] Cell viability was tested by Mrl＇r assay, and cell proliferation was measured by BrdU incorporation assay. Intracellular ROS formation was monitored by flow eytometry. The protein levels were measured by westem blot. [ Results ] Two Nrf2 activators, tert-butylhydroquinone （t-BHQ） and sulforaphane （SFN）, promoted the proliferation of C 17.2 neural stem cells, which was suppressed by brusatol, an Nrf2 inhibitor. Unexpectedly, intraeellular reactive oxygen species （ROS） after treatment with Nrf2 activators or inhibitor were similar to the control group, but Nrf2 activation by t-BHQ or SFN could protect C17.2 cells against hydrogen peroxide （H202）-induced damage. Moreover, we observed that microtubule-assoeiated protein 1 light chain 3 （LC3）-II protein level and the green fluorescent protein tagged-LC3 （GFP-LC3） puneta were markedly increased by treatment with t- BHQ or SFN and were significantly decreased by exposure to brusatol in C17.2 ceils. Overexpression of Nrf2 not only induced the conversion of LC3-I to LC3-II, but increased C17.2 cell proliferation that was partly reversed by two autophagy inhibitors,, ammonium chloride （NH4C1） and ehloroquine （CQ）. [ Conclusion ] Our findings suggest that activating autophagy, instead of modulating ROS, plays a pivotal role in the Nrf2-induced C17.2 cell proliferation.

关 键 词：NRF2 神经干细胞 增殖 ROS 自噬 

分 类 号：R966[医药卫生—药理学]