Objective To observe the effects of Gandouling on reactive oxygen species (ROS) level, the expression of nuclear factor erythroid 2-related factor 2 (Nrf2) mRNA and protein of neural stem cells of the mice cultured in high concentration copper. Methods The model of neural stem cells of the mice was cultured in vitro with high concentration copper. The experimental rats were randomly divided into blank control group, model group, and Gandouling low-, medium-, and high-dose groups. Each medication group was given relevant concentration of Gandouling serum for gavage. The MTT was adopted to test proliferation level on neural stem cells; flow cytometer was used to examine the change of ROS level in cells; qPCR was used to measure the expression of Nrf2 mRNA;Western blot was used to measure the change of the level of protein Nrf2 in cells. Results Compared with the blank control group, the proliferation rate of neural stem cells was significantly decreased, ROS levels were significantly increased, and Nrf2 gene and protein expression was significantly decreased (P＜0.01). Compared with the model group, neural stem cells proliferation rate was significantly increased, ROS levels were significantly reduced, and Nrf2 gene and protein expression was significantly increased (P＜0.05, P＜0.01). Conclusion Gandouling can promote the proliferation of neural stem cells in mice by reducing ROS content in high copper-loaded mice and up-regulating Nrf2 expression.%目的 观察肝豆灵对高铜负荷小鼠神经干细胞内活性氧(ROS)水平及核因子E2相关因子2(Nrf2) mRNA 及蛋白表达的影响.方法 体外培养高铜负荷小鼠神经干细胞模型.实验小鼠随机分为空白对照组、模型组和肝豆灵低、中、高剂量组,各给药组给予相应浓度肝豆灵药物血清灌胃.采用MTT法检测神经干细胞增殖水平,流氏细胞仪检测细胞内ROS水平,qRT-PCR检测Nrf2基金表达,Western blot检测细胞内Nrf2蛋白表达.结果 与空白对照组比较,模型组小鼠神经干细胞增殖率显著下降,ROS水平明显升高,Nrf2基因、蛋白表达明显降低(P＜0.01);与模型组比较,肝豆灵各剂量组小鼠神经干细胞增殖率显著升高,ROS水平明显降低,Nrf2基因、蛋白表达明显升高(P＜0.05,P＜0.01).结论 肝豆灵通过降低高铜负荷小鼠ROS含量、上调Nrf2的表达,从而促进小鼠神经干细胞的增殖.
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