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目的探讨黄芩苷对阿尔茨海默病(AD)转基因N2a/APPswe细胞的保护作用及可能机制。方法应用(0.1、0.5、1、5、10、20)μmol/L黄芩苷处理N2a/APPswe细胞,MTT法检测黄芩苷对细胞存活率的影响。用(1、5、10)μmol/L黄芩苷处理细胞48 h,通过黄嘌呤氧化酶法检测各组细胞超氧化物歧化酶(SOD)的活性,硫代巴比妥酸法检测各组细胞培养上清液的丙二醛(MDA)的含量。荧光实时定量PCR测定核因子E2相关因子2(Nrf2)的mRNA表达,Western blot法检测细胞总Nrf2、核内Nrf2和核因子κB(NF-κB)的蛋白水平。免疫荧光染色观察Nrf2在细胞内的分布。结果 MTT试验结果显示,黄芩苷能促进N2a/APPswe细胞增殖。与对照组相比,模型组SOD活性明显降低,MDA含量明显增多,而黄芩苷能提高SOD活性,抑制MDA的生成,尤其以高剂量组最为明显。黄芩苷处理组与模型组相比Nrf2 mRNA和总蛋白表达均无明显差异,但黄芩苷处理后能显著增加细胞核内Nrf2的含量,下调细胞核内NF-κB的水平。免疫荧光染色结果也证明,黄芩苷能促进Nrf2的核转位。结论黄芩苷增强N2a/APPswe细胞的抗氧化能力并促进Nrf2的核转位。
Objective To investigate the protective effect and possible mechanism of baicalin on Alzheimer’s disease (AD) transgenic N2a / APPswe cells. Methods N2a / APPswe cells were treated with (0.1,0.5,1,5,10,20) μmol / L baicalin, and the effects of baicalin on cell viability were detected by MTT assay. The cells were treated with (1,5,10) μmol / L baicalin for 48 h. The activity of superoxide dismutase (SOD) in each group was detected by xanthine oxidase method. The cells in each group were detected by thiobarbituric acid method The supernatant was cultured for malondialdehyde (MDA) content. Real-time quantitative PCR was used to detect the mRNA expression of nuclear factor E2-related factor 2 (Nrf2). Western blot was used to detect the protein levels of Nrf2, Nrf2 and NF-κB. Immunofluorescence staining to observe the distribution of Nrf2 in the cell. Results MTT assay showed that baicalin can promote the proliferation of N2a / APPswe cells. Compared with the control group, the activity of SOD in the model group decreased significantly, and the MDA content increased obviously. Baicalin increased the activity of SOD and inhibited the formation of MDA, especially in the high dose group. There was no significant difference in the expression of Nrf2 mRNA and total protein between the baicalin treatment group and the untreated group. However, baicalin treatment could significantly increase the content of Nrf2 in the nucleus and decrease the level of NF-κB in the nucleus. Immunofluorescence staining results also proved that baicalin can promote Nrf2 nuclear translocation. Conclusion Baicalin can enhance the antioxidant capacity of N2a / APPswe cells and promote the nuclear translocation of Nrf2.