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[Baicalin increases the antioxidant capacity via promoting the nuclear translocation of NF-E2-related factor 2 (Nrf2) in N2a/APPswe cells].

黄芩苷 化学 活力测定 MTT法 分子生物学 丙二醛 超氧化物歧化酶 污渍 抗氧化剂 生物化学 细胞 生物 色谱法 高效液相色谱法 基因
作者
Huimin Cao,Beibei Chen,Yushuang Deng,Xi Lu,Gang Yu
出处
期刊:PubMed 卷期号:31 (12): 1597-601 被引量:6
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摘要

To investigate the protective effect and related mechanism of baicalin in murine neuroblastoma N2a cells stably expressing human Swedish mutant amyloid precursor protein (APP) (N2a/APPswe cells), a cellular model of Alzheimer' s disease (AD).MTT assay was performed to observe the effect of baicalin (0.1, 0.5, 1, 5, 10, 20) μmol/L on the viability of N2a/APPswe cells. After N2a/APPswe cells were incubated with (1, 5, 10) μmol/L baicalin for 48 hours, xanthine oxidase assay was used to test superoxide dismutase (SOD) activity and thiobarbituric acid method to detect malondialdehyde (MDA) content in each group. Real-time quantitative PCR was applied to determine nuclear factor erythroid 2-related factor 2 (Nrf2) mRNA, and Western blotting to examine protein levels of total Nrf2, nuclear Nrf2 and nuclear factor κB (NF-κB) in N2a/APPswe cells exposed to different doses of baicalin. Immunofluorescence staining was also used to observe the distribution of Nrf2.We found that baicalin pretreatment increased cell viability. Compared with the control group (N2a/wt cells), SOD activity in N2a/APPswe cells significantly decreased, and MDA content significantly increased; but SOD activity was improved and MDA production was inhibited after pretreatment with baicalin, especially with 10 μmol/L bacalin. Both mRNA and total protein expression of Nrf2 were not significantly changed in baicalin treatment group compared with N2a/APPswe group, but the nuclear protein of Nrf2 distinctly increased after treatment with baicalin. In addition, baicalin decreased the level of nuclear NF-κB protein. Furthermore, immunofluorescence staining revealed that baicalin promoted the translocation of Nrf2 to the nucleus.Baicalin has the protection against oxidative stress via activation of Nrf2 in N2a/APPswe cells.

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