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《China Journal of Traditional Chinese Medicine and Pharmacy》 2016-05
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Astragaloside Ι inhibited the activation of BV-2 cell induced by LPS through modulation of PI3K/Akt/NF-κB pathway

LIU Hong-shuai;LI Wen-hui;SHEN Hui;TANG Yun-di;SHI Hai-lian;WU Hui;HUANG Fei;WU Xiao-jun;HU Zhi-bi;Shanghai Key Laboratory of Complex Prescription,Institute of Chinese Materia Medica,Shanghai University of Traditional Chinese Medicine;  
Objective: To investigate the effect and mechanism of astragaloside I(AST I) on the activation of microglial cell line BV-2 induced by LPS. Methods: After pre-treated with AST I(25, 50, 100μmol/L) for 2 h, BV-2 cells were stimulated with LPS(200ng/m L) for 20 h. Thereafter, the cells and the culture medium were collected. Cell viability was measured by CCK-8 method. Concentration of TNF-α was detected by using ELISA kit. Secretion of NO in medium was assayed with Greiss approach. Gene expressions of CD11 b, TNF-α, i NOS, and IL-1β m RNA were detected with quantitative PCR. Protein expression of PI3K/Akt/NF-κB pathway was detected using Western blotting assay. Neuclear translocation of p-NF-κB was monitored with immunocytochemistry. Results: AST I inhibited the secretion of TNF-α and NO on BV-2 cells upon LPS stimulation(P0.001) without affecting cell viability. It could prevent the up-regulation of gene expressions of TNF-α, i NOS and IL-1β(P0.001, P0.001, P0.05) but not that of CD11 b. Meanwhile, AST I suppressed the increase of i NOS and COX-2 protein induced by LPS. Further study disclosed that AST I reduced nuclear translocation of activated NF-κB and deceased phosphorylation of PI3 K, Akt, NF-κB, and IκB. Conclusion: AST I inhibited the activation of BV-2 cells induced by LPS through suppressing the activation of PI3K/Akt/NF-κB pathway, therefore, reduced the nuclear translocation of phosphorylated NF-κB, leading to the down-regulation of the gene expressions of TNF-α, i NOS and IL-1β and thus lessened the production of i NOS, TNF-α and COX-2 protein.
【Fund】: 上海高校特聘教授(东方学者)岗位计划(No.2013-59);; 上海高校中药药效物质E研究院项目~~
【CateGory Index】: R285
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