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[穗花杉双黄酮抑制脂多糖诱导的小鼠BV-2小胶质细胞炎症反应]

[Amentoflavone inhibits inflammation of mouse BV-2 microglia cells induced by lipopolysaccharide].

作者信息

Ren Xiaofan, Zhang Qingpeng, Rong Shikuo, Zuo Di, Wang Feng, Liu Kunmei

机构信息

Ningxia Key Laboratory of Cerebrocranial Diseases, Ningxia Medical University, Yinchuan 750004, China.

Shizuishan Municipal First People's Hospital, Shizuishan 750002, China.

出版信息

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2020 Jan;36(1):14-19.

Abstract

Objective To investigate the block effect of amentoflavone (AF) on the inflammation of mouse BV-2 microglial cells induced by lipopolysaccharide (LPS). Methods BV-2 microglial cells were treated with AF at different concentrations, and cell viability was determined by CCK-8 assay to get the AF concentration that had no effect on the cell viability. BV-2 microglia cells were pretreated with 10 mol/L AF, and 1 hour later, 1.0 g/mL LPS was used to induce inflammatory response in the BV-2 microglial cells. Real-time quantitative PCR was performed to detect the gene expression of interleukin 1β (IL-1β), tumor necrosis factor α (TNF-α), cyclooxygenase 2 (COX2) and inducible nitric oxide synthase (iNOS). The protein expression of COX2 and iNOS were measured by Western blot analysis. Immunofluorescence staining was used to observe the location and expression of COX2 and iNOS. Results CCK-8 showed that 10 mol/L AF did not affect the viability in BV-2 microglial cells. The treatment of 1.0 g/mL LPS could significantly up-regulate the mRNA expression of IL-1β, TNF-α, COX2, iNOS, and the protein expression of COX2 and iNOS. Compared with the only LPS treatment, 10 mol/L AF pretreatment markedly decreased the elevated gene and protein expression induced by LPS. In addition, AF significantly inhibited the expression of COX2 and iNOS, and less microglial cells were activated. Conclusion AF can inhibit the inflammation of BV-2 microglial cells induced by LPS.

摘要

目的 探讨穗花杉双黄酮(AF)对脂多糖(LPS)诱导的小鼠BV-2小胶质细胞炎症的阻断作用。方法 用不同浓度的AF处理BV-2小胶质细胞,通过CCK-8法检测细胞活力,以获得对细胞活力无影响的AF浓度。用10 μmol/L AF预处理BV-2小胶质细胞,1小时后,用1.0 μg/mL LPS诱导BV-2小胶质细胞发生炎症反应。采用实时定量PCR检测白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)、环氧化酶2(COX2)和诱导型一氧化氮合酶(iNOS)的基因表达。通过蛋白质印迹分析检测COX2和iNOS的蛋白表达。采用免疫荧光染色观察COX2和iNOS的定位及表达。结果 CCK-8结果显示,10 μmol/L AF不影响BV-2小胶质细胞的活力。1.0 μg/mL LPS处理可显著上调IL-1β、TNF-α、COX2、iNOS的mRNA表达以及COX2和iNOS的蛋白表达。与仅用LPS处理相比,10 μmol/L AF预处理显著降低了LPS诱导的基因和蛋白表达升高。此外,AF显著抑制COX2和iNOS的表达,且活化的小胶质细胞较少。结论 AF可抑制LPS诱导的BV-2小胶质细胞炎症。

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