摘要
目的探讨阿曼托双黄酮(AF)对脂多糖(LPS)诱导的BV-2小胶质细胞的炎性因子的阻断效应。方法首先通过CCK-8法筛选出对BV-2细胞活性无影响的AF浓度;在此基础上,采用10mol/L AF预处理BV-2小胶质细胞1 h,加入1.0g/mL LPS诱导细胞炎症反应,通过实时荧光定量PCR检测炎性因子白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)、诱导型一氧化氮合酶(iNOS)及环加氧酶2(COX2)的mRNA水平,Western blot法检测iNOS及COX2的蛋白水平,免疫荧光细胞化学染色法检测COX2、 iNOS的表达和定位。结果 CCK-8法证明10μmol/L AF对BV-2小胶质细胞活性无明显影响。LPS单独处理可增加BV-2小胶质细胞IL-1β、 TNF-α、 COX2、 iNOS的mRNA表达及COX2、 iNOS蛋白表达;与LPS组相比,10μmol/L AF预给药组能够明显降低IL-1β、 TNF-α、 COX2、 iNOS的mRNA水平及COX2和iNOS的蛋白水平,同时AF可明显抑制COX2及iNOS的表达,抑制BV-2小胶质细胞的激活状态。结论 AF对LPS诱导的BV-2小胶质细胞炎症反应具有保护作用。
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.
作者
任晓璠
张庆鹏
荣世阔
左娣
王峰
刘昆梅
REN Xiaofan;ZHANG Qingpeng;RONG Shikuo;ZUO Di;WANG Feng;LIU Kunmei(Ningxia Key Laboratory of Cerebrocranial Diseases,Ningxia Medical University,Yinchuan 750004;Shizuishan Municipal First Peopled Hospital,Shizuishan 750002,China)
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2020年第1期14-19,共6页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金(81460108)
宁夏自治区“十三五”重大科技项目(2016BZ07)
宁夏医科大学校级科研项目(XY2017020)。
