山柰酚对脂多糖诱导小胶质细胞炎性介质释放与吞噬的影响

Inhibitory effect of kaempferol on lipopolysaccharide-induced inflammatory mediator release and phagocytosis of microglia

目的:研究山柰酚(KF)对脂多糖(LPS)诱导的BV2小胶质细胞炎性介质释放及吞噬能力的影响。方法:用LPS 10μg/ml激活BV2小胶质细胞,MTT法检测不同浓度山柰酚(0、10、20、40、80、160、320μmol/L)处理后BV2小胶质细胞的细胞活性,确定山柰酚给药浓度。BV2细胞分为5组:正常组(NS)、脂多糖激活BV2小胶质细胞组(LPS)、山柰酚10μmol/L低浓度处理组(LPS+KF-L)、山柰酚20μmoL/L中浓度处理组(LPS+KF-M)和山柰酚40μmol/L高浓度处理组(LPS+KF-H)。real time RT-PC R检测炎症因子白介素6(IL-6)、白介素8(IL-8)、白介素1β(IL-1β)和肿瘤坏死因子α(TNF-α)的mRNA水平;荧光乳胶珠吞噬实验测定活化的BV2小胶质细胞经不同浓度山柰酚(10、20、40μmol/L)作用后的吞噬功能。结果:MTT显示山柰酚在10~40μmol/L浓度范围内对BV2细胞没有毒性作用、不同浓度山柰酚能够剂量依赖性抑制LPS诱导的BV2细胞中IL-6、IL-8、IL-1β和TNF-αmRNA的表达,抑制小胶质细胞的吞噬功能。结论:山柰酚抑制炎性介质释放,抑制小胶质细胞吞噬。

Objective: To study the effect of kaempferol( KF) on the release of inflammatory mediators and phagocytosis of BV2 cells induced by lipopolysaccharide( LPS). Methods: BV2 cells were activated with LPS 10 μg/ml.The MTT assay was used to detect the cell viability of different concentrations of kaempferol( 0,10,20,40,80,160,320 μmol/L) on BV2 cells,then determine the optimum concentration of kaempferol. BV2 cells were divided into five groups: BV2 cells normal group( NS),lipopolysaccharide-activated BV2 cells group( LPS),10 μmol/L kaempferol low concentration group( LPS + KF-L),20 μmol/L kaempferol medium concentration group( LPS + KF-M),40 μmol/L kaempferol concentration group( LPS + KF-H). The mRNA levels of the inflammatory factors interleukin 6( IL-6),interleukin8( IL-8),interleukin1( IL-1) and tumor necrosis factor-α( TNF-α) were detected by real-time quantitative fluorescence PCR. The phagocytic function of activated BV2 cells was tested by fluorescent latex bead phagocytosis assay after treated with different concentrations of kaempferol( 10,20,40 μmol/L). Results: MTT test showed that kaempferol had no toxic effect on BV2 cells in the concentration range of 10 ~ 40 μmol/L. kaempferol could dose-dependently significantly inhibited IL-6,IL-8,IL-1β,and TNF-α mRNA expression by LPS-induced BV2 cells and inhibited phagocytosis. Conclusion: Kaempferol inhibits the release of inflammatory mediators and inhibits the phagocytosis of BV2 cells.