枫叶黄酮抑制脂多糖诱导破骨前体细胞Raw264.7细胞激活的作用

Flavonoid Isolated from the Leaves of Acer Truncatum Bunge Inhibits Lipopolysaccharide-induced Activation of Raw264.7 Cells

目的探讨枫叶黄酮对脂多糖(lipopolysaccharide,LPS)诱导破骨前体细胞Raw264.7细胞炎性因子释放的抑制作用.方法用LPS(5 mg/mL)刺激Raw264.7细胞构建炎症模型,采用四甲基偶氮唑蓝比色法(MTT)检测Raw264.7细胞的活力影响;酶联免疫吸附试验(ELISA)检测一氧化氮(NO)和前列腺素E2,PGE2表达;运用免疫荧光染色方法检测诱导型一氧化氮合酶和环氧合酶-2,COX-2、子肿瘤坏死因子-αTNF-α和白介素-1β,IL-1β、核因子-κB,NF-κB蛋白与mRNA的表达变化.结果不同浓度的枫叶黄酮明显抑制了LPS诱导的Row246.7细胞NO、PGE2,iNOS和COX-2,TNF-α和IL-1β与NF-κB的上调,并且NO、PGE2、iNOS、COX-2和NF-κB蛋白的表达呈剂量依赖性.结论枫叶黄酮可抑制LPS诱导的破骨前体细胞Raw264.7细胞iNOS、COX-2和NF-κB/P65蛋白表达和炎性因子释放从而抑制破骨细胞的激活.

Objective To investigate the inhibitory effects of flavonoid isolated from the leaves of Acer truncatum Bunge on the expression of pro-inflammatory cytokines in Raw264.7 cells stimulated with lipopolysaccharide（LPS）.Methods Inflammatory cell model was structured by LPS-stimulated Raw264.7 cells,the cells were treated with flavonoid（10 μM,30 μM and 60 μM） prior to LPS（5 mg/mL）exposure and the effects of flavonoid on viability of Raw264.7 cells were measured by MTT assay,Nitric oxide（NO）and prostaglandin E2（PGE2）were measured by ELISA,the effects on the protein levels of pro-inflammatory enzymes,cyclooxygenase-2（COX-2）and inducible nitric oxide synthase（iNOS）,and pro-inflammatory cytokines,tumor necrosis factor-α（TNF-α）,and interleukin-1β（IL-1β）,inflammatory signaling proteins nuclear factor-κB（NF-κB）were analysed by immunofl uorescence staining assay.Results LPS-induced NO,PGE2,iNOS,COX-2 and NF-κB protein expression levels were significantly decreased by flavonoid in a concentration-dependent manner and flavonoid also had an effect on the expression of TNF-α,IL-1β through translational inhibition.Conclusion Flavonoid isolated from the leaves of Acer truncatum Bunge can inhibit LPS-induced activation of Raw264.7 cells through inhibiting the release of pro-infl ammatory mediators,then exerts its anti-osteoporosis activities.