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目的:研究金雀异黄素对脂多糖(LPS)诱导巨噬细胞RAW264.7分泌功能的影响,探讨金雀异黄素抗炎作用及其机制,为临床应用提供理论基础。方法:用脂多糖诱导巨噬细胞株Raw264.7建立体外细胞炎症模型,金雀异黄素与巨噬细胞株Raw264.7温孵1h后,加入100ng·mL-1脂多糖以诱导巨噬细胞的活化,采用Griess试剂检测细胞培养液中NO的含量,通过双抗体夹心酶联分析法和细胞转染技术观察不同浓度金雀异黄素对脂多糖刺激巨噬细胞分泌细胞因子TNF-α、IL-6和IL-10含量及对核转录因子NF-κB和AP-1表达的影响。结果:金雀异黄素(4～111μmolL-1)能明显抑制脂多糖刺激巨噬细胞生成一氧化氮产生(P<0.05);显著抑制脂多糖诱导巨噬细胞分泌细胞因子TNF-α、IL-6和IL-10(P<0.01),并能抑制核转录因子NF-κB和AP-1的表达。结论:本实验证明了金雀异黄素有抗炎作用,其能显著降低脂多糖诱导巨噬细胞分泌NO及细胞因子TNF-α、IL-6和IL-10的释放,通过抑制这些炎症因子的释放来达到抗炎作用,金雀异黄素抑制核转录因子NF-κB和AP-1的表达来实现的。
Objective: To investigate the effect of genistein on lipopolysaccharide (LPS) -induced macrophage RAW264.7 secretion, explore the anti-inflammatory effect of genistein and its mechanism, and provide a theoretical basis for clinical application. Methods: The model of cell inflammation was established by inducing macrophage cell line Raw264.7 with lipopolysaccharide. Genistein and macrophage cell line Raw264.7 were incubated for 1 hour, then 100 ng · mL-1 lipopolysaccharide was added to induce macrophage Griess reagent was used to detect the content of NO in cell culture medium. The effects of different concentrations of genistein on lipopolysaccharide-stimulated macrophage-secreted cytokine TNF-α were observed by double antibody sandwich enzyme-linked assay and cell transfection technique. IL-6 and IL-10 content and the expression of nuclear factor-κB and AP-1. Results: Genistein (4 ~ 111μmolL-1) significantly inhibited LPS-induced macrophage NO production (P <0.05), and significantly inhibited LPS-induced macrophage secretion of cytokines TNF-α, IL -6 and IL-10 (P <0.01), and inhibited the expression of NF-κB and AP-1. CONCLUSION: This experiment demonstrated that genistein has an anti-inflammatory effect which can significantly reduce the release of NO and cytokines TNF-α, IL-6 and IL-10 secreted by macrophages by lipopolysaccharide. By inhibiting these inflammatory factors Of the release to achieve anti-inflammatory effect, genistein inhibition of nuclear transcription factor NF-κB and AP-1 expression to achieve.