苯并噁唑类化合物K310对LPS刺激RAW264.7巨噬细胞的抗炎作用

目的 探索K310对脂多糖(LPS)诱导RAW264.7巨噬细胞的抗炎作用及机制.方法 在LPS诱导RAW264.7巨噬细胞的模型基础上,采用CCK-8方法检测不同浓度K310对RAW264.7巨噬细胞活性的影响;Griess试剂检测K310对一氧化氮(NO)的影响;酶联免疫吸附法(ELISA)检测K310对IL-6炎性因子的影响;荧光定量PCR方法检测K310对诱导型一氧化氮(iNOS)和IL-6基因表达的影响;荧光免疫印迹法(Western bolt)检测K310对磷酸化的NF-kB和ERK1/2蛋白的影响.结果 K310(5、10和20 μM)不影响LPS刺激RAW264.7巨噬细胞的活性;与LPS刺激组比较,K310不仅能显著抑制NO和IL-6的炎性因子及炎性基因;还能抑制磷酸化的NF-kB和ERK1/2蛋白的表达.结论 K310对LPS诱导RAW264.7巨噬细胞的炎症具有抗炎作用,其作用机制可能是K310通过抑制NF-kB和MAPK信号通路,从而抑制NO和IL-6炎性因子的产生.

The Anti-inflammatory Effect of the Benzoxazole Compound K310 on the LPS-stimulated RAW264.7 Macrophages

Objective To investigate the anti-inflammatory effect and mechanism of K310 on lipopolysaccharide(LPS)-induced RAW264.7 macrophages.Methods Cell Counting Kit-8(CCK-8)was used to detect determine the effect of different concentration of K 310 on the activity of RAW264.7 macrophages induced by LPS.The Griess reagent was used to detect the effect of K 310 on nitric oxide(NO).The ELISA method was used to the effect of K310 on the inflammatory factors of IL-6.Fluorescence quantitative PCR was used to detect the effect of K310 on the expression of inducible nitric oxide synthase(iNOS)and IL-6.Western blot was used to detect the effect of K310 on phosphorylated NF-kB and phosphorylated ERK1/2 protein.Results K310 of 5μM,10 μM and 20 μM showed no effect on the activity of LPS-induced RAW264.7 macrophages.Compared with LPS-stimulated group,K310 significantly inhibited the inflammatory factors and mRNA levels of NO and IL-6,and it also inhibited the expression of phosphorylated NF-kB and ERK1/2 protein.Conclusion K310 has the anti-inflammatory effect on LPS-induced RAW264.7 macrophages and its mechanism may be inhibiting the signaling pathway of NF-kB and MAPK to inhibit the production of the inflammatory factors of NO and IL-6.