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目的观察没食子酸对脂多糖(LPS)诱导RAW264.7巨噬细胞Toll样受体4/核因子κB(TLR4/NF-κB)通路的影响。方法将巨噬细胞分为空白对照组、LPS组、LPS联合没食子酸组、LPS联合NF-κB抑制剂吡咯二硫代甲酸(PDTC)组和LPS联合地塞米松(DM)组。处理后的细胞培养24 h,ELISA检测肿瘤坏死因子α(TNF-α)、白细胞介素1(IL-1)、IL-6水平,实时定量PCR检测TLR4、NF-κB mNRA水平,Western blot法检测p65、p-p65、TLR4、磷酸化的NF-κB抑制蛋白α(IκBα)的蛋白表达水平。结果 LPS诱导RAW264.7巨噬细胞后TNF-α、IL-1、IL-6水平升高,没食子酸可降低LPS诱导引起的TNF-α、IL-1和IL-6表达水平升高。LPS刺激后TLR4 mRNA及蛋白表达增加,NF-κB活化,没食子酸可拮抗以上作用,阻止NF-κB活化。结论没食子酸可通过TLR4/NF-κB通路抑制LPS诱导的RAW264.7巨噬细胞炎性反应。
Objective To investigate the effect of gallic acid on TLR4 / NF-κB pathway induced by lipopolysaccharide (LPS) in RAW264.7 macrophages. Methods Macrophages were divided into blank control group, LPS group, LPS combined with gallic acid group, LPS combined with PDTC group and LPS plus dexamethasone group. The cultured cells were cultured for 24 hours. The levels of TNF-α, IL-1 and IL-6 were detected by ELISA. The levels of TLR4 and NF-κB mNRA were detected by real-time quantitative PCR. The protein expression levels of p65, p-p65, TLR4 and phosphorylated NF-κB inhibitor α (IκBα) were detected. Results LPS induced the increase of TNF-α, IL-1 and IL-6 levels in RAW264.7 macrophages. Gallic acid decreased the expression of TNF-α, IL-1 and IL-6 induced by LPS. LPS stimulated TLR4 mRNA and protein expression, NF-κB activation, gallic acid can antagonize the above effect, prevent NF-κB activation. Conclusion Gallic acid can inhibit LPS-induced RAW264.7 macrophage inflammatory response through TLR4 / NF-κB pathway.