德昂酸茶水提物对小鼠RAW264.7巨噬细胞抗炎作用的研究

Study on Anti-inflammatory of Aqueous Extract of De’ang Pickled Tea on Murine RAW264.7 Macrophages

【目的】探究德昂酸茶水提物对小鼠RAW264.7巨噬细胞的抗炎作用与机制。【方法】以脂多糖(lipopolysaccharides,LPS)和干扰素-γ(interferon-gamma,IFN-γ)诱导的小鼠RAW264.7巨噬细胞为炎症模型,采用Griess试剂盒检测一氧化氮(nitric oxide,NO)含量,采用实时荧光定量PCR分析炎症基因诱导型一氧化氮合成酶(inducible nitric oxide synthase,INOS)、环氧合酶2(cyclooxygenase 2,COX2)、白细胞介素-1β(interleukin-1β,IL1β)、白细胞介素-6(interleukin-6,IL6)和核转录因子-κB(nuclear factor-κB,NF-κB)信号通路关键基因的mRNA表达水平,采用蛋白质免疫印迹检测NF-κB信号通路活化水平。【结果】与LPS/IFN-γ诱导炎症组相比,德昂酸茶水提物可显著降低NO含量以及INOS、COX2、IL1β和IL6的mRNA表达水平(P<0.05),显著增加NF-κB信号通路负调控因子NFKBIA和TNFAIP3的m RNA表达水平(P<0.05),显著抑制NF-κB信号通路关键上游激酶IKKα蛋白磷酸化和抑制蛋白IκBα磷酸化(P<0.05)。【结论】德昂酸茶水提物可以通过调节NF-κB信号通路基因表达、关键上游激酶活化和抑制蛋白磷酸化从而抑制炎症基因表达,进而抑制巨噬细胞炎症反应。

[Purpose]To explore the anti-inflammatory effect and mechanism of aqueous extract of De’ang pickled tea on murine RAW264.7 macrophages.[Methods]Murine RAW264.7 macrophages induced with lipopolysaccharides(LPS)and interferon-gamma(IFN-γ)were used as inflammatory model.Content of nitric oxide(NO)was detected with the Griess kit,the mRNA expression levels of inducible nitric oxide synthase(INOS),cyclooxygenase 2(COX2),interleukin-1β(IL1β),interleukin-6(IL6),and key genes involved in nuclear factor-κB(NF-κB)signaling pathway were analyzed by real-time fluorescence quantification PCR.Activation NF-κB signaling pathway was detec-ted with Western-blotting.[Results]Compared with the inflammatory group,aqueous extract of De’ang pickled tea significantly decreased NO content and mRNA expression levels of INOS,COX2,IL1βand IL6(P<0.05),while significantly increased the mRNA expression levels of negative regulat-ory factors NFKBIA and TNFAIP3 in NF-κB signaling pathway(P<0.05),and significantly inhibited the phosphorylation of key upstream kinase IKKαand inhibitory protein IκBαof NF-κB signaling pathway(P<0.05).[Conclusion]Aqueous extract of De’ang pickled tea inhibits macrophage in-flammation through regulating the expression of NF-κB signaling pathway genes and activation of key upstream kinases and phosphorylation of inhibitory proteins as wells as inflammatory gene expression.