异欧前胡素通过NF-κB和JAK1/STAT1信号通路抑制巨噬细胞M1极化发挥抗炎镇痛作用

Anti-inflammation and analgesic effect of isoimperatorin on the polarization of macrophage M1 via NF-κB and JAK1/STAT1 signaling pathway

目的探讨异欧前胡素抗炎镇痛作用的潜在药理作用机制。方法通过醋酸致小鼠扭体模型观察异欧前胡素的镇痛作用,采用二甲苯致小鼠耳肿胀模型观察抗炎作用;通过脂多糖(LPS,100 ng·mL^(-1))+干扰素-γ(IFN-γ,20 ng·mL^(-1))诱导小鼠巨噬细胞系RAW264.7构建M1极化模型,分别采用CCK8法、Real-time PCR法、ELISA法测定异欧前胡素对RAW264.7细胞增殖、M1极化标志蛋白(CD86和iNOS)的mRNA水平及白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)表达水平的调控作用;通过Western blot法检测NF-κB和JAK1/STAT1信号途径相关蛋白表达水平。结果异欧前胡素(30和60 mg·kg^(-1))能有效减少醋酸致扭体次数,抑制小鼠耳郭肿胀。异欧前胡素(20和40μmol·L^(-1))能抵抗LPS/IFN-γ诱导的RAW264.7细胞M1极化,抑制细胞增殖,降低IL-6和TNF-α的表达,并能有效地降低NF-κB和JAK1/STAT1信号途径相关蛋白磷酸化和转录。结论异欧前胡素可通过调节NF-κB和JAK1/STAT1信号通路抑制巨噬细胞M1极化,发挥抗炎镇痛作用。

Objective To determine the potential pharmacological mechanism of anti-inflammation and analgesic effect of isoimperatorin.Methods The anti-inflammation and analgesic effect of isoimperatorin was observed by acetic acid-induced writhing model and xylene induced ear swelling model in mice.M1 polarization model of mouse macrophage RAW264.7 was constructed by lipopolysaccharide(100 ng·mL^(-1))+IFN-γ(20 ng·mL^(-1)).The effect of isoimperatorin on the proliferation of macrophage,mRNA levels of CD86 and iNOS,and the protein level of IL-6 and TNF-αwas measured by CCK8,Real-time PCR and ELISA respectively.Finally,Western blot was used to determine the protein expression levels of NF-κB and JAK1/STAT1 signaling pathway.Results Isoimperatorin(30 and 60 mg·kg^(-1))decreased the number of body writhings,and attenuated ear swelling induced by xylene.Isoimperatorin(20 and 40μmol·L^(-1))inhibited the M1 polarization of RAW264.7 cells,decreased the expression of IL-6 and TNF-α,and decreased the phosphorylation of NF-κB and JAK1/STAT1 signaling pathway.Conclusion Isoimperatorin has significant anti-inflammation and analgesic effects,the mechanism may be related to inhibition of the M1 polarization of macrophages via NF-κB and JAK1/STAT1 signaling pathway.