消黄方对二甲基亚硝胺致肝纤维化大鼠肝脏巨噬细胞及促炎症因子的干预作用

目的: 观察消黄方对二甲基亚硝胺(DMN)大鼠肝纤维化模型肝纤维化过程中肝脏巨噬细胞及促炎相关因子的干预作用。 方法: 每周3 d连续腹腔注射DMN(10 mg·kg^-1),4周后制备大鼠肝纤维化模型。造模2周末将DMN大鼠随机分为模型组和消黄方组(每组10只),造模第3周起消黄方组大鼠造模的同时每天给予消黄方1.73g·kg^-1,灌胃至4周末,模型大鼠每天给予双蒸水。留取肝组织样本及血清,免疫印迹和免疫组化及实时定量PCR检测巨噬细胞标志物CD68及肿瘤坏死因子(TNF-α),白介素1β (IL-1β)表达。 结果: 免疫组化结果显示,正常组CD68主要分布在肝窦,小叶间也有零星分布,DMN造模后CD68呈强阳性表达;实时定量PCR结果显示,与正常组相比,DMN造模4周CD68表达显著显著升高(P<0.01);肝脏TNF-α和IL-1β mRNA与CD68表达一致。与模型相比,消黄方显著抑制CD68及TNF-α及IL-β mRNA表达(P<0.01或P<0.01))。 结论: 消黄方对炎症反应的抑制作用可能其是发挥抗DMN肝纤维化效应机制之一。

Inhibition Macrophages Activation and Pro-inflammatory Cytokines of Xiaohuang Decoction in Dimethylnitrosamine-induced Liver Fibrosis in Rats

Objective:To investigate effects of Xiaohuang decoction on macrophages activation and pro-inflammatory cytokines in rats with dimethylnitrosamine(DMN)-induced liver fibrosis.Method: In DMN-treated rats,DMN(10 mg · kg-1) was administrated intraperitoneally for the first three days per week for 4 weeks.At the second weekend,DMN rats were randomly divided into DMN-water and DMN-Xiaohuang decoction(DMN-XHD).At the first day of the third week,both groups continued to receive weekly DMN treatment for another 2 weeks in addition to daily administration of either water or XHD.CD68 was detected using western blotting,immunohistochemistry and real-time PCR.Result:The immunohistochemistry results showed that CD68 mainly located in the sinusoids and between interlobular in normal group.The CD68 mainly located in the fibrotic septa in DMN-treatment rats.The CD68 expression was increased 5-fold compared with normal rats showed by real-time PCR analysis.Compared with DMN-water rats,CD68 was reduced significantly in XHD rats.The content of Tumor necrosis factor(TNF-α) in the serum increased remarkably in DMN-water rats,and XHD could suppress TNF-α notably.Conclusion: Xiaohuang decoction inhibited liver macrophage activation and pro-inflammatory cytokines in DMN-induced liver fibrosis.