Effects of Xiaohuang decoction on anti-oxidative stress and macrophage activation in lungs of rats with DMN-induced liver fibrosis
目的观察消黄方对二甲基亚硝胺(DMN)诱导的肝纤维化过程中大鼠肺组织氧化应激及巨噬细胞激活的干预作用。方法腹腔注射DMN 4周制备大鼠肝纤维化模型,4周末处死全部大鼠。检测肺组织超氧化物歧化酶(SOD)活性,谷胱甘肽(GSH)及丙二醛(MDA)含量;免疫印迹、免疫组化及实时定量PCR检测CD68表达。结果肺组织SOD活性及GSH含量在DMN染毒2周即降至较低水平,染毒4周维持低水平状态。MDA含量在DMN染毒2周到达较高水平,消黄方显著降低MDA含量并显著升高SOD活性及GSH含量。免疫组化显示正常组CD68在肺泡周呈弱阳性表达,DMN染毒过程中,CD68阳染进行性增强,消黄方组CD68阳染较弱。结论 DMN诱导肝纤维化过程中肺组织氧化应激和巨噬细胞激活紊乱。消黄方显著抑制肺组织氧化应激及巨噬细胞激活。Abstract:
Objective To observe the effects of Xiaohuang decoction (XHD) on anti-oxidative stress and macrophage activation in lungs of rats with dimethylnitrosamine (DMN) -induced liver fibrosis.Methods Liver fibrosis was induced in male Wistar rats by DMN injection, three days a week for four weeks and divided into two groups: untreated model group and XHD-treated group.XHD or distilled water was administered eight times a day for two weeks.All rats were sacrificed at the end of the 4th week and lung tissues harvested for analysis.Oxidative stress indicators [superoxide dismutase (SOD) , glutathione (GSH) , and malondialdehyde (MDA) ] in the lung were detected by biochemical assays.CD68 expression in the lung was detected by Western blot, immunohistochemistry, and real-time quantitative PCR.Results In the DMN-induced rats, SOD activity and GSH content had decreased remarkably after two weeks and was maintained at a low level state out to four weeks, but MDA content had increased after two weeks.XHD treatment significantly reduced the DMN-enhanced MDA content and notably increased the DMN-inhibited SOD activity and GSH content after four weeks.Immunohistochemical analysis of uninduced normal control rats revealed that CD68 was localized to the alveoli and weakly expressed.However, in DMN lung, the CD68 expression was strong after two weeks.XHD-treated DMN rats had weak CD68-positive staining in lung.Conclusion Oxidative stress and activation of macrophages occurred in the lung during DMN-induced hepatic fibrosis.XHD treatment inhibits oxidative stress and lung macrophage activation.
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