经方小柴胡汤对小鼠肝纤维化的改善作用及其机制的研究

Study on Effect and Mechanism of Xiaochaihu Decoction Ameliorating Hepatic Fibrosis

目的:探究经方小柴胡汤对肝纤维化改善作用的机制。方法:将雄性C57BL/6小鼠随机分为3组,即正常对照组、模型组和实验组。模型组和实验组小鼠腹腔注射二甲基亚硝胺(DMN),每周3次,实验组注射DMN的同时每天灌胃小柴胡汤。4周后麻醉处死各组小鼠,取小鼠血浆和肝脏,对肝脏进行HE和masson染色;利用ELISA检测试剂盒测定血清中白细胞介素1β(IL-1β)、白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)和C反应蛋白(CRP)以及肝脏4-羟脯氨酸(4-Hyp)含量;采用realtime PCR和western blot检测肝脏IL-1β、IL-6、TNF-α、I型胶原蛋白(Collagen I)和α-平滑肌动蛋白α(α-SMA)表达量、核因子κB抑制蛋白(IκB)的磷酸化水平及核因子κB(NF-κB)核积累量。结果:结果表明,实验组肝纤维化病理变化相比于模型组得到显著改善(P<0.05);实验组血清IL-1β、IL-6、TNF-α、CRP及肝脏4-Hyp与模型组相比其含量均明显下降,且差异显著(P<0.05);实验组肝脏IL-1β、IL-6、TNF-α、Collagen Ⅰ和α-SMA的表达水平、IκB的磷酸化水平及NF-κB的核积累量相比于模型组显著下降(P<0.05)。结论:小柴胡汤能显著改善小鼠肝纤维化,其作用机制与抑制肝脏内的炎症反应有关。

Objective： To understand the mechanism of the classical prescription Xiaochaihu Decoction ameliorating hepatic fibrosis. cleus accumulation of nuclear factor KB （ NF - KB ） were measured by reahime PCR and western blot. Results ： The path- ological change ofMethods： Male C57BL/6 mice were divided into three groups in this experiment, namely normal control, model control and experiment group, and dimethylnitrosamine （DMN） was injected into the mice of the model and experiment groups three time per week intraperitoneally. The mice in experiment group were given garage of Xiaochaihu Decoction once a day simultaneously. Four weeks later, the mice sacrificed, and the plasm was collected and the livers were removed. HE and masson staining of livers were carried out. The cytokines, C -reactionprotein （CRP） levels in serum and the 4 - hydroxyproline level in livers were measured by ELISA. The expressions of IL - 1β, IL - 6, TNF -α, Collagen I and α - smooth muscle actin （ α - SMA）, the phosphorylation level of inhibitor of nuclear factor κB （κB） and the nu- hepatic fibrosis of experiment group was improved significantly compared with that of the model group （P 〈 0. 05 ）. The contents of serum IL - 1β, IL - 6, TNF - α, CRP and the liver 4 - Hyp of the experiment group decreased significantly compared with those of the model group （ P 〈 0. 05）. The expressions of IL - 1β,IL - 6, TNF -α, Collagen I and α - SMA, the phosphorylation level of κB and the nucleus accumulation of NF - κB of the experiment group were down - regulated significantly compared with those of the model group （ P 〈 0. 05 ）. Conclusion ： Xiaochaihu Decoction can ameliorate hepatic fibrosis and the mechanism of its action is associated with inhibiting inflammation.