柴胡皂甙D对肺纤维化小鼠上皮-间质转化的干预作用及机制研究

Intervention effect and mechanism of saikosaponin D on epithelium-mesenchymal transformation in mouse with pulmonary fibrosis

目的观察柴胡皂甙D(SSD)对肺纤维化小鼠上皮-间质转化(EMT)的影响及其机制。方法将60只雄性昆明小鼠分为对照组、博来霉素(BLM)组和SSD组,BLM组和SSD组通过气管内注入浓度为5mg/kg的BLM溶液制备肺纤维化模型,对照组给予相同体积的0.9%氯化钠。SSD组每日腹腔内注入SSD溶液(2.0mg/kg),BLM组和对照组每日同等条件下腹腔内注入0.2mL生理盐水+DMSO(3mL DMSO溶于97mL生理盐水)。分别在第14天和第28天,处死小鼠取肺组织行病理苏木素-伊红(HE)染色,观察肺泡炎和肺纤维化程度,并通过Western blot检测肺组织E钙黏蛋白、纤维连接蛋白、Wnt蛋白和β-actenin蛋白的表达水平。结果 SSD组和BLM组肺泡炎及肺纤维化程度评分均高于同期对照组,且SSD组低于同期BLM组;SSD组和BLM组纤维连接蛋白、Wnt蛋白和β-actenin蛋白表达均高于同期对照组,且SSD组低于同期BLM组;SSD组和BLM组E钙黏蛋白表达均低于对照组,且SSD组高于同期BLM组。结论 SSD对肺纤维化有一定治疗价值,SSD抑制Wnt/β-catenin信号传导通路负向调节EMT过程可能是其治疗机制之一。

Objective To observe the effect and mechanism of saikosaponin D（SSD）on epithelium mesenchymal transformation（EMT）in the mouse with pulmonary fibrosis.Methods Sixty male Kunming mice were divided into the control group,bleomycin（BLM）group and SSD group.The pulmonary fibrosis model in the BLM group and SSD group was prepared by intratracheal instillation of 5 mg/kg BLM solution and the control group received the same volume of 0.9% sodium chloride solution.The SSD group was daily intraperitoneally injected by 2.0 mg/kg SSD solution,and 0.2 mL normal saline plus DMSO（3 mL DMSO was dissolved in 97 mL normal saline）were daily intraperitoneally injected on the equal conditions in the control and BLM group.The mice were executed on 14,28 drespectively for taking the pulmonary tissues.Then the HE staining was performed to observe alveolitis and fibrosis degree in lung tissues.Western blot was performed to determine the expression level of E-cadherin,fibronectin,Wnt protein andβ-actenin protein in lung tissues.Results The scores of alveolitis and pulmonary fibrosis in the SSD group and BLM group were higher than those in the control group at the corresponding periods,moreover the SSD group was lower than the BLM group at the corresponding periods.The relative gray values of fibronectin,Wnt protein andβ-actenin protein expression in the SSD group and BLM group were higher than those in the control group at the corresponding periods,moreover the SSD group was lower than the BLM group at the corresponding periods;the relative gray value of the expression of E-cadherin in the SSD group and BLM group was lower than that in the control group at the corresponding periods,moreover the SSD group was higher than the BLM group at the corresponding periods.Conclusion SSD has a certain treatment value for pulmonary fibrosis,and SSD inhibiting the Wnt/β-catenin signal transduction pathway for negatively regulating EMT process may be one of its treatment mechanisms.