肝纤维化不同证型与TGFβ1Smad基因蛋白表达关系实验研究.docVIP

肝纤维化不同证型与TGFβ1Smad基因蛋白表达关系实验研究 　　摘 要:目的:观察肝纤维化不同证型与TGF-β1/Smad基因蛋白的关系。方法:分别设立正常对照组、单纯肝纤维化组、肝纤维化肝郁脾虚证组和气虚血瘀证组,各模型组先用CCl?4注射法复制肝硬化疾病模型,然后肝郁脾虚证组附加夹尾和大黄灌胃法、气虚血瘀证组附加游泳疲劳法,并以相应药物干预。采用免疫组化法测定肝组织TGF-β1、TβR-Ⅰ、Smad2/3及Smad7水平,用比色法测定肝组织Smad3mRNA。结果:TGF-β1、TβR-Ⅰ、Smad2/3和Smad7在模型对照组中表达明显增强,以肝纤维化气虚血瘀组最为显著,肝纤维化肝郁脾虚组和单纯肝纤维化组的表达面积和强度相似。各模型组Smad3mRNA呈上调表达,差异均具有十分显著的意义(P1/Smad信号转导通路参与了肝纤维化的发生发展,肝纤维化不同证型之间TGF-β1/Smad基因蛋白表达有差异。 　　关键词:肝纤维化;TGF-β1/Smad基因蛋白;肝郁脾虚;气虚血瘀 　　中图分类号:R2-03 文献标识码:A 文章编号:1673-7717(2010)01-0023-06 　　 　　Experimental Study on the Relations between the TGF-β1/Smad and Different TCM Syndromes of Liver Fibrosis 　　XU Shan,BAO Jian?feng,ZHOU Min,ZHANG Yong?sheng 　　(Zhejiang Chinese Medical University,Hangzhou 310053,Zhejiang,China) 　　Abstract:Objective:To observe the relations between the TGF-β1/Smad and different TCM syndromes of liver fibrosis.Methods:All rats of model groups were treated by CCl?4 to establish the liver fibrosis models. Besides CCl?4, the rats of liver-stagnation and spleen-deficiency group were treated by stimulating tails with forceps clip and gastric infusion of rhubarb decoction, and the rats of qi-deficiency and blood-stasis group were treated by the method of swimming-fatigue. Then every group was intervened by the relevant medicine. After all treatments, the TGF-β1,TβR-Ⅰ,Smad2/3, Smad7 and Smad3mRNA of liver were examined.Results:The TGF-β1, TβR-Ⅰ, Smad2/3 , Smad7 and Smad3mRNA? of model groups were expressed significantly(P1/Smad participate in the occurrence and development of liver fibrosis.There are differences of TGF-β1/Smad expression among different TCM syndrome groups. 　　Key words:liver fibrosis;TGF-β1/Smad;liver-stagnation and spleen-deficiency;qi-deficiency and blood-stasis 　　 　　肝纤维化是各种慢性肝病的共同病理学基础,是发展至肝硬化的必经阶??,中医药在阻断和逆转肝纤维化方面取得了较好的效果,是防治肝硬化的研究热点。研究发现TGF-β1在肝纤维化的启动、进展乃至肝硬化的形成中发挥了重要作用[1-4],而近年Smad蛋白家族的发现,则从细胞内信号转导、细胞因子水平揭示了肝纤维化形成机制。因此研究TGF-β1/Smad基因蛋白的表达与肝纤维化及肝纤维化中医辨证分型的关系,将为中医治疗肝纤维化的作用机制和靶点提供实验依据。 　　 　　1 材料与方法 　　 　　1.1 动物及