益气化瘀化痰养阴法单剂与合剂对肝纤维化大鼠转化生长因子-β1受体表达的影响

Effects of TGF-β1R receptor expression in rats of liver fibrosis by single-dose and mixture of YiQi HuaYu HuaTan YangYin

目的观察益气化瘀化痰养阴法对CCl4诱导的大鼠肝纤维化肝组织转化生长因子(TGF)-β1受体表达的影响,探讨单剂与四法合剂对肝纤维化的疗效评价,以期更好指导中西医临床选择用药。方法将雄性SD大鼠随机分为正常对照组、肝纤维化模型组、益气组、化瘀组、化痰组、养阴组、益气化瘀化痰养阴组。除正常对照组外,各实验组采用皮下注射CCL4+低蛋白高脂饮食+酒精饮料的方法制备肝纤维化模型。模型成功后,根据组别分别灌服益气、化瘀、化痰、养阴单剂和与四药合剂。于12周末,根据组织检测要求分别进行心脏灌注和直接取大鼠肝脏等方法制备标本。通过Masson三色胶原染色、光镜以观察肝纤维化结构特点,采用RT-PCR及Western印迹方法观察各组TGF-β1受体的基因和蛋白表达变化。结果与模型组相比,益气、化瘀、化痰、养阴法均可以减轻肝组织内纤维化程度,降低大鼠肝组织TGF-β1受体基因和蛋白表达(P<0.05)。其中,益气组疗效最差(P<0.01),养阴法降低大鼠肝组织TGF-β1受体基因和蛋白表达更明显(P<0.05),化瘀和化痰两组之间无明显差异(P>0.05)。与单用相比,四种治疗方法合用更能显著降低大鼠肝组织TGF-β1受体基因和蛋白表达(P<0.05)。结论益气、化瘀、化痰、养阴法均可以降低肝组织内纤维化程度,降低TGF-β1受体的表达,虽说单剂有较好的疗效以及有不同差异,但是在四法合用中却能够发挥辅助作用,更有益于临床治疗。

Objective To observe effects of YiQi, HuaYu, HuaTan,YangYin method on TGF-β1 R receptor in rat models of hepatic fibrosis caused by CCl4 composed factor, to explore the therapeutic effect of hepatic fibrosis by single-dose and mixture of YiQi, HuaYu, Hua- Tan, YangYin. Methods The male SD rats were randomly divided into normal control, liver fibrosis model, YiQi, HuaYu, HuaTan and YangYin groups. Except the rats in normal control group, the rest rats of the experimental groups were made liver fibrosis model by subcutaneous injection of carbon tetraehloride ＋ low-protein high-fat diet ＋ alcohol. After the success of modeling, the experimental rats were randomly divided into appropriate groups, according to the group rats were fed single-dose and mixture of YiQi, HuaYu, HuaTan, YangYin. Liver tissue samples were collected and heart reperfusion was carried at 12 weeks of experiment. Hepatic fibrosis was observed by Masson staining and electron microscopic observation. RT-PCR and Western blot were used to examine TGF-β1 receptor gene and protein expression. Results Compared with model group, YiQi,HuaYu,HuaTan,YangYin method could reduce the degree of fibrosis in liver tissue, and reduce TGF-β1R receptor gene and protein expression （P 〈 0. 01 ）. Effect of YiQi was worst. Effect of YangYin method on reducing TGF-β1 R receptor gene and protein expression was more obvious （ P 〈 0. 05 ）. There was no difference between HuaTan and HuaYu groups （ P 〈 0. 05 ）. Compared with single-use, combination of four treatment could reduce TGF-β1R receptor gene and protein expression in rat liver tissue （P 〈 0. 05）. Conclusions YiQi, HuaYu, HuaTan,YangYin method can reduce the liver tissue fibrosis and reduce TGF-β1 receptor expression. Compared with single method, effect of combination of YiQi, HuaYu, HuaTau, YangYin methods is better.