金钗石斛多糖对肝纤维化大鼠TGF-β1、α-SMA、Ⅰ型胶原、Ⅲ型胶原表达的影响

Effect of dendrobium nobile polysaccharides on the expression of TGF-β1, α-SMA, type Ⅰ,Ⅲ collagen in rats with hepatic fibrosis

目的探讨金钗石斛多糖(DNP)对肝纤维化(HF)大鼠肝组织转化生长因子(TGF)-β1、α-平滑肌肌动蛋白(α-SMA)、Ⅰ、Ⅲ型胶原表达的影响。方法SD大鼠随机分为正常组,模型组,秋水仙碱组(0.2 mg/kg),扶正化瘀组(0.415 g/kg),DNP低、中、高剂量组(5、10、20 g/kg),采用50%四氯化碳腹腔注射和30%乙醇灌胃诱导大鼠HF模型,造模8 w后,给予相应药物(10 ml/kg)进行灌胃,每日1次,给药4 w,正常组、模型组灌胃生理盐水。苏木素-伊红(HE)染色和Masson染色观察HF程度;免疫组化法检测肝组织TGF-β1、α-SMA、Ⅰ、Ⅲ型胶原蛋白表达;荧光定量聚合酶链反应(PCR)检测肝组织Ⅰ、Ⅲ型胶原mRNA表达。结果与正常组比较,模型组明显纤维化(P<0.01),TGF-β1、α-SMA、Ⅰ、Ⅲ型胶原蛋白表达明显升高(P<0.01),Ⅰ、Ⅲ型胶原mRNA表达亦明显升高(P<0.01);与模型组比较,DNP各组肝纤维化程度均有所减轻(P<0.01),DNP降低TGF-β1、α-SMA、Ⅰ、Ⅲ型胶原蛋白表达(P<0.01),DNP降低Ⅰ、Ⅲ型胶原mRNA表达(P<0.01)。结论DNP有抗纤维化作用,其机制可能与下调TGF-β1、α-SMA、Ⅰ、Ⅲ胶原蛋白表达,降低Ⅰ、Ⅲ胶原mRNA表达有关。

Objective To explore the effect of Dendrobium nobile polysaccharide(DNP)on the expression of transforming growth factor(TGF)-β1,α-smooth muscle actin(α-SMA),and typeⅠ,Ⅲcollagen in liver tissues of rats with hepatic fibrosis(HF).Methods SD rats were randomly divided into normal group,model group,colchicine group(0.2 mg/kg),Fuzheng Huayu group(0.415 g/kg),DNP low,medium and high dose groups(5,10,20 g/kg),50%carbon tetrachloride intraperitoneal injection and 30%ethanol gavage were used to induce HF model in rats,8 weeks after modeling,corresponding drugs(10 ml/kg)gavage administration once a day for 4 weeks.The normal group and the model group were given normal saline.HE staining and Masson staining were used to observe the degree of liver fibrosis;immunohistochemical method was used to detect liver tissue TGF-β1,α-SMA,Ⅰ,Ⅲcollagen expression;fluorescence quantitative PCR method was used to detect liver tissue typeⅠandⅢcollagen mRNA expression.Results Compared with the normal group,the model group was significantly fibrotic(P<0.01),the expressions of TGF-β1,α-SMA,typeⅠandⅢcollagen were significantly increased(P<0.01),and the expressions of typeⅠandⅢcollagen mRNA were also significantly increased(P<0.01).Compared with the model group,the degree of liver fibrosis in each group of DNP was alleviated(P<0.01),the expressions of TGF-β1,α-SMA,typeⅠandⅢcollagen were significantly decreased(P<0.01),and the expressions of typeⅠandⅢcollagen mRNA were also significantly decreased(P<0.01).Conclusions DNP has a good anti-fibrosis effect,and its mechanism may be related to the down-regulation of the expression of TGF-β1,α-SMA,Ⅰ,Ⅲcollagen and the decrease ofⅠandⅢcollagen mRNA expression.