矮地茶黄酮对肝纤维化大鼠肝脏毛细血管异常增生的影响

The Effect of the Flavones from Ardisia Japonica on the Angiogenesis in Hepatic Fibrosis of Rats

目的:探讨矮地茶黄酮防治肝纤维化的作用及机制。方法:将SD大鼠随机分为5组(正常组、模型组、矮地茶黄酮高中低三个剂量组),腹腔注射二甲基亚硝胺制备大鼠肝纤维化模型,矮地茶黄酮组按200、400、600 mg/(kg·d)三种剂量灌胃进行干预。实验结束后,采集大鼠血液及肝组织标本,酶联免疫吸附试验法(ELISA)检测各组大鼠血清转化生长因子β1(transforming growth factor beta 1,TGF-β1)水平,HE染色观察各组大鼠肝脏炎症程度和纤维化程度,免疫组化方法检测各组大鼠肝组织中CD34、TGF-β1、碱性成纤维细胞生长因子(basic fibro⁃blast growth factor,b-FGF)的表达情况并进行相关性分析。结果:模型组大鼠肝脏中毛细血管数量增多且与肝纤维化程度呈正相关(均P<0.05),血清中TGF-β1水平、肝脏中TGF-β1、b-FGF的表达增高且与毛细血管数量均呈正相关(均P<0.05);与模型组相比,矮地茶黄酮高、中剂量组大鼠肝脏炎症程度、纤维化程度、新生毛细血管数量、血清中TGF-β1水平、肝脏中TGF-β1、b-FGF的表达明显降低(均P<0.05)。结论:矮地茶黄酮具有防治肝纤维化的作用且与剂量有关,作用机制可能与降低TGF-β1和b-FGF的表达,进而影响肝星状细胞增殖,抑制血管内皮细胞增生,减少血管生成有关。

Objective The purpose of this study was to investigate the effect and mechanism of the flavones from Ardisia ja⁃ponica on hepatic fibrosis(HF)of rats.Methods SD rats were randomly divided into five groups,normal group,model group,low dose group,middle dose group and high dose group.The liver fibrosis models were introduced by intraperitoneal injection of dimethylnitrosamine.The rats in the Aidicha flavone group were intragastrically perfused with flavones 200,400 and 600 mg/(kg·d).At the end of the experiment,the blood and liver tissue samples were collected,and the serum transforming growth factor beta 1(TGF-β1)was detected by enzyme-linked immunosorbent assay(ELISA).HE staining was used to observe the degree of liver inflammation and fibrosis,and immunohistochemistry was used to detect the the ex⁃pression of CD34,TGF-β1 and basic fibroblast growth factor(b-FGF)in rat liver tissues.The correlation was analyzed.Results In the model group,the number of capillaries in the liver was increased and positively correlated with the degree of liver fibrosis(all P<0.05).The levels of TGF-β1 in the serum and TGF-β1 and b-FGF in the liver were increased and positively correlated with the number of capillaries(all P<0.05).Compared with the model group,the degree of liver in⁃flammation,fibrosis,the number of new capillaries,the level of TGF-β1 in serum and the levels of TGF-β1 and b-FGF in the liver in the high and medium dose groups were significantly decreased(all P<0.05).Conclusion The flavonoids from Ardisia japonica have the dose-related effect of preventing and treating liver fibrosis.The mechanism of the action may be related to reducing the expression of TGF-β1 and b-FGF,thereby affecting the proliferation of hepatic stellate cells,in⁃hibiting the proliferation of vascular endothelial cells,and reducing angiogenesis.