扶正化瘀方促进二甲基亚硝胺肝纤维化大鼠肝窦毛细血管化逆转作用的实验研究

Effect of Fu Zheng Hua Yu Recipe on hepatic sinusoid capillarization in the rat of dimethyl nitrosamine-induced hepatic fibrosis

目的:探讨临床有效的抗肝纤维化扶正化瘀319方(下称319方)对肝纤维化、肝硬化常见病理变化肝窦毛细血管化的逆转作用。方法:采用二甲基亚硝胺(DMN)4周12次腹腔注射制备大鼠肝纤维化模型。造模成型后分为模型对照组与319方治疗组,用药4周;获取血清与肝组织,作病理组织学、电镜超微结构、免疫组化、肝组织羟脯氨酸含量及肝功能观测。结果:正常组大鼠肝窦内皮细胞外周胞浆很薄,有许多小窗孔,内皮下未见基底膜。模型组大鼠肝窦扭曲、狭窄,内皮细胞胞浆窗孔减少或消失,有明显的基底膜出现;肝组织可见出血性坏死,门脉区大量的结缔组织增生,纤维间隔向小叶内伸展,肝组织Hyp含量显著增高;肝窦壁第Ⅷ因子相关抗原、α-平滑肌肌动蛋白、LM及Ⅳ型胶原阳性染色较正常大鼠均显著增强,且范围较广。与模型对照组比较,319方治疗组肝窦扭曲、狭窄程度及内皮窗孔减少和消失程度轻,基底膜较薄或不连续,部分肝窦结构接近正常;肝细胞变性坏死程度轻,肝组织内纤维间隔明显减少,Hyp含量显著降低(P<0.05);肝窦壁第Ⅷ因子相关抗原、α-平滑肌肌动蛋白、LM及Ⅳ型胶原阳性染色均显著减弱,分布面积图像分析结果均显著降低(P<0.05)。结论:扶正化瘀319方可显著促进肝纤维化、肝硬化常见病理变化肝窦毛细血管化的逆转。

Objective: To investigate the effect of Fu Zheng Hua Yu-319 Recipe on hepatic sinusoid capillarization in the rat of hepatic fibrogenesis. Methods: Hepatic fibrosis model rats were induced by intraperitoneal injection of dimethyl nitrosamine (DMN). thrice a week for 4 weeks, and they were divided into the treatment group which were treated with Fu Zheng Hua Yu-319 Recipe for 4 weeks, and the model control group. Serum and hepatic tissue were taken for examination of pathohistology, microstructure by electron microscopy, immunohistochemistry, and hydroxyproline content in the liver tissue and hepatic function. Results: In the normal group, peripheral cytoplasma of endothelial cells in the hepatic sinusoid is very thin with a lot of small fenestra, and no basement membrane under endothelium was found. In the model group, crookedness and stricture of the hepatic sinusoid were found , and the fenestra in cytoplasma of the endothelial cell reduced or disappeared with obvious basement membrane, and hemorrhage necrosis in the liver tissue could be found, with a great proliferation of connective tissue in the portal area, and fibrous septum extended towards into the lobuli, the content of hydroxyproline (HYP) in the liver tissue increased significantly. The VI factor-related antigen, a-actin of smooth muscle, LM and type IV collagen positive staining on the hepatic sinusoid wall increased significantly with a wider range as compared with the normal rat. In the treatment group, the extents of crookedness and stricture of hepatic sinusoid and reduction and disappearence of endothelial fenestra were lower, with thiner or discontinuous basement membrane, and a part of hepatic sinusoid construction approached to normal, and degeneration and necrosis of hepatic cells were mild and fibrous septum in the liver tissue decreased significantly, and HYP content decreased significantly (P < 0.05), and the VI factor-related antigen, a-actin of smooth muscle, LM and type IV collagen positive staining on the hepatic sinusoid walls weakened significantly, with significant decrease in the image analysis results of distribution area (P<0. 05). Conclusion: The Fu Zheng Hua Yu Recipe can effectively inhibit experimental hepatic fibrosis and sinusoid capillarization.