参七内金散对四氯化碳肝纤维化模型大鼠肝星状细胞活化与凋亡的影响

目的：观察参七内金散抗大鼠肝纤维化的作用机制。方法：雄性SD大鼠30只随机分为3组：正常组、模型对照组、参七内金散组。以CCl4皮下注射造模,首次注射100%CCl45mL/kg体重,之后40%CCl4 橄榄油溶液3mL/kg皮下注射,2次/周,连续注射6周。造模成功后以0.8g/kg的参七内金散灌胃模型大鼠共2周,正常组与模型对照组则予以等量生理盐水灌胃,共2周。以盐酸水解法测定肝组织羟脯氨酸含量,分别以HE、天狼猩红染色观察肝组织的炎症与纤维化病理。免疫印迹及免疫组化检测α-SMA的表达,观察肝星状细胞活化;连续切片,依次TUNEL染色与α-SMA免疫组化双染色,观察肝星状细胞凋亡。结果：与正常大鼠比较,模型大鼠肝组织可见大量肝组织脂肪变性及胶原病理沉积,并可见纤维间隔形成;参七内金散显著能改善模型大鼠肝组织脂肪变性与胶原病理沉积,降低肝组织Hyp含量。正常大鼠仅表达少量α-SMA,主要分布于血管壁,未见凋亡星状细胞;模型组大鼠可见大量α-SMA表达,可见少量凋亡星状细胞;与模型组比较参七内金散组肝组织α-SMA表达显著减少（P≤0.01）,但凋亡肝星状细胞显著增多。结论：参七内金散具有抗肝纤维化作用;抑制肝星状细胞活化、促进活化的肝星状细胞凋亡是参七内金散抗肝纤维化的机理之一。

Efects of Shenqi Neijin Decoction on HSC Activation and Apoptosis in Rats with CCI4 -induced Liver Fibrosis

Objective：To investigate anti-fibrotic pharmacological actions of the Shenqi Neijin Decoction （SQNJ） in rats. Methods ：The liver fibrosis models were induced by injection of carbon tetrachloride （CC14） subcutaneously for 4 weeks in rats and then were divided into model control and treated groups while normal rats were used as normal control. After model establishment, the rats of treated groups were administered with SQNJ with a dosage of 0.8g/（ kg d） body weight by gavage for two weeks. The normal and model groups took the same volume of saline. The rat liver tissues were examined. Hepatic inflammation was stained with HE and collagen deposition with Sirius red. Hydroxyproline （Hyp） con- tent of liver was assayed with hydrochloric acid hydrolysis. The liver protein expressions of α - SMA were determined by immunohistochemistry staining and Western blotting respectively. Apoptotic of the HSC in rat liver tissues were determined by dual staining both of the terminal deoxynucleotidy transferrase UTP - nick end labeling （TUNEL） and of α - SMA im- munohistochemistry. Results： Model rats had significant hepatic fatty degeneration and collagen accumulation and fibrosis at liver. While SQNJ treated group showed slighter hepatic fatty degeneration and collagen deposition, and lower levels of Hyp content than those of the model. The α - SMA expression in hepatic tissues of normal rats was weak, but its expres- sion in the hepatic tissues of model rats increased significantly, and SQNJ treated group reduced the expression of α - SMA in hepatic tissues. The treatment of SQNJ induced remarkable increases in TUNEL - and α- SMA dual - positive ceils in 72h. Conclusion ： Shenqi Neijin Decoction has a good effect against liver fibrosis in rats and this action is associat- ed with inducing hepatic stellate cells apoptosis and inhibiting the activation of HSCs.