常山酮对大鼠肝纤维化的影响及其机制

Effects of halofuginone on hepatic fibrosis in rats and their mechanism

目的探讨常山酮对刀豆蛋白A(Con A)诱导的大鼠肝纤维化的影响及其机制。方法 50只Wistar大鼠随机分成对照组(8只)、模型组(14只)及常山酮低、高剂量(5、10 mg/kg)组(各14只),对照组大鼠每周尾iv磷酸盐缓冲液(PBS)300μL 1次;其余3组大鼠每周尾iv Con A12.5 mg/kg(溶于300μL PBS)1次,连续8周。自造模起在常山酮各组大鼠饲料中添加相应剂量的常山酮,连续给药8周后大鼠称质量,处死。生化分析法检测血清中肝功能指标丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、总蛋白(TP)、白蛋白(ALB)的水平;ELISA法检测血清肝纤维化指标转化生长因子-β1(TGF-β1)、透明质酸(HA)、Ⅲ型前胶原(PC-Ⅲ)的水平;肝组织行HE和Masson胶原染色,进行病理分析并按肝纤维化半定量计分系统(SSS)计分系统进行评分;天狼星红和免疫组化染色观察肝组织Ⅰ型胶原(Col Ⅰ)的沉积及蛋白表达的变化。结果与模型组大鼠比较,常山酮组大鼠血清中ALT、AST、TP、TGF-β1、HA和PC-Ⅲ水平显著降低(P<0.05、0.01),肝脏病理损伤明显减轻(P<0.05、0.01),肝纤维化评分明显降低(P<0.05、0.01),肝组织中Col Ⅰ沉积及蛋白的表达显著下调(P<0.05、0.01)。结论常山酮能有效减轻ConA致肝纤维化大鼠的肝脏损伤,降低肝纤维化程度,其机制与抑制肝内Col Ⅰ的沉积和蛋白的表达有关。

Objective To investagate the protective effects of halofuginone against liver injury induced by eoncanavalin A （Con A） in rats and their mechanism. Methods Rats were divided into control （G1, 8）, model （G2, 14）, low- （5 mg/kg, G3, 14） and high-dose （10 mg/kg, G4, 14） halofuginone groups. The rats in G1 were iv injected with 300 p,L PBS from tail vain once a week, the rats in other three groups were iv injected with 12.5 mg/kg Con A （in 300 ~tL PBS） once a week for eight consecutive weeks. Halofuginone was given in the diet for rats in G3 and G4 after molding and continuous administration for eight weeks after the rats were weighted and then were put to execution. The biochemical analysis was used to determine alanine aminotransferase （ALT）, aspartate aminotransferase （AST）, total protein （TP）, and album protain （ALB） in serum while enzyme-linked immuno sorbent assay （ELISA） was applied for detecting transforming growth factor -β1 （TGF-β1）, hyaluronic acid （HA）, and procollagen III （PC-III） levels in serum. HE and Masson＇s trichrome stainings were conducted in liver tissue to observe the pathological variations. Grades of hepatic fibrosis were evaluated according to SSS system. Sirus and immunohistochemical stainings were executed for detecting the deposition and protein expression of collagen 1 （Col 1） in liver tissue. Results Compared with the G2, halofuginone could decrease the levels of ALT, AST, TP, ALB, TGF-β1, HA, and PC-Ill in serum obviously （P 〈 0.05, 0.01）. Halofuginone could improve the liver pathological variations of fibrotic rats obviously （P 〈 0.05, 0.01） and reduce the score of hepatic fibrosis significantly （P 〈 0.05, 0.01）. Halofuginone treatment could reduce the deposition and expression of Col I protein （P 〈 0.05, 0.01）. Conclusion Halofuginone could attenuate the Con A-induced immunological liver injury and the fibrosis level in rats. The mechanisms possibly contribute to down-regulating the deposition and expression of Col I protein in liver of rats.