复方鳖甲软肝片防治大鼠肝纤维化作用及机制

Effect and Mechanism of Fufang Biejia Ruangan Tablet in Prevention and Treatment of Rat Hepatic Fibrosis

目的:探讨复方鳖甲软肝片对大鼠肝纤维化的干预作用及机制。方法:SD成年大鼠随机分为空白对照组(A组)、复方鳖甲软肝片治疗组(B组)、复方鳖甲软肝片干预组(C组)和模型对照组(D组),B、C、D组大鼠采用CCl4诱导大鼠肝纤维化模型,B组于造模第6周给予复方鳖甲软肝片,C组在造模的同时给予复方鳖甲软肝片;于造模第6、12周检测大鼠血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、白蛋白(ALB)、总胆红素(TBIL)等肝功能指标,造模第12周处死大鼠检测肝脏指数,ELISA检测大鼠血清透明质酸、层黏连蛋白、Ⅲ型前胶原、Ⅳ型胶原等肝纤维化指标;提取各组大鼠肝脏Kupffer细胞,免疫荧光染色检测CD14表达,观察Kupffer细胞活化情况;实时荧光定量PCR及Western Blot分别检测Kupffer细胞中TGF-βmRNA及蛋白表达水平。结果:经复方鳖甲软肝片干预或治疗后,B、C组大鼠肝脏指数、肝纤维化指标、肝功能指标均优于D组,差异具有统计学意义(P <0. 05); D组Kupffer细胞中CD14呈阳性,B、C组CD14呈阴性; B、C组大鼠kupffer细胞中TGF-β蛋白及mRNA水平显著低于D组,差异具有统计学意义(P <0. 05)。结论:复方鳖甲软肝片具有抑制肝Kupffer细胞活化的作用,并可能通过抑制其分泌TGF-β而发挥抗肝纤维化的作用。

Objective:To investigate the preventive and therapeutic effects of Fufang Biejia Ruangan Tablet on CC14-induced hepatic fibrosis in rats and its action mode.Metliods:Sprague-Dawley adult rats were randomly divided into control group(group A),group B,group C and group D.Rats in groups B,C and D were treated with CC14to induce rat liver fibrosis model.Rats in group C were given with Fufang Biejia Ruangan Tablets at the sam e time as C1C4treatment.Rats in group B were given with Fufang Biejia Ruangan Tablets at6th week after C1C4treatment.At the6th and12th week of C1C4treatment,serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),album in(ALB)and total bilirubin(TBIL)were measured and the rats were sacrificed at the12th week.The liver fibrosis indexes such as serum hyaluronic acid,laminin,typeⅢprocollagen and typeⅣcollagen were detected by ELISA.CD14expression was detected by immunofluorescent staining in Kupffer cells isolated from the livers of each group to observe Kupffer cell activation;transforming growth factor(TGF-β)expression in Kupffer cells and livers was assessed by real-time quantitative PCR and Western Blot.Results:After treatment with Fufang Biejia Ruangan Tablets,the health status including the liver index,liver fibrosis index and liver function index of rats in group B and C was better than that of rats in group D,the difference was statistically significant(P<0.05);C D14expression w as positive in Kupffer cells in group D,but negative in group A,B and C.Moreover of TGF-βat both m R N A protein in kupffer cells in group B and C were significantly lower than group D(P<0.05).Taken together,anti-fibrosis effect of Fufang Biejia Ruangan Tablet might i m pairs TGF-βexpression and secretion by Kupffer cells via inhibiting the activation of kupffer cells.