Shugan Huoxue Huayu Fang(疏肝活血化瘀方)attenuates carbon tetrachloride-induced hepatic fibrosis in rats by inhibiting transforming growth factor-β1/Smad signaling

• 作者: LIU Lei GUO Hanbin SHAO Cuiping WANG Lin XU Youqing ZHOU Yiming
• 来源期刊: 中医杂志（英文版）
• 年，卷（期）: 2022,（1）
• 所属期刊栏目: Research Articles
• 页码范围: 65-72
• 页数: 8页
• 语种: 英文
• DOI: 10.19852/j.cnki.jtcm.20210624.001

摘要

OBJECTIVE:To investigate the potential mechanism by which Shugan Huoxue Huayu Fang(疏肝活血化瘀方,SGHXHYF)ameliorates liver fibrosis.METHODS:Liver fibrosis was induced in rats by intraperitoneal injection of carbon tetrachloride(CCl4)in peanut oil solution(40％,3 mL/kg body weight)twice a week for 8 weeks.A normal control group received the same volume of peanut oil alone.During weeks 5-8,the CCI4-injected rat groups were administered saline(vehicle control),colchicine(0.1 mg/mL,1 mg/kg,positive control),or SGHXHYF(0.1 mg/mL;0.3,0.6 and 1.2 mg/kg)once daily by oral gavage.Rats were sacrificed 24 h after the last treatment.Blood samples were collected for measurement of serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(ALP),albumin(ALB),collagen Ⅰ,and collagen Ⅲ levels.Liver samples were analyzed by histopathological staining,Masson's staining of extracellular matrix proteins,and immune-ohistochemical staining of α-smooth muscle actin(α-SMA).TGF-β1/Smad protein and mRNA levels were analyzed by Western blot and quantitative reverse transcription-polymerase chain reaction analysis,respectively.In vitro experiments were also performed using rat hepatic stellate cells(HSCs).RESULTS:Compared with the control animals,CCl4-exposed rats exhibited elevated serum levels of ALT,AST,ALP,collagen Ⅰ,and collagen Ⅲ;reduced serum levels of ALB;and increased collagen deposition and α-SMA expression in liver sections,reflecting liver fibrosis.CCl4 also increased expression of TGF-β1 and the activated(phosphorylated)forms of Smad2 and Smad3 but reduced expression of the negative regulator Smad7 in the liver.Notably,concomitant administration of SGHXHYF to CCl4-exposed rats was found to significantly reverse or abolish the pro-fibrotic effects of CCl4 in the liver and reduced serum transferase levels.Analysis of HSCs in vitro confirmed that,mechanistically,SGHXHYF inhibited activation of the TGF-β1/Smad signaling pathway by downregulating phosphorylated Smad2 and Smad3 and upregulating Smad7 levels.CONCLUSION:SGHXHYF ameliorated CCl4-induced liver fibrosis by inhibiting the TGF-β1/Smad signaling pathway.These findings suggest that SGHXHYF may have clinical utility for the treatment or prevention of liver fibrosis.