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.

LIU Lei;GUO Hanbin;SHAO Cuiping;WANG Lin;XU Youqing;ZHOU Yiming

Department of Gastroenterology,Beijing Tiantan Hospital,Capital Medical University,Beijing 100070,China;Department of Hepatology,the seventh medical center of the People's Liberation Army General Hospital,Beijing 100700,China

中医杂志（英文版）

[1]LIU Lei;GUO Hanbin;SHAO Cuiping;WANG Lin;XU Youqing;ZHOU Yiming-.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)[J].中医杂志（英文版）,2022(01):65-72

SGHXHYF,ohistochemical

Shugan,Huoxue,Huayu,Fang,疏肝活血,活血化瘀方,attenuates,carbon,tetrachloride,induced,hepatic,fibrosis,rats,by,inhibiting,transforming,growth,signaling,OBJECTIVE,To,investigate,potential,mechanism,which,ameliorates,liver,METHODS,Liver,was,intraperitoneal,injection,CCl4,peanut,oil,solution,body,weight,twice,weeks,normal,control,received,same,volume,alone,During,CCI4,injected,groups,were,administered,saline,vehicle,colchicine,positive,once,daily,oral,gavage,Rats,sacrificed,after,last,treatment,Blood,samples,collected,measurement,serum,alanine,aminotransferase,ALT,aspartate,AST,alkaline,phosphatase,ALP,albumin,ALB,collagen,levels,analyzed,histopathological,staining,Masson,extracellular,matrix,proteins,immune,smooth,muscle,actin,SMA,TGF,blot,quantitative,reverse,transcription,polymerase,chain,reaction,analysis,respectively,In,vitro,experiments,also,performed,using,stellate,cells,HSCs,RESULTS,Compared,animals,exposed,exhibited,elevated,reduced,increased,deposition,expression,sections,reflecting,activated,phosphorylated,forms,Smad2,Smad3,but,negative,regulator,Smad7,Notably,concomitant,administration,found,significantly,abolish,fibrotic,effects,Analysis,confirmed,that,mechanistically,inhibited,activation,pathway,downregulating,upregulating,CONCLUSION,ameliorated,These,findings,suggest,may,have,clinical,utility,prevention

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