Research The Effects Of HuaZhuoJieDu Prescription Based On Zhuo Du Theory Via JAK/STAT Pathway On Liver Fibrosis In Rat

Objective: As a hard medical problem with a high incidence rate, hepaticfibrosis is the common pathological development process of many chronicliver diseases, which eventually evolving into cirrhosis. Cirrhosis isirreversible while hepatic fibrosis is not. Therefore, to find out the effectiveantifibrotic drugs becomes the focus of the liver diseases study. There islacking of effective treatment in Clinical treatment while traditional Chinesemedicine （TCM）, has its unique effect with a multi-level and target treatment.My mentor professor Diangui Li first put forward and established the "Zhuodutheory". Depending on years of clinical and experimental researches, he hasfound that "Zhuo Du" is the key TCM factor in the formation of hepaticfibrosis. According to this principle, HuaZhuoJieDu prescription has achievedthe satisfactory clinical treatment effectiveness. But the mechanism of“HuaZhuoJieDu” is still unknown. At present, confirmed, hepatic stellatecell（HSC） are the main cells induced liver fibrosis. Platelet derived growthfactor（PDGF）is now known as the strongest effect on HSC mitogen. Januskinase/signal transducer and activator of transcription （JAK/STAT） signalpathway is a pathway of many cytokines, in the formation of liver fibrosis isclosely related to the process of.This experiment aims to observe the effects of“HuaZhuoJieDu” treatment on hepatic fibrosis in rats by determine itsinfluence on the expression of PDGF-BB or JAK₂/STAT₃, and then to exlorethe possible mechanism, which, thereby, providing its modernpharmacological targets and molecular biology evidence.Methods: After a one-week adaptive feeding,70Sprague Dawley malerats（body weight200±20g） were randomly divided into six groups: normalcontrol group（N）,n=10, model control group（DMN）n=20,HuaZhuoJieDu prescription high dose group（H）,n=10；HuaZhuoJieDu prescription middledose group（M）,n=10；Hua Zhuo Jie Du prescription low dose group（L）,n=10,FuFangBieJiaRuanGan tablet treatment group（BJ）,n=10, rats in each groupwere normal diet. Except the normal control group, the rats in each groupreceived intraperitoneal injection of1%DMN （10mg/kg） from everyMonday morning. On the first week of the experiment, each groupcontinuously modeling three days; From the2to6weeks continuousmodeling for two days a week, and four model rats were sacrificed in thesecond and the fourth weekend, and then observed the pathological fibrosisdegree of the liver tissue. Rat models with moderate hepatic fibrosis wereestablished till the4th week; Form5th to8th week, rats received differentdrug treatment with same volume by intragastric administration. At the end ofthe eighth week, all rats were sacrificed and fresh liver tissue was abstained,which under hematoxylin eosin staining to observe hepatic pathology; Theexpressions of PDGF-BB and JAK₂/STAT₃mRNA in liver tissue weremeasured by RT-PCR.Results:1The condition of rats of each groupThree rats of the control group were died during the experiment, and tworats of the group by accepting High doses of HuaZhuoJieDu prescription weredied. The rest of the groups were without death. The rats’ condition of controlgroup was fine, such as eating normally, moving agilely. They were havingbright fur, steady up weight and granular droppings. The rest of rats showdifferent levels of symptoms, such as eating less, sleeping more, dark fur,weight gaining slow and diarrhea.2Observation of the liver tissue’s pathologyThe rats’ livers of the control group were smooth, tenacious and brightred by macroscopic observation. Model control group rats with granularnodular liver surface, qualitative hard, dark and gloomy; Middle dose of theHuaZhuoJieDu prescription group change in party with rat liver nodules infine sand on the surface of samples, quality is a bit hard, color dark red. HE staining situation: normal control group rats hepatic lobule structure complete,liver rope order; Hepatic lobule structure damage was obviously in the modelgroup, enlargement of fibrous tissue, separated the liver lobule; Give Middledose HuaZhuoJieDu prescription of group rats change in hepatic lobulestructure c in the model group significantly reduce, fiber cords thinning.3The results of RT-PCR3.1The expression of PDGF-BB mRNA: There was a little expression ofPDGF-BB mRNA in normal control group can be detected.Compared withnormal control group, the expression of PDGF-BB was significantly increasedin model control group （P <0.01）. Compared to model control group, theexpression of PDGF-BB was significantly reduced in FuFangBieJiaRuanGantablet group, Low dose （P <0.05） and Middle dose of the HuaZhuoJieDuprescription group （P <0.01）; the expression of PDGF-BB was notsignificantly different in High doses of HuaZhuoJieDu prescription （P>0.05）.3.2The expression of JAK₂mRNA: There was a little expression of JAK₂mRNA in normal control group can be detected. Compared with normalcontrol group, the expression of JAK₂was significantly increased in modelcontrol group （P <0.01）. Compared to model control group, the expression ofJAK₂was significantly reduced in Low dose （P <0.05），Middle dose andFuFangBieJiaRuanGan tablet group（P <0.01）. The expression of JAK₂wasstay in high level, and was not significantly different in model control groupand High dose of HuaZhuoJieDu prescription （P>0.05）.3.3The expression of STAT₃mRNA: There was a little expression of STAT₃mRNA in normal control group. Compared with normal control group, theexpression of STAT₃was significantly increased in model control group （P <0.01）. Compared to model control group, the expression of STAT₃wassignificantly reduced in FuFangBieJiaRuanGan tablets group （P <0.05）.Compared to model control group, the expression of STAT₃was notsignificantly decrease in other groups （P>0.05）.Conclusions:1In the model of DMN induced rat liver fibrosis, the expression of cell factor PDGF-BB and signal transduction pathways JAK₂/STAT₃wereunregulated. This may be one of the mechanism of liver fibrosis.2HuaZhuoJieDu prescription can against rat liver fibrosis induced byDMN, improve the liver fibrosis degree and lower the expression ofPDGF-BB and JAK₂mRNA in rat liver tissue. Effect of Middle dose is moreobvious.3HuaZhuoJieDu prescription could reduce the expression of PDGF-BBand JAK₂mRNA in rat liver tissue, and accommodate JAK/STAT pathway.This pathway may be one of the role of anti liver fibrosis mechanism.