Experiments On Oxymatrine Preventing And Curing Liver Fibrosis

Liver fibrosis is a kind of pathological process of abnormal deposition ofconnective tissue in liver resulting from the unbalance of production anddegradation of collagen protein and other extracellular matrix (ECM), whenhepatocytes necrosed or stimulated by inflammation. Light liver fibrosis isdefined as fibrosis, and severe liver fibrosis will further result in thereconstruction of hepatic lobules and the formation of pseudo lobules andnodes, which are defined as hepatocirrhosis.Different pathogenesis, such as virus, alcohol, parasite and so on lead todifferent liver chronic damage proceses, but it is currently believed that thecommon pathway of liver fibrosis induced by different pathogenesis is thatactivated hepatic stellate cells (HSC) transform to myofibroblasts (MFB),ECM components are produced excessively and degraded relativelyinsufficiently, as well as a large quantity of collagen and other ECMdeposited in liver. The process of the formation of liver fibrosis is notinvariable, but develops constantly. With regards to this process, it is currentlyaccepted that damaging factors, such as viral hepatitis, schistosomiasis,chemical toxic substance(alcohol, CCl4 and so on)lead to the damage andnecrosis of hepatocytes. Besides, Kuppffer cell(KC) are also activated toexcrete many cytokines like transforming growth factor-β(TGF-β),platelet-derived growth factor(PDGF), interleukin-1(IL-1) and tumor growthfactor(TNF). The cytokines excreted by KC, platelets, the sinusoidsendothelial cells and hepatocytes as well as some chemokines can activateresting hepatic stellate cells(HSCs). Which in reverse will be activated andtransform to myofibroblast (MFB) (phase Ⅰor early inflammatory responsephase),then proliferate(phase Ⅱor inflammatory response phase) in responseto self-secretion and bypass-secretion. MFBs further produce and excrete alarge amount of ECM (phase Ⅲor later inflammatory response phase)stimulated by the self-secretion of interferon-α(IFN-α), TGF-β, fibroblastgrowth factor (FGF). In addition, many evidences indicate that HSCscontribute to the decrease of degradation of ECM, which result in theexcessive deposition of ECM in liver. And the interaction of vascularendothelial growth factor (VEGF), PDGF and basic fibroblast growth factor(bFGF) produced by hepatocytes can make phenotype changing ofendotheliocytes, which leads to liver sinus capillarization, collagen depositionat clearance under endotheliocytes, continuous basal membrane formationbetween liver sinus and hepatocytes, Disse cavity becoming narrow andblocking oxygen and nutriment exchange between liver sinus and hepatocytes,All of these further induce hepatocytes activity declining, artery pressureincreasing, fibrous tissues proliferation and formation of liver fibrosis. Withthe popular applications of molecular biology and its techniques in recentyeare, the mechanism of liver fibrosis is discussed deeply. Liver fibrosis is notinreversible, but reversible, so the study of liver fibrosis has become a hotpoint in the world. Pathology study indicates that liver fibrosis first occurs at portal area,around hepatocytes and around proliferating little bile duct. Liver fibrosis inportal area is that fibre extends around radially, many fibrocytes proliferate,hyperplastic collagen fibrin in portal area and central vein connects and formspace and further compart the intrinsic hepatic lobules into pseudo lobules. There are 3.6 hundred million of people in the world that are infected byhepatitis B virus, and 1.2 hundred million in China. Virus actively replicatesand liver appears inflammatory response in 50 to 70 percent of chronicinfectors, some of whom will further round into liver fibrosis, hepatocirrhosisand liver cancer, one of the main disease of human death. Liver fibrosis patientmay have no obvious symptom or behave as chronic hepatitis involvingvapidity, anorexia, abdomen, liver secret anguish and so on. The therapy ofliver fibrosis include two parts: one is aiming at pathogeny, such as antivirustherapy, antischistosomiasis therapy, abstinence wine,dispelling iron,dispelling copper and so on; the other is aiming at liver fibrosis itself, such asrestraining the activation of HSCs, suppressing the proliferation of collagen,accelerating the degradation of collagen and so on. Recently Chinesetraditional medicine has been used widely in liver fibrosis experiments andclinical studies, which provide many experiences for clinical therapy, andexperiments have proved that Chinese traditional medicine is promising inliver fibrosis therapy. Oxymatrine (OM) is a kind of alkaloid extract from Chinese traditionalmedicine bitter bean and bitter ginseng root with molecular formula of C15H24N2O2 and relative molecular weight of 264.4. Basic and clinical studieshave detected that OM has the effect of anti-inflammation, antitumor,modulation immunity, anti-hepatitis B and anti-hepatitis C. In our study, weused tetrachloride to induce lever fibrosis models in mice, and set up normalcontrast group, model group, OM prevention group and OM therapy group.Evaluating indexes includes liver function, liver fibrosis index, liver tissuepathology examine and immunochemical dying. The results of this studyindicate that OM can distinctly decrease the level of HA,LN,PC-Ⅲ,IV-CLin the serum of experimental fibrosis mice, and the effect in OM prevent groupis more obvious, and early administration of OM is better than using it later.Our results also show that HA,LN,PC-Ⅲ,IV-CL level in mice's serum ofCCl4 model group, OM prevent group and OM therapy group all markedlyhigher than normal control group, and CCl4 model group is highest, suggestingthat the increasing of these four indexes is synchronous with the extent of liverfibrosis, so examining these four indexes simultaneously is a sensitive andreliable index for judging whrther chronic liver disease patients have liverfiber tissue proliferation. Results of Liver function examination indicate that inboth OM prevent group and CCl4 model group, the level of ALT and Tbildecline and the level of protein increases (P<0.01), indicating that OM canalleviate hepatocytes denaturalization and necrosis and improve hepatocytefunction. Pathology observation finds that HSCs decreased in portal area inOM prevent group and OM therapy group and collagen deposition decreasedobviously compared with CCl4 model group, especially in OM prevent group.Using Masson dyeing, it can be observed in CCl4 model group that fiber spacebecames thick in many hepatitis compared with OM prevent group, suggestingthat OM can suppress the production and deposition of collagen. The results ofthis study show high CTGF expression in CCl4 inducing mice liver fibrosismodel, parallelling to the degree of liver fibrosis. Immunochemical dying ofCTGF approves that OM prevent group and OM therapy group obviouslydecreased(P<0.01)compared with CCl4 model group and the expression ofCTGF in OM prevent group is lower than that in OM therapy group(P<0.05),suggesting OM can prevent and cure liver fibrosis by supressing the excretionof CTGF and the effect of preventive administration is better. These results...