Principal Pharmacodynamics Of Baogangning Granule In Hepatic Fibrosis

BackgroundHepatic fibrosis is the common pathology in the development of chronic hepaticdiseases towards liver cirrhosis. Hepatitis viruses, alcohol, drugs, poisons, blood fluke,and so on, may irritate liver long term, which then activates hepatic stellate cells(HSC) living in hepatic sinusoid to excrete excess extracellular matrix (ECM). Thepathological deposition of ECM leads to abnormal liver structure or/and function, onabroad, especially in Europe and America, alcohol is the main cause, however, in ourcountry hepatitis viruses, especially persistent infection of HBV accounts for.There are mainly two methods in the treatment of hepatic fibrosis. One is to aimdirectly at the causes by inhibiting hepatitis virus, killing blood fluke, alcoholwithdrawal, removing iron or coin, etc. The other is to suppress fibrosis itself byinhibiting the activation of HSC and proliferation of collogen, as well as promotingthe degradation of collogen.Interferon, colchicine, D-Penicillamine, lamivudine, turnout necrosis factor-a,prostaglandin, interleukin-2, and so on are the common medicine in the clinic,however, too many side-effects, unaccepetable expenses restrain the utilization inpractice. The research on hepatic fibrosis with the traditional Chinese medicine(TCM) has transmitted the focus from mono medicine onto compositus and animalmodels onto cell or molecular with determination of treatment based on pathogenesis obtained through differentiation of symptoms and signs compounding with othermethods.Baoganning granule is the outcome of supervisor Pro. Lvzhiping's years ofclinical practice. It is based on the etiopathogenisis and pathogenesis of hepaticfibrosis resulted from hepatitis, namely wet heat and toxic materials remains, liver qistagnation and blood stasis and heat blocks, which was first put forward by Pro. Lv.The recipe consists of baical skullcap root, danshen root, and so on. It was proved thatthis granule has an exact effect in reversion hepatic fibrosis after the clinical practicein TCM department of Nanfang Hospital. Funded by Military Medical Research andNSFC, we conducted researches on the mechanism of Baoganning's effects onhepatic fibrosis. This peoject researched the effects of Baoganning on rat hepaticfibrosis induced by carbon tetrachtoride and alcohol respecsivety, and alsoinvestigated the changes of transforming growth factorβ1 and tissue inhibitor ofmetalloproteinase-1 during the treatments, both of which are of great relationship inthe beginning and development of hepatic fibrosis.Now, a series of hepatic fibrosis animal models are set up. Such as, chemicalliver fibrosis, autoimmune liver fibrosis, alcoholic liver fibrosis, liver fibrosis inducedby bile duct ligation, nutrition liver fibrosis, schistosomiasis liver fibrosis and theother animal models, in order to clarify the cause and mechanism of liver fibrosis,select effective drugs, and so on.We select the rat liver fibrosis induced by carbon tetrachloride, becausepreparing this model is simple, easy, low cost and needn't long time. Further more, itsmorphological, pathophysiological changes are similar to those of human hepaticfibrosis.We choose the rat liver fibrosis induced by alcohol, which is frequently used asan experimental model to study alcohol-induced liver diseases. Preparing the model is simple, reliable, successful easily,et al.ObjectiveTo investigate the main pharmacodynamics of Baogangning granule in liverfibrosis, and analyze the mechanism in the way of the impact of Baogangning granuleon the transforming growth factorβ1 and tissue inhibitor of metalloproteinase-1.MethodsFirst, Rat liver fibrosis Model induced by carbon tetrachloride preparation: 56specific pathogen free (SPF) SD rats, half male and half female, were randomlyassigned into seven groups, with eight in each, as follows: the normal control group,the liver fibrosis model group (the model group), Baoganning high, moderate andlow-dose group, Colchicine group. Fufang Bie jia ruan gan Group (Fufang Biejiagroup).In the morning,rats in the model and the drug group received subcutaneousinjection of 40% carbon tetrachloride dissolved in peanut oil (1st 0.5ml/100g,0.3ml/100g subsequently per day); In the afternoon, drugs were given among druggroups respectively, model group was given saline (1ml/100g per day).Meanwhile,The Model and treated groups were given high-fat diet (79.5% maizena+0.5% cholesterin +20% pork fat), a normal diet to the normal group, 6 weeks in all.The end of experiment, the liver tissue were stained with hematoxylin andeosin(HE) and Mallory's trichrome according to standard procedures forhistopathological examination, the pathological changes and the collagen'sproliferation of the liver tissue were observed under the light microscope; take theupper animal serum. The activity of glutamate-pyruvate transaminase (ALT) andglutarnic oxalacetic transaminase (AST) was measured using AU5400 automaticbiochemical analyzer; the activity of the hyaluronic acid (HA),the laminin (LN),precollagen typeâ…¢(PCâ…¢) type, collagen typeâ…£(Câ…£) was evaluated according to radioimmunoassay method; the activity of transforming growth factor beta-1(TGFβ1)and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) was measured accordingto enzyme linked immunosorbent assay(ELISA).Second, Rat liver fibrosis Model induced by alcohol preparation: 56 specificpathogen free (SPF) SD rats, half male and half female, were randomly assigned intoseven groups, namely: the normal control group, the liver fibrosis model group (themodel group), Baoganning high, moderate and low-dose group, Cotchicine group.Fufang Bie jia ruan gan Group (Fufang Biejia group).In the morning,rats in the model and the drug groups were ingested drink spirits,naming "Erguotou"(56°)(0.5ml/100 per day); In the afternoon, drugs were givenamong drug groups respectively, model group was given saline (1ml/100g per day).Meanwhile, The Model and the drug groups were given innutritious diet(40% flour+20% soybean meal+20%hypo-flour+20% botan-flour+a few soybean oil, salt),besides, those rats were prohibited to eat any food except water from 20:00 everynight to the next morning before gavage, the normal group was given saline and anormal diet. 90 days in all.The end of experiment, the liver tissue were stained with hematoxylin andeosin(HE) and Mallory's trichrome according to standard procedures forhistopathological examination, the pathological changes and the collagen'sproliferation of the liver tissue were observed under the light microscope; take theupper animal serum. The activity of glutamate-pyruvate transaminase (ALT) andglutamic oxalacetic transaminase (AST) was measured using AU5400 automaticbiochemical analyzer; the activity of the hyaluronic acid (HA),the laminin (LN),precollagen typeâ…¢(PCâ…¢) type, collagen typeâ…£(Câ…£) was evaluated according toradioimmunoassay method; the activity of transforming growth factor beta-1(TGFβ1)and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) was measured according to enzyme linked immunosorbent assay(ELISA).ResultsFirst, the effect of Baoganning on CCL₄ induced hepatic fibrosis in rats.To begin with, Serum levels of ALT,AST,HA,LN,PCâ…¢,Câ…£,TGFβ1,TIMP-1 in model group were higher than those in normal group(P＜0.05); All theindexes were improved in each medicine control group(P＜0.05), especially inBaoganning high, moderate and low-dose group.In addition, the pathological pictures of the liver tissue staining by HE andMallory's trichrome showed, the model group was filled with marked fattydegeneration, obvious collagen deposition, inflammatory cells infiltration; thehepatocyte lobules were surrounded by thick bands of collagen, loss the normalarchitecture and form a pattern of micronodular cirrhosis. In Baoganning group, theliver lobule's normal structure was also destroyed, but Inflammation, fattydegeneration and collagen deposition decreased obviously.Second, the effect of Baoganning on alcohol induced hepatic fibrosis in rats.To begin with, Serum levels of ALT,AST,HA,LN,PCâ…¢,Câ…£,TGFβ1,TIMP-1 in model group were higher than those in normal group(P＜0.05); All theindexes were improved in each medicine control group ((P＜0.05), especially inBaoganning high, moderate and low-dose group.In addition, the pathological pictures of the liver tissue staining by HE andMallory's trichrome showed, the model group was filled with marked vacuolardegeneration, obvious collagen deposition, inflammatory cells infiltration; thehepatocyte lobules were surrounded by thick bands of collagen, loss the normalarchitecture and form a pattern of micronodular cirrhosis. In Baoganning group, theliver lobule's normal structure was also destroyed, but Inflammation, vacuolardegeneration and collagen deposition decreased obviously. ConclusionThe study show, First, Baoganning can resist rat liver fibrosis induced by CCL₄.Serum levels of ALT,AST,HA,LN,PCâ…¢,Câ…£,TGFβ1,TIMP-1 in modelgroup were lower than those in model group; there were small collagen depositionand inflammatory cells infiltration in Baoganning groups,while complete fibroticseptum appeared in model group. Second, Baoganning can resist rat liver fibrosisinduced by alcohol. Serum levels of ALT,AST,HA,LN,PCâ…¢,Câ…£,TGFβ1,TIMP-1 in model group were lower than those in model group; there were smallcollagen deposition and inflammatory cells infiltration in Baoganning groups,whilecomplete fibrotic septum appeared in model group.