| Objective:Alcoholic liver disease is characterized by the liver damage induced by substantive and chronical alcohol ingestion.The incidence of alcoholic liver disease is increasing in China during the recent years,which results in its second position just under virus hepatitis in liver diseases.Hereto,there is no the specific remedy for alcoholic liver disease yet at home and abroad . By contraries,owing to the relatively little side effects,Chinese traditional medicines have the broad future of application and development.Professor Li Dian gui considers that the pathogenesis of ALD is that the poison of alcoholic which full of damp-heat injures stomach, then stomach fails to proper downwards.Turbid pathogens blocked in the body , which is fighting with the poison of alcoholic, become"zhuo du"."zhuodu"which can't be removed easily is blockaged in middle warmer and staying below the costal region."zhuodu"is the key word of ALD. Professor Li Dian gui studied out Huazhuo Jiedu Decoction (HZJDD)that based on"zhuodu"is blockaged in middle warmer.HZJDD have eleven herbs,such as gegen,tianjihuang,heye,yinchen,danshen and so on.HZJDD's have many functions.Such as dissolving turbid,detoxifying, cleaning heat,excreting damp,treating the liver and stomach simultaneously.Our experimental study plan to investigate curative effect and operation mechanism of HZJDD.All these can not only provide certain curative effect formula of TCM to treat ALD, but also provide clinical and experimental basis for the theory of"zhuodu".The objective of our study is to observe the effects in pathology,serology and immunohistochemistry of huazhuojiedu method for the rats'alcoholic liver disease by animal experiment and to study the function and mechanisms of the huazuojiadu method.Through the experiment,we try to approve the feasibility and practicability of huazuojiadu therapy to treat ALD.Methods:60 male SD rats,190-210 weight,were divided into 5 groups randomly:A group serves as normal group,B group serves as model group,C group serves as Diammonium Glycyrrhizinate(Grz) group,D group serves as HZJDD low dose group,E group serves as HDJDD high dose group.Except normal group,the rest groups were given mixed liquid including Erguotou white wine and corn oil to the preparation of ALD rats. Eight weeks later, the four groups including B,C,D,E were given 0.9% Nacl i.p.,Diammonium Glycyrrhizinate,HDJDD low dose,HDJDD high dose respectively to carry on control study.We should observe rats'appearances,expressions,actions through the whole experimental process.All rats were killed after 12 weeks.Rats serum was taking using for liver function,hepatofibrosis,CAT and IL-8.Hepatic tissue is taking using for HE staining and Cox-2 immunohistochemistry.All data were analyzed with SPSS 11.5 software.Inter-group test of significance were analyzed using one-way ANOVA.Pairwise comparison were analyzed using LSD.Results:1 The ecumenic states of ratsNormal group shows slicking hair,vigorous,free activity, also diet and excrement are normal.Model group shows slow crawl,limp extremities after given the mixed liquid every morni- ng.The hair of model group is less slicker,dispirited activity and irritated easily.These rats weight decline subsequently,and some rats'excrement becomes watery.After treatment,theC,D,E group rats'resist strengthen day by day.The high dose group have the biggest resist,and Diammonium Glycyrrhizinate group have the smallest.2 Hepatic histopathology2.1 Gross observationsThe normal group livers were henna and bright.Liver's envelope are smooth and flexible.The model group livers were buff ,rough,dull and obviously large.Touching the liver,we feel harder than the normal.The sections were greasy.Diammonium Glycyrrhizinate group and Chinese traditional medicine group have no significant feature of the visual study.2.2 HE StainingNormal group:Hepatic tissue and structure were inte- grated.Hepatic lobule structure was clear.The hepatocytes arranged in radiation orderly by the center of the central veins. cell nucleus were big and round,which located in the cell's center.Model group:The primary change of this group was fatty degeneration.Hepatic cells intumesce,especially the central vein- s district.Fat droplets which differ in size could be found in the intracytoplasm.Lymphocyte and plasma cell infiltrated in the portal area.Aslo we may found Mallory bodies once in a while.Diammonium Glycyrrhizinate group:Fatty degeneration mitigated evidently.Hepatic cells intumesced slightly.Smaller fat droplets which surrounded cell nucleus could be found in some cells.A small quantity of Lymphocyte and plasma cell infiltrated in the portal area by chance.HDJDD low dose group:Fatty degeneration mitigated evidently.We can found tiny fat vacuolus and cell acidophilia change in the liver cells.Sporadic inflammation and periangiitis can be found between Hepatic lobules.HDJDD high dose group:Fatty degeneration has vanished on the whole.Small Vacuolus degeneration and inflammation in a fraction of intracytoplasm could ba found under high power lens.3 The vary of serological index in liver in ALD rats3.1 Liver function parametersCompared with normal group,AST,ALT in rats'serum of model group is higher significantly(P<0.01).Compared with model group,AST,ALT in rats'serum of the three treated groups are lower significantly(P<0.05).AST,ALT in rats'serum of the HZJDD high dose group are lower than Grz group and HDJDD low dose group(P<0.05).3.2 The active of CATCompared with normal group,CAT in rats'serum of model group is higher significantly(P<0.01).Compared with model group,CAT in rats'serum of the three treated groups is lower significantly(P<0.05).Hereof,AST,ALT in rats'serum of the HZJDD high dose group are lower than Grz group and HDJDD low dose group significantly(P<0.05).3.3 The index changes of hepatic fibrosisCompared with normal group,HA,Pâ…¢P in rats'serum of model group is higher significantly(P<0.01).Compared with model group ,HA,Pâ…¢P in rats'serum of the three treated groups is lower significantly(P<0.05).Hereof, HA,Pâ…¢P in rats'serum of the HZJDD high dose group are lower than Grz group and HDJDD low dose group significantly(P<0.05).3.4 The active change of IL-8Compared with normal group, IL-8 in rats'serum of model group is higher significantly(P<0.01).Compared with model group,IL-8 in rats'serum of the three treated groups is lower significantly(P<0.05).Hereof,IL-8 in rats'serum of the HZJDD high dose group are lower than Grz group and HDJDD low dose group significantly(P<0.05).4 The immunohistochemistry(SP)of COX-2 in liver tissue:There was no positive expression of COX-2 in hepatic tissue of normal rats.In model group,there was positive expression of COX-2 which color was brown distributed in the rats'hepatocytes cytoplasm.The intensity of expression of COX-2 had obviously increased(P<0.01).Compared with model group,the COX-2 expression of the three treated groups is lower significantly(P<0.05).Hereof,the COX-2 expression of the HZJDD high dose group are lower than Grz group and HDJDD low dose group significantly(P<0.05).Conclusion:1 The method with the mixed liquid including Erguotou white wine and corn oil to intragastric administration which duplicates the alcoholic liver disease model is successful,and the model is steady ,simple and economical.2 The Huazhuo Jiedu Decoction that is created according to the huazhuojiedu method can reverse fatty degeneration and hepatic fibrosis,protect the liver function, raise anti-oxidation capability and lessen inflammatory reaction.So we can say that it operates in preventing and treating alcoholic liver disease.3 It is the feasible and practical to treat ALD by huazhuo- jiedu method.It has provided new ideas and methods for the prevetion and treatment of alcoholic liver disease. |
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