| Alcoholic liver disease(ALD) includes mild alcoholic liver disease, alcoholic liver Fatties(ALF),alcoholic hepatitis,alcoholic fibrosis(AF),alcoholic liver cirrhosis(AC). Traditional Chinese Medical (TCM) therapy has potential superiority and distinct characteristics in the treatment of alcoholic liver cirrhosis. By means of the methods and doctrines of Clinical Epidemiological Research, we aim at the investigation of the characteristics of traditional Chinese medical syndrome of alcoholic liver cirrhosis, attempt to explore a better way for the treatment of AC. Alcoholic fibrosis is the most phase on the way to alcoholic liver cirrhosis.As its reversibility, Alcoholic fibrosis has been recognized as one of the most exciting domains in the studies on the forming mechanisms of alcoholic liver fibrosis. The forming mechanisms of Alcoholic liver fibrosis base on the activation of hepatic stellate cell, and is the reversible dynamic pathologic process of the increased synthesis and the relatively decreased degradation of ECM. On the grounds of techniques of Cytobiology and Molecular Biology, we aim at the research in the cellular and molecular mechanisms of Radix Paeoniae Rubra Distilled Substances compound anti-AF .The paper is composed of three parts :the theoretical study ,the clinical research and the experimental study.In the theoretical part,I summarized the cognition of the cause and mechanisms of alcoholic liver disease (fibrosis / cirrhosis) by the angle of traditional Chinese medicine; summarized succeedly the development of the ALD treatment by the way of Chinese traditional medical compounds and effective components,and of the clinical epidemiology in the inland studies on ALD. On the other hand, expounded systemically the pathogenesis of alcoholic liver disease (fibrosis / cirrhosis)by the angle of western medicine.In the clinical investigation, our group studied clinical cases in multi-center prospective method. In virtue of factor Analysis, we explored the characteristics of traditional Chinese medical syndrome of alcoholic liver cirrhosis, and the forming characteristics of alcoholic liver cirrhosis in inland.In the experimental research , cellar culture was mainly taken, rat hepatic stellate cells(HSC-T6) were regarded as the study object, acetaldehyde was used as the stimulating factor, Radix Paeoniae Rubra Distilled Substances compound was the experimental drug. Through cells cultivation, we studied the inhibiting proliferation effects and the apoptosis-promoting effects of dog serum after being fed with Radix Paeoniae Rubra Distilled Substances on hepatic stellate cell (HSC-T6)builded。Through analyze the protein levels of Bcl-2 , Bax , Caspase3 in HSC-T6,we find that exposure Radix Paeoniae Rubra medicated serums can change the expression of apoptosis -related genes , partly opened out the molecular mechanisms of Radix Paeoniae Rubra Distilled Substances compound anti-AF .Result and Conclusions(1) clinical investigationExperiment one: Male was the major case among the patients of alcoholic liver cirrhosis, the mean age of AC patients was approximate to 50 years old. The incidence of AC was different with the kinds of profession, the most was businessman(38%), the second was officers(32%); The prevalence rate of AC was relatively higher when the drinkers consumed the equivalent of 160g per day with a mean duration of 20 years, which was approximate to a total alcoholic consumption of 1500kg. With increase of alcoholic consumption, The incidence of the serum levels ofγ-GT,AST/ALT,ALP,TBIL,ALB,PTA rosed.Experiment two: Among the subjects, the characteristics of TCM syndrome of AC mainly manifested excess syndrome with the common manifestations of deficiency and excess; The visceral position was mainly in liver,gallbladder,kidney and spleen; The result suggested that stagnation of qi, blood stasis, damp heat, with deficiency of yin genuine of liver and kidney and with deficiency of yang genuine of spleen and kidney were the interpretation of the causes of AC. The total alcoholic consumption,dringking time were important facts,which effected on the distribution of the characteristics of TCM syndrome of AC, so was the total alcoholic consumption,dringking time when the patients had a alcohol abuse family.γ-GT,AST/ALT,ALB,PTA,Child-pugh had mostly significant effect on the distribution of the characteristics of TCM syndrome,(2)Experiment research:1. Experiment one: acetaldehyde (87.5um) in vitro cell model stimulating hepatic stellate cell- T6 (HSC-T6) proliferation was made.2. Experiment two: We adopted HSC-T6 builded model by acetaldehyde, for study in vitro. We took the serum of a dog before and at different points of time after feeding the dog with Radix Paeoniae Rubra Distilled Substances for experiment one time.The HSC-T6 cells were cultured in mediums with different concentrations of the medicated serum at different points of time for 72 hours. The results showed that HSC-T6 inhibition rate is the highest at the containing 2% 2-hour and 3 - hour medicated serum.3. Experiment three: By means of flow cytometry, we examined the apoptosis-promoting effects of the serums on HSC-T6.. The medicated serums has significant effects of promoting HSC-T6 apoptosis at these 2h,3h points of time.Compared with non-medicated serums under the same conditions, the proportion of apoptosis had a significant increase. HSC-T6 cell's proportion was increased in G0/G1 period and decreased in the S period.4. Experiment four: we took the serum at 2-hour point after feeding as the medicated serum for experiment. By immunohistochemistry method, we analyze the protein levels of Bcl-2 , Bax , Caspase-3 in HSC-T6 . The results showed that Compared with common HSC-T6, builded model HSC-T6 increases of Bcl-2 proteins were observed , while decrease of Bcl-2 protein levels was obtained . Compared with model, HSC-T6 functed medicated serums under the same conditions, decrease of Bcl-2 proteins were observed , while increases of Bcl-2 protein levels was obtained .Conclusions: alcoholic liver cirrhosis regularly occurs among the drinkers of male, 50 years old,officer or businessmen, when the drinkers consumed the equivalent of 160g perday .with a mean duration of 20 years, which was approximate to a total alcoholic consumption of 1500kg.The characteristics of TCM syndrome of alcoholic liver cirrhosis mainly manifested deficiency and excess syndrome; The visceral position was mainly in liver,gallbladder,kidney and spleen; The result suggested that stagnation of qi, blood stasis, damp heat, with deficiency of yin genuine of liver and kidney and with deficiency of yang genuine of spleen and kidney were the interpretation of the causes of alcoholic liver cirrhosis. Through cells cultivation, we studied the inhibiting proliferation effects and the apoptosis-promoting effects of dog serum after being fed with Radix Paeoniae Rubra Distilled Substances on HSC-T6 builded model by acetaldehyde in vitro.We find the function of Radix Paeoniae Rubra Distilled Substances might be one of the mechanisms to counteract against alcoholic liver fibrosis.
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