Study Of Effect Of Activin A On Alcoholic Cirrhosis

Liver fiberosis is the main expression of pathology, which advance stage is cirrhosis. It is known to all that cirrhosis is one of lethal disease which lead to death. The mechanism of liver fiberosis is quite complecate that comes from extra cell matrix result by liver tissue damage and inflammation. The cirrhosis is comes form the liver fiberosis which leaded by chronic liver damage . Alcoholic liver diseases (ALD) is one of normal and defencive disease, and the reason is definitly clear, which resulted in long period overage drink. ALD is the summar of alcoholic fatty liver( AFL), alcoholic hepatitis( AH), alcoholic fibrosis(AF), alcoholic cirrhosis(AC). The incidence of world wild ALD and the followed ecnomical burden is kind of increasing trend, which lead to the wildly concern of alcoholic cirrhosis defence research. ALD which leaded by long period overage drink has became the second cause of liver disease. Morden medical science has done the research of ALD for decades, from the metabolize method of alcohol,the formation mechanism,diagonse means and defence of ALD. We has done the investigate of gene expression and control of ALD followed the application of molecule biology technolgy. Although the pathology form change of ALD has became quite clear, the mechanism of pathology damage is still a mysery.Activin is a member of the transforming growth factor-beta (TGF-β) superfamily as growth and differentiation factors, not only can regulate secretion of follicle-stimulating hormone (FSH) and follistatin (FS) from anterior pituitary, but also can serve as an important regulator in the process of embryo formation, nerve differentiation, hematogenous cells proliferation, tissue repair and inflammation recovery. The new research has found that Activin A could lead to liver fiberosis derectly, and liver fibrosis induce factor TGF-βis also partly depend on the induce function of endocrism Activin A. As a result,we first diagnose the relationship detween the level of chronical liver disease and ALD patients serum Activin A and the liver pathology change, to investigate the function of Activin A in the formation of liver fibrosis. Though the further investigate of the expression of liver tissue Activin A,Follistatin(FS)and activin IIA receptor,to discuse the funcion of Activin A in the formation of ALD;The experiment also though the ALD mouse model,further investigate the funcion of Activin A in the formation of ALD。The research will reveal the funcion of Activin A in the chronicl liver disease and the formation of ALD,to provide the new evidence for the diagnosis and cure of liver fibrosis.We carry out the research as followed:1,the veration of the level of Activin A in HBV pateints serum We use ELISA to mensurate the Activin A,hyaloplasm (HA),typeⅣcollagen (CIV),laminin (LN) content of 118 chronic HBV pateints ( longitudinal 35, sever 30, serious 28,cirrhosis 25) and 20 control group serum, and analyse its relativity. At the same time we use HITACHI 7600 biochemistry analyse apparatus to determine liver function (TBil,ALT,GLB) and to analyse its relativity. Liver biospy and HE stain has been carried on 35 patients. According to pathology grade and stage, to analyse the variation of Activin A level. The result shows that Activin A level in chronic HBV at light,middle and heavy degreee and liver cirrhosis(LC) patients increase on by on,much higher than control group(P<0.01). Activin A is in obviously positive correlation with HA,IVC and LN(P<0.01或P<0.05). Activin A is in positive correlation with Hepati functional parameter(TBil,ALT,GLB) (P<0.01).Activin A level variation is according to the pathology grade and stage of hepatic tissue.2,the relationship of Activin A and other factor which resulte in liver cronic injuryWe adopt ELISA to determine Activin A level in serum in the people include with 30 pregnant women who suffer intrahepatic cholestasis of pregnancy (ICP),25 atutoimmune liver diseases(AILD),50 nonalcoholic fatty liver disease, (NAFLD) and 20 healthy adults(control), we use Hitachi 7600 biochemistry autoanalyser to determine hepatic function parameter(ALT,AST,TBA),and analyse the correlation between the Activin A and hepatic functional parameter. The activin A level in ICP,AILD,NAFLD patients'serum is obviously higher than control group, statistics analysis shows significant deviation(P<0.01). The activin A level in ICP,AILD,NAFLD patients'serum is in positive correlation with Hepatic functional parameter (ALT,AST,TBA)(P<0.01;P<0.05), and is in positive correlation with the Hepatic functional lesion degree,by turns,C level > B level > A level.Activin level among the groups of NAFLD,AILD,ICP has no significant deviation.3,To determine the Activin A level in ALD patients serumand and the Activin A mRNA in peripheral blood circulation.Adopt ELISA to examine the activinA level in blood serum in 30 alcoholic cirrhosis and 20 healthy adults.Meanwhile we adopt Hitachi 7600 biochemistry autoanalyser to determine hepatic function parameter(ALT,AST,GGT).And we compare the dependablity between Activin A and NO,Activin A and ALT,Activin A and AST,Activin A and GGT. NO's determination apply chemical method.We apply fluorescent quantitation RT-PCR to investigate the variation of paeriod blood circulation FS mRNA level in the ALD patients and the control group.The result show that the Activin A level in AC patients serum is obviously higher than in control group, statistics analysis shows significant deviation(P<0.01).Activin A level in AC patients serum is in positive correlation with NO and Hepatic functional parameter(ALT,AST,GGT).Activin A level in AC patients paeriod blood circulation is obviously higher than in control group, statistics analysis shows significant deviation(P<0.01).4,Immunohistochemistry staining examine the expression of Activin A,FS and ActRⅡA in the tissue of alcoholic cirrhosisAdopting rabbit's Activin A and ActRⅡA polyclonal antibody,mice anti-FS monoclonal antibody to do immunohistochemistry staining to examine the expression of Activin A,FS and ActRⅡin the liver tissue of Aalcoholic cirrhosis,the result indicate that the masc grains in immunohistochemistry staining of Activin A and ActRⅡA's antibody distribute in the area around hepatic tissue header which suffer chronic Hepatic injury,while it express in low level in the normal hepatic tissue.There's no obvious contrast between the FS expresion level in the hepatic tissue in alcoholic cirrhosis and in normal hepatic tissue.Further indicate that the high expression of Activin A and ActRⅡA may be in the relationship with the formation and development of the hepatic cirrhosis.5,The expression of Activin A,FS and ActRⅡA in the tissue of experimental alcohol liver damnification mice modelThe experiment adopt 56% alcohol to irrigate stomache to preparate the experimental alcohol liver damnification mice model (alchol 5g/kg weight), The result reveals,normal liver express Activin A mRNA,it express more during the course of liver damnification,its peak value is at the third day after irrigate stomache with alcohol,paralleled with the variety trend of the ALT,AST level. Immunohistochemistry pigmentation result indicate that normal mice liver express Activin A,FS and ActRⅡA in low level,mainly locate in the liver cells in the vena portae hepatis area in acini hepatis , during the course of liver damnification, Activin A and ActRⅡA expression level increase obviously,furthermore around the area where the liver cell's have been damaged,they express most obviously;while FS increase not quite obviously. Excecute these mice 1,3.,5 and 7 days after irrigate stomache ,adopt half ration RT-PCR and immunohistochemistry to examine Activin A mRNA and Activin A,FS and ActRⅡA protein expression, examine mice's ALT,AST contemporarily.From the exprients of above we could concluded:1. The test of Serum Activin A can be the basis to judge the level of the broken in liver cell and the fibersis of liver.2. Activin A is a cell factor with multiple function,it has a wide range of biological function ,it's main function in liver is restraining the liver cell's hyperplasia,guiding cell's death\promote ECM's secrete.Activin A lead an important role in the development of the process of forming liver's relive and liver fibrosis.The research find that the level of Activin A both in the serum and liver in patient of alcoholic liver cirrhosis crease,further means that the excessive expression of Activin A may be the cause of forming alcoholic fibrosis and liver cirrhosis3.The combination of FS and Activin A can block the biological effection of Activin A, Activin A-FS system leads an important role of adjusting multiple physiological balance,if every aspect express innormaly,the physiological balance will be broken and illness will caused.Our research find in the body of the patient of alcoholic fibrosis,expression of Activin A increases,while the expression of FS not change obviously,which means,the loss of balance of Activin A-FS system may be the causion of the development of alcoholic fibrosis and liver cirrhosis4.This research make use of the modal mice who have the liver broken due to alcohol in experiment to further improve that the enliven of Activin A-ActRIIA system while the loss of the balance of Activin A-FS system may be the cause of forming fibrosis and liver cirrhosis.The above research point out ,if we can increase the secrete of interpreme FS or replenish the input of extrapreme FS,in both ways can we achieve the goal to cure the liver fibersis.The research can not only help people to understand the reason of forming acoholic fiber cirrhosis further,but also provide new evidents and trains of thought.