The Detection Of Gc-globulin And Cytokines IFN-γ,TNF-α,IL-4,IL-10 In The Serum Of Patients Of Liver Disease

First part:Gc globulin(vitamin D binding protein ) is mainly produced in liver. It could bind myoglobulin which releases into blood circulation after cell necrosis. It is documented that the change of Gc is obvious in liver diseases, especially in severe hepatitis .It could be an index for digonosisi and prognosis. Aim:To evaluate the concentration change of Gc in liver diseases, especially severe hepatitis, we detected concentration of free non-myoglobulin binding Gc in serum of different kinds of liver diseases in vitro with ELISA..Materials and Method :We chose 196 patients of liver diseases admitted to our hospital from Feb 2006 to Feb 2007,among which were 36 severe hepatitis(5 acute severe hepatitis,14 subacute severe hepatitis,4 acute-on-chronic severe hepatitis,13 chronic severe hepatitis),54 HBV related cirrhosis(child-pugh score:A 17,B 28,C 19),46 chronic hepatitis,15 acute hepatitis,10 drug hepatitis patients. Double sandwiches ELISA was used to detect serum Gc globulin concentration. Results: There is significant difference between severe hepatitis group and control group ,acute HBV hepatitis group ,chronic HBV hepatitis group ,HBV related cirrhosis group, drug hepatitis group separately (p<0.001). There is no significant difference among acute ,subacute ,acute-on -chronic severe hepatitis and chronic severe hepatitis group(p>0.05).Hepatocyte injury exists in various kinds of liver diseases. Then Gc globulin combinds with myoglobulin which releases into blood circulation after cell necrosis. The body clears the Gc globulin-myoglobulin complex continuously so that concentration of total and free Gc globulin decreases.Gc globulin is an acute reaction protein. Synthesis of Gc globulin in liver will increase in inflammatory or impair state in the compensation of consumed Gc.so the decrease of Gc globulin concentration in cirrhosis and acute HBV hepatitis groups are not significant than that of control group(p>0.05).and sometimes the Gc-globulin may be higher because of the increasing synthesis of Gc-globulin ,eg ,in chronic HBV hepatitis and drug group . In severe hepatitis, there is massive destruction of hepatocytes in short time which causes the fast consumption of Gc globulin combining myoglobulin. Thus Gc globulin production could not be compensated so that serum concentration decreases.Conclusion: Gc globulin is closely correlated with liver injury . Concentration of free Gc globulin in severe hepatitis patients decreased significantly. Gc globulin could be used to diagnose severe hepatitis.Second part:In the evaluation of cytokine change and its clinical significance in different liver diseases, we use double sandwich ELISA to detect serum concentration of TNF-α,IFN-γ,IL-4,IL-10. We chose 196 patients of liver diseases admitted to our hospital from Feb 2006 to Feb 2007,among which are 36 severe hepatitis,54 HBV related cirrhosis,46 chronic hepatitis,15 acute hepatitis,10 drug hepatitis patients。Results: IFN-γ,IL-4,IL-10 concentrations in acute and chronic HBV hepatitis,cirrhosis,severe hepatitis increased significantly than that of normal control group, (p<0.05),while IL-10 showed no difference between liver disease group and normal control group ,as well as among liver diseases. In drug hepatitis, only IFN-γamong the cytokines showed obvious elevation than normal group.IFN-γconcentration in cirrhosis group was significantly higher than that of chronic hepatitis B and severe hepatitis (p<0.05).IL-4 concentration in severe hepatitis was significantly higher than that of acute and chronic hepatitis B (p<0.05)The body counteracts the virus though non-specific and specific immune response. In specific immune response,T lymphocyte is the most important effector which differentiates into CTL(Tc)and T help cell.Tc clears virus by solving infected cells,but more importantly by secreting cytokines like IFN-γ,TNF-αwhich discards of virus in non-solving way.Th1 secrets cytokines like IFN-γ,TNF, mediates cell immunity. The activation of Tc is mediated by Th1. it is advantageous to clear virus when Th1 takes the superiority. Th2 cell secrets cytokines like IL-4,IL-10 ,which inhibit the function of Th1 cell. The interaction of Th1 and Th2 plays paramount role in virus infection in liver diseases. In this research , IFN-γ,TNF-α,IL-4 concentration in acute and chronic hepatitis B,cirrhosis,severe hepatitis significantly increased. Immune system is activated after virus infection establishing new balance in higher concentration of cytokines. When HBV infection exists long time,fibrous tissue is gradually formed in which cytokines plays an important part . IL-4,TNF–αstimulates fibrosis,while IFN-γ,IL-10 resist it . IL-4,TNF–α,IFN-γin chronic hepatitis B and cirrhosis rised significantly than that of normal control group which display that fibrosis and anti-fibrosis co-exist and are functioning together in the body. The network of cytokines plays an important role in pathogenesis of severe hepatitis . the concentrations of TNF–α,IFN-γwere obviously elevated than that of control group. IL-4 in severe hepatitis showed more increase than that of acute and chronic hepatitis B demonstrating its importance of immunosuppression in severe hepatitis.All in all,the cytokines of TNF–α,IFN-γ,IL-4 play an important part in HBV infection; the cytokines of TNF–α,IFN-γ,IL-4 involve in the fibrogenesis and anti-fibrogenesis in the HBV infection; proinflammatory factor of TNF–α,IFN-γand anti-inflammatory factor of IL-4 work together in the course of severe hepatitis.