Correlation Between Platelet Behavior Characteristics With Liver Disease In Hepatitis B Virus (HBV) Infection

BackgroundAfter Hepatitis B virus (HBV) infection, the patients have a protean presentationincluding asymptomatic ‘‘carrier’’ status, chronic hepatitis B, Liver cirrhosis andhepatocellular cancer (HCC), aseries of liver disease. It has fatality rate, has become apublic health problem which seriously affects the health of our people. The patheogenesisof hepatitis B is not yet clear, the study clearly patheogenesis of the disease threatmentstratagies is of great sinificance, and thus to study the patheogenesis of hepatitis B hasbeen one of the hot issues.HBV long-term infection can invade the bone marrow hematopoietic function,causing the weakened of Shan nuclear-macrophage system functions, inhibit the cellproliferation of the megakaryocyte, and so that the platelet generate to reduce. Endotoxinclearance diminished capacity, resulting in platelet aggregation, in thrombosis, alsoinflammation and innate immune response and adaptive immune response play animportant role in immune regulation. The innate immune response in the activated plateletsurface expression of P-selectin can promote the activation of platelets and a variety ofimmune cell adhesion, resulting in body injury. The adaptive immune response in CD40Lis the molecular basis of its function and signal through CD40-CD40L costimulatorymolecules involved in the humoral and cellular immune responses. In recent years, clinical studies found that patients with cirrhosis existence of the portal venous system and livervascular microcirculation clotting hypersplenism caused by portal vein thrombosis andmicrocirculation in addition to the abnormalities related to blood flow and vascular local,but also may be related to the platelet abnormal activation of related. Platelets in thenormal cycle in a resting state, but platelets release granule membrane glycoproteinrearrangements and conformational changes of activated platelets by physiological orpathological stimuli.At present, the role of platelet activation and function changes in the diseasedevelopment more and more attention. The current research focus is concentrated on thechanges in platelet membrane glycoprotein receptors, such as of CD40, CD40L andp-selectin. CD40L is a type Ⅱ transmembrane protein of261amino acids, mainlyexpressed on activated platelets and T-cell surface. CD40L and its receptor CD40to formCD40-CD40L costimulatory molecule not only plays an important role in theinflammatory response, immune regulation, and biological activities. Regulatory role incoronary syndrome, cerebral embolism, the incidence of type2diabetic microangiopathymechanism plays an extremely important role. It has gradually become a new area ofresearch. P-selectin is a transmembrane protein, exist small vivo platelet α granules andvascular endothelial cell weibel-palade. Platelets or endothelial cell do not expressP-selectin in resting state, but once activated, P-selectin immediately express on thesurface of platelets and endothelial cell. Because of the gene encoding, P-selectin isknown as two variants: the transmembrane P-selectin(CD62P) and soluble P-selectin. Theformer is expressed on activated platelet surface, is the “gold standard” of plateletactivation. The latter exists in the plasma or serum, which is expressed by someendothelial damage and platelet activation in cell surface P-selectin released into the bloodformation. It can reflect the body’s overall P-selectin levels. Thus the plasma content ofthe SP-selectin can assess platelet activation and endothelial damage. P-selectin throughits lectin regin of platelet and neutrophil and monocyte adhesion, aggregation,leukocyte-mediated oxidative damage plays an important role. P-selectin can also regulateplatelet walk and expression of CD40L, and to strengthen the combination of CD40on the cell membrane, play an important role in the mediated inflammation.HBV infection in patients is often accompanied by varying degrees of abnormalblood clotting mechanism, and even bleeding tendency, not only causing liver celldamaged, but also reducing the formation of platelets and coagulation factors,consumption increased as well as abnormal activation of platelets and other factors. Inview of the past, numerous studies of chronic hepatitis B, platelet activation, confined tothe platelet count, abnormal parameters and platelet function. Of this study is intended toplatelet activation in HBV infection from immune and inflammatory response levels,observed changes in the membrane glycoprotein, to investigate the variation of diseaseprogression in HBV infection and platelet activation, preliminary clarify HBV infection inpatients with platelet-mediated liver immune injury mechanism of action.ObjectiveTo investigate the changes of platelet CD62P, CD40L, Soluble P-selectin, PAgTand APRI(AST/PLT) with the relationship of occurrence and development of chronichepatitis B disease. To clarify the characteristics of hepatitis B in peripheral blood ofpatients with platelet behavior, the experimental data and theoretical basis for earlydiagnosis and treament of hepatitis B disease worsens.1．Test the changes in chronic hepatitis B, cirrhosis and liver cancer in patients withplatelet P-selectin (CD62P) and CD40L expression, the content of plasma P-selectin andplatelet aggregation rate (PAgT) and calculate the APRI value (AST/PLT), analysis theconnection above detection factor with liver disease.2．Investigate the mechanism of abnormal platelet activation in HBV infection, and to takeearly and effective anti-platelet therapy for the liver disease to block platelet activation. Ithas important implications to preventing the aggravation of hepatitis B disease.Methods1. Choice of subjects: Chronic Hepatitis B (CHB) group of32cases,28cases of Liver Cirrhosis (LC) group,30cases of Hepatocellular Cancer (HCC). All cases are in line withthe diagnostic criteria developed by the Xi’an meeting in September2000, viral hepatitisprevention and control program, and based on clinical manifestations, laboratory tests andCT or MRI confirmed, and excluse mixed infections of other hepatitis viruses or otherautoimmune diseases. The patients enrolled had not received antiviral therapy or drugsthat affect the immune system, and2weeks of not taking drugs that affect plateletfunction.2. Immunofluorescent flow cytometry of Beckman Coulter was used to measure thepercentage of activated platelet CD62P and CD40L3. Plymouth Health instruments Co., Ltd. PRECIL LBY-NJ4, four channel plateletaggregation the detected PAgT, to understand the level of platelet function.4. Enzyme-linked immunosorbent assay in plasma P-selectin and reagents werepurchased from Wuhan Boster Co., Ltd., instrict accordance with the instruction manual tounderstand the activation of platelets in plasma.5. Hitachi7600automatic biochemical analyzer to detect liver function parameters ALTand AST; SIGMA coagulation to detect the PT; East Asia XE-2100automatic blood cellcount to detect the PLT, and calculate the ratio of APRI, AST and PLT.6. SPSS17.0software package for data analysis, measurement data were presented asmean±standard deviation (x±S) between the two groups using analysis of variance,count data were compared using the t test, P<0.05(bilateral) to have significant difference.Results1. Platelet CD40L, CD62P and plasma SP-selectin expression in CHB group, LC groupand HCC group were significantly higher than the normal control group, and withexacerbations, there have the significant difference between diseases.2. PAgT in CHB group, LC group and HCC group were significantly lower than thenormal control group, and as the disease worsens, PAgT decline more obvious, there havethe significant difference between diseases.3. APRI values in each liver disease group was significantly higher than the normal control group, and APRI values gradually increased with the severity of hepatitis Bdisease, there was significant difference between the various disease groups.4. Platelet CD40L，SP-selectin and platelet CD62P correlation analysis, found that theywere in a positive correlation.5. CHB group, LC group and HCC group patients platelet CD40L,CD62P andSP-selectin and liver function indicators, ALT and AST, and APRI correlation analysis andfound that platelet CD40L, CD62P and SP-selectin, were in a positive correlation,suggesting that platelet CD40L, CD62P and SP-selectin increased with the decline in liverfunction are closely related.Conclusions1．HBV infection, platelet levels of CD40L, CD62P significantly higher, suggesting thatplatelet activation marker levels may become an important symbol of the liver diseaseclinical stage and severity of judgment.2．With hepatitis B exacerbations in APRI (AST/PLT), the value gradually increased,suggesting that the APRI value to a certain extent reflect the degree of severity of chronichepatitis B, as the progression CHB non-invasive indirect evaluation.3．HBV infection in patients with platelets of CD40L and CD62P and plasma SP-selectinincreased with the degree of liver dysfunction was positively related to the level of plateletactivation markers that may become one of the important sign of liver disease clinicalstage and severity of judgment.