Expression Of Interferon-gamma Inducible Protein-10and Its Receptor In Various Type Of Hepatitis B

Objective: Hepatitis B is an infectious diseases caused by hepatitis Bvirus (HBV), whose mainly lesions is liver inflammatory, can cause multipleorgan damage. The disease was prevalent worldwide. Pathogenesis of hepatitisB is not clear. It is considered that immuno-pathological damage plays animportant role. The lesions showed a significant inflammatory features thatthe degeneration and necrosis of liver cells and inflammatorycell-infiltration.Chemokines is low molecular weight (mostly8-10KD) protein thatcontrol leukocyte migration. Chemokines binding with its receptors that canpromote development, maturation, migration, aggregation. Inflammatory cellsmediate inflammation by releasing inflammatory factors. Chemokines take aimportant part in infections, also in angiogenesis, tumor growth and metastasis,damage and repair. Interferon-gamma inducible protein-10(IP-10) belongs tothe CXC chemokine subfamily, and its only receptor is CXCR3. The immuneresponse to HBV is mainly Thl cell inflammatory response. Thl cells expressthe chemokine receptor CXCR3and CCR5. The increase of IP-10level cangather T lymphocytes which express CXCR3to the area of immunoreactionthrough the blood circulation. Interaction between IP-10and CXCR3makeTh1participate in the process of tissue damage. It is reported that, in the liver,the main source of IP-10is the liver parenchyma cells. The secretion ofchemokine by liver parenchyma cells is a way that infection site attractedeffector cells, and can remain the effect cells stay in the liver. Current studyshowed that the expression of IP-10in peripheral blood and liver tissues ofchronic hepatitis C (CHC) increase and serum IP-10level increase with thedegree of liver damage. The expression of IP-10decreased after antiviraltherapy, and the basal level of IP-10can predict therapeutic effection. Thepresent study found that IP-10take part in the chronic mechanism of hepatitis B. However, the correlation IP-10with progression or outcome of HBVinfection has no precise repoets.This study aim to explore the changes and clinical significance of IP-10and CXCR3in patients with different types of chronic hepatitis B, bydetecting the level of serum IP-10, IP-10and CXCR3in liver tissues.Methods:1Research objects: Patients in the third hospital of hebei medical universityfrom December2010to December2011were selected, including38patientswith chronic hepatitis B (mild-moderate group20cases, severe group18cases),18patients with discompensated liver cirrhosis (LC), and40patientswith chronic severe hepatitis B (CSHB). According to the pretherapy MELDscore, patients with CSHB were divided into two groups, the MELD>30group(27cases) and the MELD≤30group(13cases). The diagnosis accorded withPrevention and Treatment to Virus Hepatitis in2000and Chinses Managementof Chronic Hepatitis B in2010.15people with healthy examination wereselected as control group.2Specimen collection and preservation: took3ml whole blood of subjects atearly morning, standed at room temperature30min, then centrifuged at1500r/min for15min, drawed the supernatant, splited charging, conserved at-80℃, prepared for detection. Liver tissue removed by the liver puncture andplaced10%formalin-fixed and paraffin-embedded sections.3Liver function and renal function were detected by automatic biochemistryanalyzer. Coagulation function was determined by Automated BloodCoagulation Analayzer. HBV serological markers (HBsAg,HBsAb,HBeAg,HBeAb,HbcAb) and anti-HAV, anti-HCV, anti-HDV, anti-HEV, anti-HIVwere measured by ELISA. HBV DNA load is measured by quantitative PCR.4IP-10were recorded by ELISA.5Hematoxylin-eosin (HE) and Masson staining were used for observepathological changes and liver fibrosis. Immunohistochemistry (PowerVisionTMmethod) was used to strain IP-10and CXCR3of liver tissue.Brownish yellow coloring as a positive expression, Multifunctional pathological image analyzer is used to calculate the average surface density ofIP-10and CXCR3.6All data was analyzed by SPSS13.0for Windows software.Results:1Changes of serum IP-10levels: The levels of serum IP-10increase graduallyin CHB mild-moderate group (200.40±94.95), CHB severe group (531.52±156.58), CSHB group (717.96±173.85) compared with those in the controlgroups (78.58±40.32)(F=9.695, P<0.05). The levels of serum IP-10indecompensated liver cirrhosis group (471.10±239.13) and CHB severe groupshow no significantly difference (P<0.05).The levels of serum IP-10of MELD>30group (815.69±91.83) washigher than the MELD≤30group (669.10±185.60)(t=2.579, P <0.05).The dynamic observation: serum IP-10levels decreased as theimprovement in two CSHB ameliorated patients, and serum IP-10levelsincreased with worsening in three CSHB aggravated patients.2The expression of IP-10and CXCR3in liver tissues: IP-10was found to beselectively upregulated on the cytoplasm of the hepatocyte, bile ductulesepithelia and sinusoidal endothelium cell, and mainly accumulated in necroticarea. CXCR3was widely expressed on lymphocytes and hepatocyte. Theexpression of CXCR3increased in the portal area, necrosis area, andproliferation of fibrous area, and increased with the degree of inflammationand fibrosis.3Correlation analysis: In various types of chronic hepatitis B patients, IP-10was significantly correlated with ALT (r=0.683, P=0.008), AST(r=0.426, P=0.006), TB (r=0.383, P=0.001) and PTA(r=-0.622, P=0.001).IP-10showed significantly correlated with HBVDNA (r=0.469, P=0.005) in CHB patients.The expression of IP-10and CXCR3in liver tissues (r=0.631, P=0.005) showed signifi-cantly correlated.Conclusions:1The levels of serum IP-10increase gradually in CHB mild-moderate group, HB severe group, CSHB group and was significantly positively correlatedwith ALT, AST and TB, and negative correlation with PTA. It is suggestedthat serum IP-10level may be a indicator to predict the severity of liverdamage.2IP-10was found to be selectively upregulated on the cytoplasm of thehepatocyte, and mainly accumulated in necrotic area. CXCR3was widelyexpressed on lymphocytes and hepatocyte. The expression of CXCR3increased in the portal area, necrosis area, and proliferation of fibrous areaincreased. The expression of IP-10and CXCR3were increased with thedegree of inflammation and fibrosis. It is showed that IP-10was involved inthe immuno-pathogenesis of liver after HBV infection.3Serum IP-10level is significantly correlated with MELD in CSHB group,that may affect the prognosis.