| Objective Chemokines IP-10 is a newly discovered CXC chemokines,it comes from the activated fibroblasts,mononuclear macrophages,endothelial cells and lymphocytes and so on.It plays an important role in the development of viral hepatitis and liver immune injury.Research proves that the baseline IP-10 levels is associated with the degree of HBV infected liver inflammation,and patients with high levels of baseline IP-10 achived a better outcome of antiviral treatment.But previous studies have found that the baseline IP-10 expression was different in patients with different degree of liver inflammation and liver fibrosis,so the predict value of baseline IP-10 level in antiviral treatment was still not clear in patients with different condition.Most of the research at home and abroad was focused on serological studies,lots of clinical datas indicated that the serological indexes and histological indices of patients was not completely consistent,the real effect is given priority to the liver pathology.So the research of IP-10 changes in liver tissue of CHB patients was emparty,especially the serological and histological IP-10 dynamic changes after antiviral treatment.So we should focuse on individualized antiviral treatment for patients with different condition of CHB,deeply explore IP-10 dynamic changes in liver tissue in the process of antiviral treatment.we examined the IP-10 expression in serum,peripheral blood and liver and IP-10 dynamic changes during antiviral treatment,analyzed the relationship between the clinical features and IP-10 level.In addition,we explored the role of IP-10 in liver damage mechanism in vitro experiments,in order to provide accurate and strong help for clinical diagnosis and treatment.Methods In the people's liberation army 88 hospital and the people's liberation army 302 hospital to collect the plasma and liver biopsy tissue samples of chronic viral hepatitis b(CHB)patient in different liver inflammation and fibrosis before and after entecavir antiviral treatment,including 54 cases plasma specimens before and after enticavir treatment in patients with CHB and 35 cases liver biopsy tissue samples before and after enticavir treatment in patients with CHB,10 whole blood samples before enticavir treatment in patients with CHB,8 liver biopsy tissue samples before and after antiviral treatment in CHB.In addition,14 patients with cirrhosis and 17 healthy controls participated in the study.(1)explore the IP-10 expression level in plasma before and after enticavie treatment with ELISA kits in patients with CHB,and analyze the relationship between IP-10 expression and the degree of inflammation and fibrosis;analyze the correlation between IP-10 expression and the clinical index;(2)measure the IP-10 expression in patients with CHB in liver tissue using q PCR quantitative detection,and analyzes the difference between CHB patients and healthy controls and difference between CHB patients with liver cirrhosis;analyse the relationship between IP-10 m RNA and the degree of inflammation and fibrosis;analyse the relationgship between plasma IP-10 level and IP-10 m RNA level in the liver tissue;analysis the correlation between IP-10 m RNA level in the liver tissue and the clinical index;(3)using immunohistochemical staining method analysis the IP-10 expression difference between CHB patients with a good outcome and CHB patients with a bad outcome in lobular area and in portal area before and after enticavir treatment,and the differences of IP-10 drop level after treatment.(4)detect expression difference of mononuclear cell source IP-10 level in peripheral blood between in CHB patients and healthy controls,and the difference between CHB patients and liver cirrhosis patient,CHB patients with health control;(5)in vitro experimental to verification IP-10 has a promoting effect on liver cell apoptosis,and the inhibition effect of liver cell proliferation.Results 1.54 CHB patients were divided into G?2 group 28 cases and G>2 group 26 cases according to the liver inflammation degree,the baseline serum IP-10 level in G>2 group was obviously higher compared to G?2 group,which indicated a remarkable difference(p=0.015).54 CHB patients were divided into S1-2 group 11 cases,S3 group 21 cases,S >3 group 22 cases according to the liver fibrosis degree,serum IP-10 expression increased obviously in S3 group than S1-2 group,difference have statistical significance(p = 0.005),serum IP-10 expression increased obviously in S>3 groups than S1-2 group,the difference is statistically significant(p = 0.014).2.Baseline serum IP-10 level was positively related to liver damage index in 54 CHB patients,baseline serum IP-10 was positively correlated with baseline serum ALT(r=0.508,p<0.0001),baseline serum AST(r =0.587,p<0.0001),baseline serum HBV DNA(r=0.473,p=0.473).Baseline serum IP-10 were positively correlated with baseline serum HBs Ag(r=0.486,p=0.486),baseline serum IP-10 was positively correlated with serum HBs Ag decline after entecavir antiviral treatmen(r=0.586,p<0.0001).Moreover,serum IP-10 decline after enticavir treatment was also positively correlated with HBs Ag decline after entecavir treatment(r 0.69,p<0.0001).3.Intrahepatic baseline IP-10 m RNA level increased significantly in 35 CHB patients compared to 4 liver cirrhosis patients,difference have statistical significance(p=0.003).Intrahepatic baseline IP-10 m RNA level was significantly increases in 35 CHB patients compared to 7 healthy controls,difference has statistical significance(p= 0.002).35 CHB patients were divided into 13 cases S1-2 group and 22 cases S3 group according to the liver fibrosis degree,intrahepatic IP-10 m RNA level were significant rised in S3 group than S1-2 group,the differences statistically significant(p=0.047).While 35 CHB patients were divided into 17 cases G1-2 group and 18 cases G3-4 group according to the liver inflammation,IP-10 m RNA level was significant rised in G3-4 group than G1-2 group,the differences statistically significant(p=0.001).In addition,the serum baseline IP-10 level was positively correlated with intrahepatic baseline IP-10 m RNA levels(r=0.49,p=0.49),and the intrahepatic baseline IP-10 m RNA level was positively correlated with HBs Ag decline after entecavir treatment(r=0.34,p=0.34).4.By immunohistochemical staining analysis,IP-10 expression was significantly different before and after enticavir treatment in lobular area in 4 CHB patients with a good outcome,difference have statistical significance(p=0.001).In 4 CHB patients with a bad outcome after enticavir treatment,IP-10 expression was not obvious different before and after enticavir treatment.4 CHB patients with a good outcome after entecavir treatment has a higher IP-10 baseline level than 4 CHB patients with a bad outcome after entecavir treatment in lobule area,the differences was statistically significant(p=0.003).4 CHB patients with a good outcome after entecavir treatment has a significant IP-10 decline than4 CHB patients with a bad outcome after entecavir treatment the in lobular area,difference have statistical significance(p=0.001),4 CHB patients with a good outcome after entecavir treatment have a significant IP-10 decline than 4 CHB patients with a bad outcome after entecavir treatment the in portal area,difference have statistical significance(p=0.043).5.Analysis mononuclear cells source IP-10 in peripheral blood by flow cytometry staining,IP-10 express in peripheral blood has a significant rise in 10 CHB patients than 10 patients with liver cirrhosis,the difference was statistically significant(p<0.001),IP-10 expression has a significant rise in 10 CHB patients than 10 healthy controls in peripheral blood,the difference was statistically significant(p<0.001).IP-10 expression has a significant rise in 10 liver cirrhosis patients than 10 healthy controls in peripheral blood,the difference was statistically significant(p< 0.001).6.In vitro experiment,Hep G2 72 h proliferation rate was 73.5% in control group,100 ng/ml IP-10 stimulating Hep G2 72 h,Hep G2 proliferation rate was 72.3%,10 ng/ml IP-10 stimulating Hep G2 72 h,Hep G2 proliferation rate was 51.3%,liver cell proliferation was significantly suppressed.In control group Hep G2 apoptosis rate in 6h was 0.365%,10ng/ml IP-10 stimulus Hep G2 6h,Hep G2 apoptosis rate was 0.35%,100 ng/ml IP-10 stimulus Hep G2 6h,Hep G2 apoptosis was 0.38%,1 ng/ml IP-10 stimulus Hep G2 6h,Hep G2 apoptosis rate was 0.595%,liver cell apoptsis was significantly increased.Conclusions 1.Baseline serum IP-10 level was positively correlated liver inflammation and fibrosis degree in CHB patients,.2.Baseline serum IP-10 level in CHB patients was positively correlated with baseline serum ALT,AST,HBV DNA,HBs Ag,.3.Baseline IP-10 m RNA level was higher in CHB patients.Intrahepatic baseline IP-10 m RNA level was positively correlated with liver inflammation and fibrosis degree in CHB patients.Baseline intrahepatic IP-10 m RNA level was positively correlated with HBs Ag decline after entecavir treatment.Intrahepatic baseline IP-10 m RNA level was positively correlated with baseline serum IP-10 level in CHB patients,4.Baseline IP-10 level was significantly higher in CHB patients with a good outcome than CHB patients with a bad outcome in lobule area.IP-10 expression in CHB patients with a good outcome has a significantly decreased after entecavir treatment.5.Mononuclear cell source IP-10 level in peripheral blood in CHB patients was significantly increased.6.IP-10 can promote liver cell apoptosis,inhibiting liver cell proliferation. |
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