The relationship among HBV cccDNA,HBV total DNA in liver tissue and serum HBV DNA and their clinic - pathological significances in patients with chronic hepatitis B

Objective To investigate the relationship among covalent closed circular DNA(cccDNA),HBV total DNA(tDNA) and serum HBV DNA and their clinic-pathological significances in patients with chronic hepatitis B.Methods Seventy eight patients with chronic hepatitis B were enrolled in this study.The[3- globin DNA,HBV cccDNA and HBV tDNA in liver tissue were detected by real-time PCR.The content of single hepatocyte HBV cccDNA and HBV tDNA(copies/ cell) equals to HBV cccDNA(measured value)/(3-globin DNA measured value(copies),and HBV tDNA measured value (copies)/β-globin DNA measured value(copies).The content of intrahepatic HBV cccDNA and HBV tDNA was recorded as logto copies/10~6 cell.Real-time PCR and ELISA were performed to determine serum HBV DNA and HBV markers respectively. The expression of HBsAg and HBcAg in hepatocytes was detected by immunohistochemical method.SPSS 13.0 software was adopted for statistical analysis.Results(1) There was a positive correlation between the hepatocyte HBV cccDNA and HBV tDNA(r = 0.696,P0.001),Similar positive correlation was found between hepatocyte HBV cccDNA and serum HBV DNA(r= 0.304,P0.01),.There was a positive correlation between hepatocyte HBV tDNA and serum HBV DNA(r= 0.341,P0.01) as well.(2) Serum HBV DNA in patients with intrahepatic HBcAg(+) was higher than that in patients with intrahepatic HBcAg(- ),and there was a significant difference(P0.05).There was no significant difference between hepatocyte HBV cccDNA and HBV tDNA in patients with intrahepatic HBcAg(+) and intrahepatic HBcAg (- )with P0.05 respectively.(3) Serum HBV DNA with intrahepatic HBsAg(+) was higher than that in patients with intrahepatic HBsAg(-),and there was a significant difference(P0.05).There was a positive correlation between hepatocyte HBV cccDNA and HBV tDNA in patients with intrahepatic HBsAg(+) and intrahepatic HBsAg(- ) with P0.05 respectively.(4) Serum HBV DNA in patients with HBeAg(+ )/ anti-HBe(- ) was higher than that in patients with HBeAg(- )/anti-HBe(+).and there was a significant difference(P0.05).There was no significant difference between hepatocyte HBV cccDNA and HBV tDNA in patients with HBeAg(+ )/anti-HBe(- ) and HBeAg(- )/anti-HBe(+ ) with P0.05 respectively.(5) There was no significant correlation between hepatocyte HBV cccDNA,HBV tDNA and serum HBV DNA and the levels of liver inflammation activity and fibrosis(P0.05).Conclusion(1) Serum HBV DNA cannot fully reflect the content of hepatocyte HBV cccDNA and HBV tDNA.Even when serum HBV DNA level was lower than 500 copies/mL,hepatocyte HBV cccDNA could still be detected with the maximum difference of 10 times of the contents. (2) Serum HBV DNA in patients with intrahepatic HBcAg(+) or HBsAg(+) was both higher than that in patients with intrahepatic HBcAg(- ) or HBsAg(- ),and there was no significant difference between their hepatocyte HBV cccDNA and HBV tDNA.(3) Serum HBV DNA in patients with HBeAg(+)/anti- HBe(- ) was higher than that in patients with HBeAg(- )/anti-HBe(+),and there was no significant difference between hepatocyte HBV cccDNA and HBV tDNA.(4) There was no significant correlation between hepatocyte HBV cccDNA,HBV tDNA and serum HBV DNA and the levels of liver inflammation activity and fibrosis.