Objective:To construct a recombinant adenovirus carrying human interleukin (hIL)-10 gene and identify its expression and protection effect on rat hepatocytes during liver injury. Methods:The hIL-10 gene was amplified by PCR from template plasmid pORF-hIL10,then digested with endonuclease and inserted into pDC315 to recombine adenovirus Ad5-hIL10. Ad5-hIL10 was used to infect rat hepatocytes,and the hIL-10 expression was identified by immunology method. Rat hepatocyte injury model was established,and protection effect of the Ad5-hIL10 was investigated both in hepatocytes in vitro and rat models in vivo. Results:The recombinant adenovirus Ad5-hIL10 was recombined successfully. After infected with Ad5-hIL10,the expression level of hIL-10 in hepatocytes was increased with viral MOIs. The cell viability of virus-infected hepatocytes increased along with viral MOIs under carbon tetrachloride induction,being 92% when MOI=50. In rat model,compared with the control group and blank virus group,the hepatocyte hydropic degeneration and stroma inflammation in Ad5-hIL10-treated group were alleviated,and the liver function was improved(P < 0.05). Conclusion:The recombinant adenovirus Ad5-hIL10 has been successfully constructed,which was demonstrated to have activity of improving rat hepatocyte viability and protecting liver function.