Proteomics Analysis Of Primary Duck Hepatocytes Infected With Duck Hepatitis B Virus And Preliminary Study On Proteomics Analysis Of Primary Duck Hepatocytes Treated With DMSO

Hepatitis B virus (HBV), belonging to the Hepadnaviridae family, is a major cause of liver infection in human which is estimated that there are more than 120 million HBV carriers and 700 million HBV infection in China. Because of lack of an appropriate HBV infection system, little is known about molecular mechanisms during hepadnaviruse infection. Duck hepatitis B virus (DHBV), a member of Hepadnaviridae family, sharing similar biological features with HBV including genome organization and structural organization, is a valuable hepadavirus infection model for supporting infection with high reproducibility and efficiency and releasing the infectious viral particles. Primary duck hepatocytes (PDHs) maintained in serum-free culture medium containing 1.5% dimethyl sulfoxide (DMSO) support efficient infection of DHBV in vitro, but the mechanisms is unknown.The development of proteomic methods of two-dimension gel electrophoresis (2-DE) combined with MALDI-TOF/TOF tandem mass spectrometry (MS/MS) has enabled us to assess virus-host interactions and the changes of cellular proteins expression at a global scale, to reveal the connections between virus infection and host cellular functions. The sequenced genome of Gallus gallus (chicken) provides the possibility to identify the duck (anas platyrhynchos) proteins by MS/MS. Protemic analysis of membrane protein and cytoplasmic protein in cellular component enhance the possibility to detect the low abundance proteins in 2-DE.In the present study, we intend to utilize the DHBV-PDHs system to explore:(1) global protein expression changes during hepadnavirus infection by 2-DE; (2) global protein expression changes between PDHs treated 5% FBS and 1.5% DMSO. The differentially expressed cellular proteins were analized and clasificated according to the gene ontology criteria and pathways, and potential roles of some differentially expressed proteins in the virus-infection cells have been discussed. In the present study global changes in cellular protein expression of hepadnavirus infection were explored by 2-DE analysis, using a natural PDHs-DHBV infection system. Forty-two differentially expressed proteins in DHBV infected PDHs have been identified. Differential expression of annexin A2 and beta-actin was confirmed by Western blot analysis. Proteomic analysis in present study suggests that DHBV infection induces perturbations in carbohydrate metabolism and cytoskeletal structure, which may favor host biosynthetic activities supporting viral replication and propagation. Glutamine metabolism is enhanced probably providing an alternative source for the generation of energy. Endoplasmic reticulum stress and oxidative stress response are found post the viral infection. The potential roles of some differentially expressed proteins such like GAPDH, beta-actin and annexin A2, were discussed. Taken together, the results of proteomic analysis from DHBV infection of PDHs will provide new clues for understanding the mechanisms during hepadnavirus infection. Further investigations into the roles of the differentially expressed cellular proteins will help to understand molecular mechanisms during hepadnavirus infection.Taken together, the present study explored global changes in PDHs of hepadnavirus infection by 2-DE analysis, using a natural PDHs-DHBV infection system. The proteomic analysis provides global insight to understanding of interactions between hepadnavirus and hepatocytes and molecular mechanisms of hepadnavirus pathogenesis.