Molecular Characterization of La Protein in a Mouse Acute Hepatitis Model / 中国药学杂志

ABSTRACT

OBJECTIVE: To explore the role of La protein in acute hepatitis B virus (HBV) by establishing a mouse model. METHODS: We constructed an acute hepatitis B virus model by hydrodynamics-based injection of plasmid pAAV/HBV1.2 containing 1.2 times HBV genome. The expression levels of HBV DNA, HBsAg and HBeAg in serum were detected by RT-PCR and ELISA, respectively. HBcAg in liver tissue were assayed by immunohistochemical staining.The level of ALT was detected by bioluminescence. Pathological changes in liver carried by Hematoxylin and eosin (HE) staining. RT-PCR, Western blot and ELISA were used to detect the changes of mRNA level and protein expression level of La protein. RESULTS: The levels of HBV DNA, HBsAg and HBeAg in the serum of mice, and the percentage of HBcAg-positive hepatocytes in liver tissues was up to 3.66% at day 5 showed that the mouse model of acute HBV was successfully constructed. The level of serum ALT and HE results showed that the liver damage was serious after the model was established, and then dropped sharplyand returned to normal. The mRNA level of La protein in the liver of the mice indicates that the level of La protein in the model group was generally higher than that in the control group. However, Western bolt results showed that there was no significant difference in intracellular protein levels, but the level of La protein in serum of mice is generally higher thancontrol group, especially on the 7th day (P<0.000 1). CONCLUSION: The acute HBV model was successfully constructed. The level of La protein in this model is related to HBV expression, suggesting that La protein may be involved in the pathogenesis of HBVandbea new marker in diagnosis of HBV.