The Effects Of HBx Gene On Mouse Liver Inflammatory Factors And Cell Proliferation In Vivo

Objective: To explore the effect of HBx on liver inflammation factors and cell proliferation in KM mice and its potential mechanism,a stable expression model of HBx(Hepatitis B virus x)was established.Methods: The liver precursor cell line was infected by the lentivirus carrying HBx gene,and the infection was observed.The level of HBx in the HPCs was detected with q RT-PCR and Immunohistochemistry and Western blot.The HBx-EGFP-14-19 cells was injected into the portal vein of KM mice,livers were extracted 30 days,90 days,180 days and 360 days after treatment,and the level of HBx gene was detected.The expression of GFP was detected via using Fluorescence microscope.Western blot and q RT-PCR were used to detect the expression of Cyclin D1,CDK4,TNF?,IL-10 and the expression level of ERK1/2 signaling pathway members compared with the control groups.HBx-EGFP-14-19 mice feed for 180 days were injected with inhibitor U0126 for 11 days and then the livers of the mice was taken.The effects of U0126 on the expression of cyclin D1 and CDK4,inflammatory factors TNF?,and IL-10 were detected.Results: The construction of a mouse model stably expressing HBx gene is completed.Compared with the control group,Cyclin D1 and CDK4 transcription and protein levels were up-regulated.The level of TNF? was up-regulated.The level of IL-10 was reduced.The expression levels of Phosphorylated ERK1/2 and phosphorylated MEK1/2 were up-regulated.U0126,a MEK1/2 inhibitor,can significantly reverse the effects of HBx on the above-mentioned liver inflammation and cell cycle regulators.HE staining results showed that the mice in the experimental group had no obvious pathological changes in the liver at 30 days of culture;inflammatory cell mass infiltration around the veins and leaflets was clearly visible in the liver tissue at 90 days of culture;extensive liver cell fat was observed in the liver of the mice after 180 days of culture Degeneration,the cytoplasm contains circular vacuoles of varying sizes,and a small inflammatory cell infiltration spot can be seen locally.HBx-EGFP-14-19 360 days mouse liver tissue confluence area of lymphocyte infiltration;yellow arrow shows more hepatocyte edema,cytoplasm loose and light staining,some severe edema to balloon-like changes.Conclusions: HBx can cause liver inflammation and promote cell proliferation by activating ERK1 / 2 signal pathway abnormally in vivo.