Establishment of the Cellular Model of Chronic Hepatitis B Complicated with Non-alcoholic Fatty Liver Disease and the Intervention of Leontopodic Acid A

OBJECTIVE To establish the cellular model of chronic hepatitis B(CHB) complicated with non-alcoholic fatty liver disease(NAFLD), and to explore the correlations of CHB and NAFLD and the intervention of leontopodic acid A. METHODS Hepatitis B virus(HBV)‑transfected HepG2.2.15 hepatoma cells were induced by fat emulsion and oleic acid/palmitic acid(2︰1) mixture in stages to establish a model of CHB complicated with steatohepatitis. Triglyceride(TG) content and oil red O staining were employed to evaluate the degree of steatosis of HepG2.2.15 cells. The above model was employed to study the correlations of CHB and NAFLD by comparison of the levels of heme oxygenase-1(HO-1), tumor necrosis factor-α (TNF-α) and interleukin-1β(IL-1β) and HBV DNA in HepG2.2.15 cells before and after steatosis. The effect of leontopodic acid A on CHB complicated with NAFLD were evaluated by analyzing its effects on the viability, TG content and HBV DNA levels of HepG2.2.15 cells with steatosis. RESULTS The steatohepatitis in HepG2.2.15 cells was induced by 1% fat emulsion for 12 h and then by 0.5 mmol·L^-1 oleic acid/palmitic acid mixture for 24 h as the optimal modeling conditions for CHB complicated with NAFLD. Compared with before steatosis, the levels of HO-1, IL-1β and HBV DNA in HepG2.2.15 cells with steatosis were significantly decreased. Meanwhile, the expression of TNF-α was markedly increased. Additionally, at 10, 50 and 100 μg·mL^-1 of leontopodic acid A improved the viability against CHB complicated with NAFLD and decreased TG contents and HBV DNA levels significantly. CONCLUSION The induced fatty degeneration can interfere with the replication of HBV DNA. The expression changes of the related cytokines can be relevant to the inflammation that exists in the cells with fatty degeneration. This study preliminarily proves that leontopodic acid A has significant viability against CHB complicated with NAFLD.