Co-treatment of trigonelline and everolimus synergistically prevented chronic steatohepatitis induced by fast food diet and thioacetamide in a novel murine nonalcoholic steatohepatitis model.

OBJECTIVES: Nonalcoholic steatohepatitis (NASH) is a strong risk factor for end-stage liver disease. Trigonelline (TG) is a plant alkaloid with anti-oxidant, anti-dyslipidemic, and anti-insulin resistance activities. Everolimus (EV), a conventional drug and an mTOR inhibitor, has been demonstrated to improve metabolic outcomes. The synergistic effect of the co-treatment of TG and EV against NASH conditions remains unknown. MATERIALS AND METHODS: We have developed a fast food (FF)-diet and thioacetamide-induced chronic steatohepatitis in a C57BL/6J mice model of 24 weeks duration. We have evaluated the synergistic protective effect of TG and EV at reduced doses to avoid any undesired toxic manifestations of the FF and thioacetamide. The study was demonstrated by comparative analysis across different groups after 24 weeks. RESULTS: Co-exposure to FF diet and thioacetamide resulted in chronic steatohepatitis, evident by focal necrosis, bridging fibrosis, loss of liver architecture, and excessive collagen deposition. Protein and gene analysis revealed enhanced de novo lipogenesis (SREBP-1, PPAR-Ƴ, CD36), inflammation (interleukin-6, tumor necrosis factor, CYP2E1), fibrosis (transforming growth factor beta, alpha-smooth muscle actin, tissue inhibitors of metalloproteinases-1), and extracellular matrix deposition (MMP-1, Col1A1). TG + EV at reduced doses showed marked synergistic effects in preventing inflammation, fibrosis, and lipogenesis markers. CONCLUSION: This study provides a novel 24-week FF diet and thioacetamide-induced murine NASH model for possible preclinical drug discovery studies. Furthermore, our treatment regimen discovered the synergistic effect of TG and EV at reduced doses in preventing chronic steatohepatitis.