Inhibition of FoxO1 ameliorates hepatic steatosis and hepatitis in nonalcoholic steatohepatitis mice through regulation of gut microbiota

ObjectiveWe aimed to investigate the role of forkhead box O1 (FoxO1) inhibitor AS1842856 (AS) in nonalcoholic steatohepatitis (NASH) mice and the potential mechanisms.MethodsMice were given methionine-choline-sufficient (MCS), or methionine- and choline-deficient (MCD) diet for 5 weeks, along with AS (60 mg/kg) or vehicle gavage treatment (0.2 mL/day). Body and liver weight, serum triglyceride (TG), low-density lipoprotein-cholesterol (LDL-C), high-density lipoprotein-cholesterol (HDL-C), alanine aminotransferase (ALT), aspartate aminotransferase (AST), fasting glucose and insulin levels were measured. Liver macrophage infiltration and ileal ZO-1 protein expression were also detected. Interleukin (IL)-6, IL-1 beta, and tumor necrosis factor (TNF)-alpha, sterol regulatory element binding protein (SREBP)-1c, phosphoenolpyruvate carboxykinase (PEPCK), and glucose-6-phosphatase (G6Pase), alpha-smooth muscle actin (SMA), recombinant collagen type III alpha 1 (Col3a1), and connective tissue growth factor (Ctgf) expressions were measured. Stool samples were collected for 16S rDNA sequencing.ResultsCompared to the MCD group, AS attenuated liver weight, reduced serum TG, ALT, and AST levels, increased HDL-C levels, mitigated hepatic steatosis, decreased macrophage infiltration, and augmented ileal ZO-1 proteins in NASH mice. It also reduced the levels of IL-6, IL-1 beta, and TNF-alpha, alongside with the Srebp-1c mRNA expression. However, no significant effects on Pepck, G6Pase, alpha-SMA, Col3a1, or Ctgf were observed. Furthermore, AS promoted diversity and altered gut microbiota composition in NASH mice, causing increased beneficial bacteria like Akkermansia muciniphila, Parabacteroides distasonis, and Prevotellamassilia, which were associated with metabolic functions.ConclusionFoxO1 inhibitor AS ameliorated hepatic steatosis, inflammation, and intestinal dysbiosis in NASH mice, making it a potentially promising treatment for NASH. Forkhead protein O1 (FoxO1) inhibitor AS1842856 (AS) improves hepatic steatosis, reduces macrophage infiltration, lowers serum triglyceride (TG), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and increases high-density lipoprotein-cholesterol (HDL-C) levels in methionine- and choline-deficient (MCD) diet-induced nonalcoholic steatohepatitis (NASH) mice. It also reduces the levels of interleukin (IL)-6, IL-1 beta, and tumor necrosis factor (TNF)-alpha, alongside the sterol regulatory element binding protein (Srebp)-1c mRNA expression. Additionally, AS upregulates the expression of intestinal barrier protein ZO-1, enhances gut microbiota diversity, and boosts the abundance of beneficial bacteria such as Akkermansia muciniphila, Parabacteroides distasonis, and Prevotellamassilia. AS may be a promising candidate for the treatment of NASH.image