目的:我们旨在研究叉头盒O1(FoxO1)抑制剂AS1842856(AS)在非酒精性脂肪性肝炎(NASH)小鼠中的作用及其潜在机制。
方法:给予小鼠足够的蛋氨酸-胆碱(MCS),或甲硫氨酸和胆碱缺乏(MCD)饮食5周,以及AS(60mg/kg)或媒介物灌胃治疗(0.2mL/天)。身体和肝脏的重量,血清甘油三酯(TG),低密度脂蛋白胆固醇(LDL-C),高密度脂蛋白胆固醇(HDL-C),丙氨酸氨基转移酶(ALT),天冬氨酸转氨酶(AST),检测空腹血糖和胰岛素水平.同时检测肝巨噬细胞浸润和回肠ZO-1蛋白表达。白细胞介素(IL)-6,IL-1β,和肿瘤坏死因子(TNF)-α,甾醇调节元件结合蛋白(SREBP)-1c,磷酸烯醇丙酮酸羧激酶(PEPCK),和葡萄糖-6-磷酸酶(G6Pase),α-平滑肌肌动蛋白(SMA),重组III型胶原α1(Col3a1),测定结缔组织生长因子(Ctgf)表达。收集粪便样品用于16SrDNA测序。
结果:与MCD组相比,AS减轻肝脏重量,降低血清TG,ALT,AST水平,HDL-C水平升高,减轻肝脏脂肪变性,减少巨噬细胞浸润,和NASH小鼠回肠ZO-1蛋白增强。它还降低了IL-6,IL-1β的水平,和TNF-α,与Srebp-1cmRNA表达一起。然而,对Pepck没有显著影响,G6Pase,α-SMA,观察到Col3a1或Ctgf。此外,AS促进了NASH小鼠的多样性并改变了肠道菌群组成,导致有益细菌增加,如阿克曼西亚粘虫,双分支杆菌,和Prevotellamassilia,与代谢功能有关。
结论:FoxO1抑制剂AS改善肝脏脂肪变性,炎症,和NASH小鼠的肠道生态失调,使其成为NASH的潜在有希望的治疗方法。
OBJECTIVE: We aimed to investigate the role of forkhead box O1 (FoxO1) inhibitor AS1842856 (AS) in nonalcoholic steatohepatitis (NASH) mice and the potential mechanisms.
METHODS: Mice 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-1β, and tumor necrosis factor (TNF)-α, sterol regulatory element binding protein (SREBP)-1c, phosphoenolpyruvate carboxykinase (PEPCK), and glucose-6-phosphatase (G6Pase), α-smooth muscle actin (SMA), recombinant collagen type III α1 (Col3a1), and connective tissue growth factor (Ctgf) expressions were measured. Stool samples were collected for 16S rDNA sequencing.
RESULTS: Compared 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-1β, and TNF-α, alongside with the Srebp-1c mRNA expression. However, no significant effects on Pepck, G6Pase, α-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.
CONCLUSIONS: FoxO1 inhibitor AS ameliorated hepatic steatosis, inflammation, and intestinal dysbiosis in NASH mice, making it a potentially promising treatment for NASH.