São Paulo State University (UNESP), Botucatu Medical School, Department of Pathology, Botucatu, Brazil; São Paulo State University (UNESP), Biosciences Institute, Department of Structural and Functional Biology, Botucatu, Brazil.
São Paulo State University (UNESP), Biosciences Institute, Department of Structural and Functional Biology, Botucatu, Brazil; Federal University of Grande Dourados (UFGD), Dourados, MS, Brazil.
Nutrition. 2022 Nov-Dec;103-104:111836. doi: 10.1016/j.nut.2022.111836. Epub 2022 Aug 29.
Non-alcoholic fatty liver disease (NAFLD) has a growing epidemiologic and economic burden. It is associated with Western diet (WD) patterns, and its pathogenesis involves metabolic disorders (obesity, dyslipidemia, hyperglycemia, and diabetes) and gut dysbiosis, features that are usually neglected or not reproduced by most animal models. Thus, we established a 6-mo WD-induced NAFLD mouse model associated with metabolic disorder, investigating its main features at the gut microbiome-liver-adipose tissue axis, also evaluating the correlations of gut dysbiosis to the other disease outcomes.
Male C57 BL6 mice received a high-fat (30% lard and 0.2% cholesterol, ∼57% calories) and sucrose-rich (20%) chow, and a high-sugar solution (23.1 and 18.9 g/L of D-fructose and D-glucose) for 6 mo.
The model featured high serum cholesterol levels, glucose intolerance, and hyperinsulinemia. WD intervention resulted in extensive macro/microvesicular liver steatosis and pericellular fibrosis-resembling human disease-accompanied by hepatic stellate cell activation and CD68+ macrophage infiltration, increased protein levels of proinflammatory p65-nuclear factor-κB, interleukin-6 and tumor necrosis factor-α, with decreased antioxidant regulator Nrf2. Mice showed clear obesity with adipocyte hypertrophy, and CD68+macrophage/mast cell infiltration in adipose tissue while a reduction in number of goblet cells was also observed in the small intestine. Moreover, the pyrosequencing of the 16 S ribosomal RNA of gut cecal content showed decreased bacterial diversity, enriched Firmicutes and Proteobacteria, decreased Bacteroidetes and Fusobacteria, and increased ratio of Firmicutes to Bacteroidetes. Bacteroidetes and Bacteroides had the highest number of significant correlations with liver-adipose tissue axis outcomes. In silico analysis of gut microbiome in NAFLD obese patients revealed a depletion in Bacteroides, which also correlated to disease outcomes.
This mice model gathered suitable phenotypical alterations in gut-liver-adipose tissue axis that resembled NAFLD associated with metabolic disorders in humans and may be considered for preclinical investigation.
非酒精性脂肪性肝病（NAFLD）的流行病学和经济负担日益加重。它与西方饮食（WD）模式有关，其发病机制涉及代谢紊乱（肥胖、血脂异常、高血糖和糖尿病）和肠道菌群失调，这些特征通常被大多数动物模型所忽视或无法重现。因此，我们建立了一个 6 个月的 WD 诱导的 NAFLD 小鼠模型，该模型与代谢紊乱有关，研究了其在肠道微生物组-肝脏-脂肪组织轴上的主要特征，同时还评估了肠道菌群失调与其他疾病结果的相关性。
雄性 C57BL6 小鼠接受高脂肪（30%猪油和 0.2%胆固醇，约 57%热量）和富含蔗糖（20%）的饲料，以及高糖溶液（23.1 和 18.9 g/L 的 D-果糖和 D-葡萄糖），持续 6 个月。
该模型表现出高血清胆固醇水平、葡萄糖不耐受和高胰岛素血症。WD 干预导致广泛的大/微泡性肝脂肪变性和细胞周纤维化，类似于人类疾病，伴有肝星状细胞激活和 CD68+巨噬细胞浸润，促炎 p65-核因子-κB、白细胞介素-6 和肿瘤坏死因子-α的蛋白水平升高，抗氧化调节剂 Nrf2 减少。小鼠表现出明显的肥胖，脂肪细胞肥大，脂肪组织中 CD68+巨噬细胞/肥大细胞浸润，同时小肠中杯状细胞数量减少。此外，肠道盲肠内容物 16S 核糖体 RNA 的焦磷酸测序显示细菌多样性减少，厚壁菌门和变形菌门富集，拟杆菌门和梭杆菌门减少，厚壁菌门与拟杆菌门的比例增加。拟杆菌门和拟杆菌属与肝-脂肪组织轴结果的相关性最高。对 NAFLD 肥胖患者肠道微生物组的计算分析显示，拟杆菌属减少，这也与疾病结果相关。
该小鼠模型聚集了在肠道-肝脏-脂肪组织轴上适合表型改变的 NAFLD，与人类代谢紊乱相关的 NAFLD 相似，可用于临床前研究。
