Alterations in secondary gut metabolites derived from the microbial fermentation of food in the gut have significant effects on various aspects of host physiology. Our recent studies on obese mice treated with Orlistat, an antiobesity treatment, revealed a significantly altered gut microbial profile marked by an over-abundance of Proteobacteria and alterations in secondary gut metabolites. In this study, we determined effect of fecal metabolites from high-fat diet fed mice treated with Orlistat (HFDOrl) on colonic epithelial cells in relation to inflammation, barrier function, mitochondrial activity, reactive oxygen species (ROS) levels, and oxidative stress. Quantitative PCR was used to measure intestinal mRNA expression of oxidative stress, inflammation, apoptosis, and gut barrier function genes in mice on a high fat diet with and without Orlistat treatment versus those fed a low-fat diet (HFDOrl, HFD, Normal diet-fed [ND] respectively). Alterations to antioxidant function in HCT-116-ARE-luciferase stable cell line and mitochondrial function in Caco-2 cells was analyzed under oxidative stress with exposure to aqueous fecal extracts from HFDOrl, HFD, and ND groups. The results of this study indicate that a significant increase in anti-oxidative response was observed based on the luciferase activity of HCT-116-ARE-luciferase stable cells. Increased maximal respiration and mitochondrial ROS under oxidative stress was also detected in confluent Caco-2 cells resulting from exposure to fecal extracts from the HFDOrl group compared with the HFD group and pure Orlistat. Furthermore, mice from the HFDOrl group exhibited a significant increase in colonic epithelial expression of oxidative markers (Nrf-2 and SOD-2), inflammation-related markers (IL-6 and TNF-α), and gut barrier function markers (Muc-2 and Occludin). Taken together, the results suggest that Orlistat treatment in the HFD group causes changes in secondary gut metabolites which affect the colonic redox state and may eventually lead to the development of inflammatory, oxidative, and mitochondrial dysfunction at the cellular level.

中文翻译：

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奥利司他是一种竞争性脂肪酶抑制剂，用作抗肥胖药物，可增强肠道的炎症反应

源自肠道中食物微生物发酵的次生肠道代谢物的变化对宿主生理的各个方面都有显着影响。我们最近对使用奥利司他（一种抗肥胖治疗）治疗的肥胖小鼠进行的研究显示，肠道微生物特征显着改变，其特征是变形杆菌过多和肠道次生代谢物的改变。在这项研究中，我们确定了用奥利司他 (HFDOrl) 治疗的高脂饮食喂养小鼠的粪便代谢物对结肠上皮细胞的炎症、屏障功能、线粒体活性、活性氧 (ROS) 水平和氧化应激的影响。定量PCR用于测量氧化应激、炎症、细胞凋亡、与喂食低脂肪饮食（分别为HFDOrl、HFD、正常饮食喂养[ND]）的小鼠相比，高脂饮食和不使用奥利司他治疗的小鼠的肠道屏障功能基因和肠道屏障功能基因。在暴露于来自 HFDOr1、HFD 和 ND 组的水性粪便提取物的氧化应激下，分析了 HCT-116-ARE-荧光素酶稳定细胞系中抗氧化功能和 Caco-2 细胞中线粒体功能的改变。该研究的结果表明，基于 HCT-116-ARE-荧光素酶稳定细胞的荧光素酶活性，观察到抗氧化反应显着增加。与 HFD 组和纯奥利司他相比，由于暴露于 HFDOrl 组的粪便提取物，在汇合的 Caco-2 细胞中也检测到在氧化应激下最大呼吸和线粒体 ROS 的增加。此外，HFDOrl 组小鼠结肠上皮表达的氧化标志物（Nrf-2 和 SOD-2）、炎症相关标志物（IL-6 和 TNF-α）和肠道屏障功能标志物（Muc-2 和闭塞蛋白）。总之，结果表明，在 HFD 组中使用奥利司他治疗会导致肠道次级代谢物发生变化，这些代谢物会影响结肠氧化还原状态，并可能最终导致细胞水平上炎症、氧化和线粒体功能障碍的发展。