Investigation of the protective effects of probiotics and polyphenols on polychlorinated biphenyl-induced inflammation and oxidative stress: impact of biotransformation

益生菌和多酚对多氯联苯诱导的炎症和氧化应激的保护作用的研究：生物转化的影响

• 批准号: RGPIN-2020-04901
• 负责人: Kubow, Stan
• 金额: $ 4.53万
• 依托单位: McGill University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2021
• 资助国家: 加拿大
• 起止时间: 2021-01-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=740282
• 关键词: Investigation; protective; effects; probiotics; polyphenols

Background: Polychlorinated biphenyls (PCBs) are ubiquitous and persistent toxic pollutants. Diet is the primary route of human PCB exposure, which is associated with gastrointestinal and nervous system damage. PCB toxicity could be mediated by gut microbiota dysbiosis and mitochondrial dysfunction. PCB toxicity mediated via oxidative stress might be protected by polyphenols that can induce an antioxidant response via nuclear factor erythroid 2-related factor 2 (Nrf2). Probiotics could be protective via alteration of gut microbiota and anti-inflammatory effects.      Research Progress: We developed gut digestion model that simulates digestion of the gastrointestinal (GI) tract including colonic reactors containing human microbiota. We showed that human microbial catabolism of common dietary polyphenols led to differing polyphenol metabolite profiles and antioxidant capacities. Caco-2/HepG2 cell co-cultures were used to mimic human intestinal and hepatic first pass metabolism of the colonic digests. PCB-126 and PCB-153, dominant congeners in diet and human plasma, were subjected to the GI model. The impact of digests of PCBs in the presence and absence of polyphenols and/or probiotics are being studied on: (a) profiles of human gut microbiota and their metabolites; and (b) mitochondrial function and oxidative injury in human intestinal epithelial cells. Objectives: The overall objectives are to combine GI model digests with the first pass metabolism co-culture system to: (1) identify microbial and first pass metabolites generated from PCB exposure in the study of mechanisms of PCB-induced toxicity on human intestinal and neural cells; and (2) assess the effects of microbial and first pass metabolites of polyphenols and probiotics on PCB metabolism and toxicity with a focus on oxidative stress, inflammation and the Nrf2 signalling pathway. Experimental Design and Methodology: GI model digests of parent PCBs produced in the presence and absence of polyphenols and/or probiotics are subjected to the first pass Caco-2/HepG2 co-culture system. PCB, polyphenol and probiotic microbial metabolites are assessed via GC-MS, LC-MS and HPLC.Microbiome analysis will be analyzed by real-time qPCR using Illumina MiSeq. The first pass metabolites will be applied onto normal human neural progenitor cells co-cultured with a model for the human blood brain barrier to assess neuronal differentiation and viability. Mitochondrial function will be studied by the MitoPlate assay. Cytokines will be assessed by Luminex. Nrf2 levels will be studied via Western blot analysis. Gene expression analysis will use RT2 Profiler PCR Array.  Significance: This project will provide insight of PCB toxicity as affected by gut digestive and first pass metabolic processes with special reference to how inflammation, oxidative injury and mitochondrial dysfunction can be mediated by probiotic and polyphenol intervention.

背景：多氯联苯（PCBs）是一种普遍存在的持久性有毒污染物。饮食是人体接触多氯联苯的主要途径，与胃肠道和神经系统损伤有关。多氯联苯毒性可能由肠道菌群失调和线粒体功能障碍介导。通过氧化应激介导的多氯联苯毒性可能受到多酚的保护，多酚可以通过核因子红细胞2相关因子2 （Nrf2）诱导抗氧化反应。益生菌可能通过改变肠道菌群和抗炎作用起到保护作用。研究进展：我们开发了模拟胃肠道（GI）消化的肠道消化模型，包括包含人类微生物群的结肠反应器。我们发现，常见膳食多酚的人体微生物分解代谢导致不同的多酚代谢物谱和抗氧化能力。Caco-2/HepG2细胞共培养用于模拟人肠道和肝脏结肠消化的首过代谢。多氯联苯126和多氯联苯153是饮食和人血浆中的优势同系物，采用GI模型。目前正在研究在多酚和/或益生菌存在和不存在的情况下多氯联苯的消化对以下方面的影响：(a)人类肠道微生物群及其代谢物的概况；(b)人肠上皮细胞线粒体功能和氧化损伤。目的：总体目标是将胃肠道模型消化与一次代代谢共培养系统相结合，以：(1)鉴定多氯联苯暴露产生的微生物和一次代代谢物，研究多氯联苯对人体肠道和神经细胞的毒性作用机制；(2)评估多酚和益生菌的微生物和第一次代谢物对多氯联苯代谢和毒性的影响，重点关注氧化应激、炎症和Nrf2信号通路。实验设计和方法：GI模型消化在存在和不存在多酚和/或益生菌的情况下产生的母体多氯联苯，进行第一次Caco-2/HepG2共培养系统。通过GC-MS、LC-MS和HPLC检测PCB、多酚和益生菌微生物代谢物。微生物组分析将使用Illumina MiSeq进行实时qPCR分析。第一代代谢物将应用于与人类血脑屏障模型共同培养的正常人类神经祖细胞上，以评估神经元的分化和活力。线粒体功能将通过MitoPlate试验进行研究。细胞因子将由Luminex评估。通过Western blot分析研究Nrf2水平。基因表达分析将使用RT2 Profiler PCR阵列。意义：本项目将深入了解多氯联苯毒性受肠道消化和首过代谢过程的影响，特别关注益生菌和多酚干预如何介导炎症、氧化损伤和线粒体功能障碍。