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：多氯联苯是一种普遍存在的持久性有毒污染物。饮食是人类接触多氯联苯的主要途径，与胃肠道和神经系统损伤有关。多氯联苯的毒性可能通过肠道微生物区系失调和线粒体功能障碍来介导。多酚类物质可能通过核因子红系相关因子2(Nrf2)诱导抗氧化反应，从而保护氧化应激介导的多氯联苯毒性。益生菌可能通过改变肠道微生物区系和抗炎作用而起到保护作用。研究进展：我们开发了肠道消化模型，模拟胃肠道(GI)的消化，包括含有人类微生物区系的结肠反应器。我们发现，人类对常见饮食多酚的微生物分解代谢导致了不同的多酚代谢产物谱和抗氧化能力。Caco-2/HepG2细胞共培养模拟人肠道和肝脏结肠消化的首过代谢。饮食和人体血浆中的主要同系物多氯联苯-126和多氯联苯-153均符合GI模型。目前正在研究在存在和不存在多酚和/或益生菌的情况下多氯联苯的消化对以下方面的影响：(A)人类肠道微生物区系及其代谢物的分布；(B)人类肠道上皮细胞的线粒体功能和氧化损伤。目的：将GI模型消化与首过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首过代谢产物，以研究多氯联苯对人体肠道和神经细胞的毒性机制；(2)评估多酚和益生菌的微生物代谢产物和首过代谢产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：将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的GI模型消化置于第一代Caco-2/HepG2共培养系统中。多氯联苯、多酚和益生菌代谢产物通过GC-MS、LC-MS和HPLC进行评估。微生物组分析将使用Illumina MiSeq通过实时定量聚合酶链式反应进行分析。第一批代谢物将应用于与人血脑屏障模型共同培养的正常人类神经前体细胞，以评估神经元分化和活性。线粒体功能将通过有丝分裂板试验进行研究。细胞因子将由Luminex进行评估。NRF2水平将通过蛋白质印迹分析进行研究。基因表达分析将使用RT2 Profiler PCR阵列。意义：该项目将深入了解肠道消化和首次通过的代谢过程对多氯联苯的毒性，特别是如何通过益生菌和多酚干预来调节炎症、氧化损伤和线粒体功能障碍。