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（PCB）是普遍存在的持续有毒污染物。饮食是人类PCB暴露的主要途径，与胃肠道和神经系统损害有关。 PCB毒性可以通过肠道菌群营养不良和线粒体功能障碍来介导。通过氧化应激介导的PCB毒性可能受到可以通过核因子2相关因子2（NRF2）诱导抗氧化反应的多酚保护。益生菌可以通过改变肠道菌群和抗炎作用来保护。研究进度：我们开发了肠道消化模型，以模拟胃肠道（GI）的消化，包括含有人类微生物群的结肠反应器。我们表明，普通饮食多酚的人类微生物分解代谢导致了不同的多酚代谢物谱和抗氧化能力。 CACO-2/HEPG2细胞共培养被用来模仿结肠摘要的人类肠道和肝炎首次通过代谢。 PCB-126和PCB-153（饮食和人血浆中的主要同源物）受到了GI模型的约束。在存在和不存在多酚和/或益生菌的情况下，PCB的摘要的影响正在研究：（a）人类肠道微生物群及其代谢产物的剖面； （b）人肠上皮细胞中的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第一个通过代谢共培养系统相结合：（1）在研究PCB诱导的人类肠道和神经细胞的毒性的研究中，识别由PCB暴露产生的微生物和第一次通过代谢物； （2）评估多酚和益生菌对PCB代谢和毒性的微生物和第一次通过代谢物的影响，重点是氧化应激，炎症和NRF2信号传导途径。实验设计和方法论：在存在和不存在多酚和/或益生菌的情况下产生的父型PCB的GI模型消化受到第一次通过CACO-2/HEPG2共培养系统。 PCB，多酚和益生菌微生物代谢物通过GC-MS，LC-MS和HPLC进行评估。将使用Illumina Miseq通过实时QPCR分析Microbiome分析。第一个通过代谢物将被应用于与人类血脑屏障的模型共培养的正常人神经元基因细胞，以评估神经元分化和生存能力。线粒体功能将由线条板测定法进行研究。细胞因子将通过Luminex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：该项目将提供PCB毒性的洞察力，并受到肠道消化的影响和首次通过代谢过程，并特别提及如何通过益生菌和多酚干预来介导感染，氧化损伤和线粒体功能障碍。