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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

基本信息

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：
2020
资助国家：
加拿大
起止时间：
2020-01-01 至 2021-12-31
项目状态：
已结题

来源：
https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=717453
关键词：
Investigation protective effects probiotics polyphenols

项目摘要

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）是普遍存在且持久的有毒污染物。饮食是人体接触 PCB 的主要途径，PCB 与胃肠道和神经系统损伤有关。 PCB 毒性可能是由肠道微生物群失调和线粒体功能障碍介导的。通过氧化应激介导的 PCB 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。益生菌可以通过改变肠道微生物群和抗炎作用来发挥保护作用。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群的结肠反应器。我们发现，人类微生物对常见膳食多酚的分解代谢导致了不同的多酚代谢谱和抗氧化能力。 Caco-2/HepG2 细胞共培养物用于模拟人类肠道和肝脏结肠消化物的首过代谢。 PCB-126 和 PCB-153 是饮食和人血浆中的主要同源物，接受了胃肠道模型的研究。正在研究多酚和/或益生菌存在和不存在时 PCB 消化物的影响： (a) 人类肠道微生物群及其代谢物的概况； (b) 人肠上皮细胞的线粒体功能和氧化损伤。目标：总体目标是将胃肠道模型消化物与首过代谢共培养系统相结合，以：（1）在研究 PCB 诱导的人类肠道和神经细胞毒性机制时，鉴定 PCB 暴露产生的微生物和首过代谢物； (2) 评估微生物以及多酚和益生菌的首过代谢物对 PCB 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：在存在和不存在多酚和/或益生菌的情况下产生的母体 PCB 的 GI 模型消化物经过第一遍 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。微生物组分析将使用 Illumina MiSeq 通过实时 qPCR 进行分析。首过代谢物将应用于与人血脑屏障模型共培养的正常人神经祖细胞，以评估神经元分化和活力。线粒体功能将通过 MitoPlate 测定进行研究。 Luminex 将评估细胞因子。 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 Array。意义：该项目将深入了解受肠道消化和首过代谢过程影响的 PCB 毒性，特别是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。