Food Nutrition and Health Program, Faculty of Land and Food Systems, The University of British Columbia, 2205 East Mall, Vancouver, V6T-1Z4, B.C, Canada.
Food Nutrition and Health Program, Faculty of Land and Food Systems, The University of British Columbia, 2205 East Mall, Vancouver, V6T-1Z4, B.C, Canada.
Arch Biochem Biophys. 2024 Jun;756:109978. doi: 10.1016/j.abb.2024.109978. Epub 2024 Apr 16.
A 2D-intestinal epithelial Caco-2/RAW 264.7 macrophage co-culture model was developed to demonstrate the relative efficacy of different phenolic acids to mitigate changes in Caco-2 epithelial cell redox state initiated both directly by autoxidation products, HO, and indirectly through cell communication events originating from cytokine stimulated macrophage. An inducer cocktail (lipopolysaccharide + interferon gamma) was used to activate RAW 264.7 cells in the 2D- Caco-2/RAW co-culture and intracellular changes in Caco-2 cell redox signaling occurred in response to positive changes (p < 0.05) in inflammatory biomarkers derived in macrophage that included IL-6, TNF-α, nitric oxide and peroxynitrite, respectively. Phenolic acids varied in relative capacity to reduce NF-κB (nuclear factor kappa-light-chain-enhancer of activated B cells) in cocktail inflamed induced macrophage. This response in addition to the relative predisposition of gallic acid (GA) to undergo autoxidation to generate HO activity (p < 0.05), culminated in downstream cell signaling in Caco-2 nuclear factor erythroid 2-related factor (Nrf2) activity (increase 26.9 %), altered monolayer integrity (increase 33.7 %), and release of interleukin 8 (IL-8) (decrease 80.5 %) (p < 0.05). It can be concluded that the co-culture model described herein was useful to assess the importance of communication between cytokine stimulated macrophage and intestinal cells. Moreover, the relative unique efficacy of GA, compared to other phenolic acids tested to protect against activated macrophage induced changes related to intestinal dysfunction were particularly relevant to epithelial redox signaling, intestinal permeability and regulation of tight junction proteins. This study concludes that phenolic acids are not equal in the capacity to protect against intestinal cell dysfunction despite some indication of biological activity.
建立了一种 2D 肠上皮 Caco-2/RAW264.7 巨噬细胞共培养模型，以证明不同酚酸缓解由自氧化产物 HO 直接引发以及通过源自细胞因子刺激巨噬细胞的细胞通讯事件间接引发的 Caco-2 上皮细胞氧化还原状态变化的相对功效。诱导鸡尾酒（脂多糖+干扰素 γ）用于激活 2D-Caco-2/RAW 共培养中的 RAW264.7 细胞，Caco-2 细胞氧化还原信号转导的细胞内变化发生在响应于源自巨噬细胞的炎症生物标志物的阳性变化（p<0.05）时，这些标志物包括分别为白细胞介素 6（IL-6）、肿瘤坏死因子-α（TNF-α）、一氧化氮和过氧亚硝酸盐。酚酸在还原 NF-κB（核因子 kappa-轻链增强子的激活 B 细胞）的相对能力方面存在差异，在诱导的巨噬细胞炎症中。除了没食子酸（GA）相对容易自氧化产生 HO 活性的倾向（p<0.05）外，这种反应最终导致 Caco-2 核因子红细胞 2 相关因子（Nrf2）活性的下游细胞信号转导（增加 26.9%）、单层完整性改变（增加 33.7%）和白细胞介素 8（IL-8）释放（减少 80.5%）（p<0.05）。可以得出结论，本文所述的共培养模型可用于评估细胞因子刺激的巨噬细胞与肠细胞之间通讯的重要性。此外，GA 与其他测试的酚酸相比具有相对独特的功效，可保护免受激活的巨噬细胞诱导的与肠道功能障碍相关的变化，这与上皮细胞氧化还原信号转导、肠道通透性和紧密连接蛋白的调节特别相关。本研究得出的结论是，尽管酚酸具有一定的生物活性，但它们在保护肠道细胞功能障碍方面的能力并不相同。
