Department of Clinical and Biological Sciences, University of Turin, 10043, Orbassano (Turin), Italy.
Arch Biochem Biophys. 2020 Nov 15;694:108591. doi: 10.1016/j.abb.2020.108591. Epub 2020 Sep 19.
The alteration of the intestinal barrier function is currently believed to be involved in the pathogenesis of gut diseases mainly associated with the activation of inflammation processes. Diet plays an important role in the control of human gut integrity. Theobromine is a natural methylxanthine present in dark chocolate particularly abundant in cocoa bean shell. This is a polyphenol rich by-product generated in cocoa industrial processing, which is gaining value as a functional ingredient. This study aims to highlight for the first time the capability of theobromine in protecting the intestinal cell monolayer from a mixture of dietary oxysterols showing an inflammatory action in terms of IL-8 and MCP-1 overproduction. Differentiated CaCo-2 cells were treated with 60 μM oxysterol mixture and pre-incubated with 10 μM theobromine. Intestinal barrier damage was investigated in terms of tight junction claudin 1, occludin and JAM-A protein levels, matrix metalloproteinase (MMP) -2 and -9 activation and anti/pro-apoptotic protein changes. The observed cell monolayer permeability protection by theobromine may be due to its ability to inhibit the production of cytokines and MMPs that can be responsible for tight junction loss and apoptosis in intestinal cells. Our findings provide additional mechanistic hints on the healthy effect of theobromine cocoa component as an attractive natural molecule in the prevention of inflammatory gut diseases.
目前认为，肠道屏障功能的改变与主要与炎症过程激活相关的肠道疾病的发病机制有关。饮食在控制人体肠道完整性方面起着重要作用。可可碱是一种存在于黑巧克力中的天然甲基黄嘌呤，在可可豆壳中含量特别丰富。这是一种多酚丰富的副产物，在可可工业加工过程中产生，作为一种功能性成分越来越受到重视。本研究旨在首次强调可可碱保护肠道细胞单层免受具有炎症作用的膳食氧化固醇混合物的能力，这种混合物表现为 IL-8 和 MCP-1 过度产生。用 60μM 氧化固醇混合物处理分化的 CaCo-2 细胞，并预孵育 10μM 可可碱。根据紧密连接闭合蛋白 1、occludin 和 JAM-A 蛋白水平、基质金属蛋白酶 (MMP)-2 和 -9 的激活以及抗/促凋亡蛋白的变化来研究肠道屏障损伤。观察到的可可碱对细胞单层通透性的保护作用可能是由于其抑制细胞因子和 MMPs 产生的能力，这些细胞因子和 MMPs可能是导致肠道细胞紧密连接丧失和细胞凋亡的原因。我们的研究结果为可可碱作为一种有吸引力的天然分子在预防炎症性肠道疾病方面的健康效应提供了额外的机制线索。
