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炎症刺激下肠上皮细胞对富含类胡萝卜素和多酚的体外消化李子和卷心菜的蛋白质组学反应。

Proteomic response of inflammatory stimulated intestinal epithelial cells to in vitro digested plums and cabbages rich in carotenoids and polyphenols.

作者信息

Kaulmann Anouk, Planchon Sébastien, Renaut Jenny, Schneider Yves-Jacques, Hoffmann Lucien, Bohn Torsten

机构信息

Luxembourg Institute of Science and Technology - Environmental Research and Innovation Department, L-4422 Belvaux, Luxembourg.

Institut des Sciences de la Vie, UCLouvain, B-1348 Louvain-la-Neuve, Belgium.

出版信息

Food Funct. 2016 Oct 12;7(10):4388-4399. doi: 10.1039/c6fo00674d.

Abstract

Due to their anti-oxidant and anti-inflammatory potential, polyphenol and carotenoid-rich plant foods have been suggested as promising phytochemicals in the prevention of or as adjuvants regarding inflammatory bowel diseases (IBD). In the present study, we investigated whether plum (Italian Plum, Prunus cocomilla), or cabbage (Kale, Brassica oleracea var. sabellica), selected for their high phytochemical content, are able to reduce inflammation in cellular models of the intestinal epithelium, employing proteomic methods. For this purpose, plum/cabbage (carotenoid content: 1.9 mg per 100 g resp. 13 mg per 100 g; polyphenol content: 83 mg per 100 g resp. 27 mg per 100 g) were gastro-intestinally digested, and aliquots exposed (18 h) to either a monoculture (Caco-2) or a triple culture (Caco-2/HT-29-MTX (90 : 10, v/v) with THP-1 like macrophages), stimulated (with LPS, TNF-α, and IL-1β) to induce inflammation. Cells (Caco-2, Caco-2/HT-29-MTX, and THP-1) were then harvested separately, and proteomic analyses of total cell extracts were carried out by 2D-DIGE. In the monoculture, 68 protein-spots were significantly (p < 0.05, expression ratio >1.5) differentially regulated due to the Kale and Italian plum digesta, and in the co-culture 206 protein-spots, compared to digesta without plum/cabbage. These belonged to 27 (monoculture) and 76 (coculture) uniquely identified proteins, suggesting the coculture to be a more sensitive model. Proteins included antioxidant enzymes such as catalase, superoxide dismutase and glutathione-S-transferases. Only 3 proteins were differentially regulated in the THP-1 cells, perhaps as these were only indirectly exposed. The results show promise regarding some aspects related to IBD complications, however, employing phytochemical-rich food items should be further investigated in in vivo trials.

摘要

由于其抗氧化和抗炎潜力,富含多酚和类胡萝卜素的植物性食物被认为是预防炎症性肠病(IBD)或作为其辅助治疗的有前景的植物化学物质。在本研究中,我们利用蛋白质组学方法研究了因富含植物化学物质而被挑选出的李子(意大利李子,欧洲李)或卷心菜(羽衣甘蓝,甘蓝变种)是否能够减轻肠上皮细胞模型中的炎症。为此,将李子/卷心菜(类胡萝卜素含量:每100克分别为1.9毫克和13毫克;多酚含量:每100克分别为83毫克和27毫克)进行胃肠道消化,取等分试样暴露(18小时)于单培养物(Caco-2)或三重培养物(Caco-2/HT-29-MTX(90 : 10,v/v)与类似THP-1的巨噬细胞)中,用脂多糖(LPS)、肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β)刺激以诱导炎症。然后分别收获细胞(Caco-2、Caco-2/HT-29-MTX和THP-1),并通过二维差异凝胶电泳(2D-DIGE)对总细胞提取物进行蛋白质组分析。在单培养物中,与不含李子/卷心菜的消化物相比,羽衣甘蓝和意大利李子消化物导致68个蛋白点有显著差异调节(p < 0.05,表达比>1.5),在共培养物中有206个蛋白点有差异调节。这些属于27个(单培养物)和76个(共培养物)唯一鉴定的蛋白质,表明共培养物是更敏感的模型。蛋白质包括抗氧化酶,如过氧化氢酶、超氧化物歧化酶和谷胱甘肽-S-转移酶。在THP-1细胞中只有3种蛋白质有差异调节,可能是因为这些细胞只是间接暴露。结果在与IBD并发症相关的某些方面显示出前景,然而,富含植物化学物质的食物在体内试验中应进一步研究。

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