Laparra J M, Sanz Y
Instituto de Agroquímica y Tecnología de Alimentos (CSIC), Burjassot (Valencia), Spain.
J Cell Biochem. 2010 Mar 1;109(4):801-7. doi: 10.1002/jcb.22459.
Celiac disease (CD) is a chronic enteropathy triggered by intake of gliadin, the toxic component of gluten. This study aims at evaluating the capacity of different Bifidobacterium strains to counteract the inflammatory effects of gliadin-derived peptides in intestinal epithelial (Caco-2) cells. A commercial extract of several gliadin (Gld) types (alpha, beta, gamma, [symbol: see text] ) was subjected to in vitro gastrointestinal digestion (pepsin at pH 3, pancreatin-bile at pH 6), inoculated or not with cell suspensions (10(8) colony forming units/ml) of either B. animalis IATA-A2, B. longum IATA-ES1, or B. bifidum IATA-ES2, in a bicameral system. The generated gliadin-derived peptides were identified by reverse phase-HPLC-ESI-MS/MS. Caco-2 cell cultures were exposed to the different gliadin peptide digestions (0.25 mg protein/ml), and the mRNA expression of nuclear factor kappa-B (NF-kappaB), tumor necrosis factor alpha (TNF-alpha), and chemokine CXCR3 receptor were analyzed by semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) in stimulated cells. The production of the pro-inflammatory markers NF-kappaB p50, TNF-alpha, and IL-1beta (interleukine 1beta) by Caco-2 cells was also determined by ELISA. The peptides from gliadin digestions inoculated with bifidobacteria did not exhibit the toxic amino acid sequences identified in those noninoculated (alpha/beta-Gld [158-164] and alpha/beta-Gld [122-141]). The RT-PCR analysis evidenced a down-regulation in mRNA expression of pro-inflammatory biomarkers. Consistent with these results the production of NF-kappaB, TNF-alpha, and IL-1beta was reduced (18.2-22.4%, 28.0-64.8%, and abolished, respectively) in cell cultures exposed to gliadin digestions inoculated with bifidobacteria. Therefore, bifidobacteria change the gliadin-derived peptide pattern and, thereby, attenuate their pro-inflammatory effects on Caco-2 cells.
乳糜泻（CD）是一种由摄入麦醇溶蛋白（麸质的有毒成分）引发的慢性肠病。本研究旨在评估不同双歧杆菌菌株对抗麦醇溶蛋白衍生肽对肠上皮（Caco-2）细胞炎症作用的能力。将几种类型（α、β、γ、[符号：见原文]）的麦醇溶蛋白（Gld）的商业提取物进行体外胃肠道消化（pH 3时用胃蛋白酶，pH 6时用胰酶 - 胆汁），在双室系统中接种或不接种动物双歧杆菌IATA - A2、长双歧杆菌IATA - ES1或两歧双歧杆菌IATA - ES2的细胞悬液（10⁸ 菌落形成单位/毫升）。通过反相 - HPLC - ESI - MS/MS鉴定产生的麦醇溶蛋白衍生肽。将Caco - 2细胞培养物暴露于不同的麦醇溶蛋白肽消化物（0.25毫克蛋白质/毫升），并通过半定量逆转录聚合酶链反应（RT - PCR）分析刺激细胞中核因子κB（NF - κB）、肿瘤坏死因子α（TNF - α）和趋化因子CXCR3受体的mRNA表达。还通过ELISA测定Caco - 2细胞产生促炎标志物NF - κB p50、TNF - α和白细胞介素1β（IL - 1β）的情况。接种双歧杆菌的麦醇溶蛋白消化物中的肽未表现出在未接种的消化物中鉴定出的有毒氨基酸序列（α/β - Gld [158 - 164]和α/β - Gld [122 - 141]）。RT - PCR分析证明促炎生物标志物的mRNA表达下调。与这些结果一致，在暴露于接种双歧杆菌的麦醇溶蛋白消化物的细胞培养物中，NF - κB、TNF - α和IL - 1β的产生减少（分别为18.2 - 22.4%、28.0 - 64.8%和完全消除）。因此，双歧杆菌改变了麦醇溶蛋白衍生肽的模式，从而减弱了它们对Caco - 2细胞的促炎作用。
