Gene regulation of intestinal porcine epithelial cells IPEC-J2 is dependent on the site of deoxynivalenol toxicological action.
Gene regulation of intestinal porcine epithelial cells IPEC-J2 is dependent on the site of deoxynivalenol toxicological action.
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肠道猪上皮细胞的基因调节IPEC-J2取决于脱氧基因维烯醇毒理学作用部位。
DOI：
10.1371/journal.pone.0034136
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Kahlert S
中科院分区：
文献类型：
--
作者：
Diesing AK;Nossol C;Ponsuksili S;Wimmers K;Kluess J;Walk N;Post A;Rothkötter HJ;Kahlert S
肠上皮细胞层代表肠道腔侧和全身侧之间的边界。物质的吸收和排除之间的决定是肠道的基本功能，并且沿着肠轴变化。因此，潜在的有毒物质可能通过血流到达上皮细胞层的基底外侧区域。霉菌毒素脱氧雪腐镰刀菌烯醇 (DON) 是镰孢菌衍生的次级代谢产物，已知会进入血流，并在体外对极化上皮细胞层的基底外侧表现出惊人的毒性。在这里，我们分析了基因表达中反映的顶端和基底外侧 DON 毒性的潜在机制。我们使用空肠来源的极化肠猪上皮细胞系 IPEC-J2 作为体外细胞培养模型。通过从膜插入物的顶端或基底外侧室施加 DON 72 小时来模拟上皮细胞层的管腔和全身 DON 攻击。我们将未经处理和 DON 处理的 IPEC-J2 细胞的全基因组基因表达与 Affymetrix 的 GeneChip® 猪基因组阵列进行了比较。尽管通过跨上皮电阻测量，受攻击的细胞层完好无损，但与相应的顶端应用相比（2539 与 267），低基底外侧 DON（200 ng/mL）应用触发了 10 倍多的基因转录本。通过生物信息资源 DAVID 对受调节基因的分析，确定了受 DON 应用浓度和方向调节的几组生化途径。通过定量 PCR 详细分析代表细胞代谢、信息处理和结构设计途径的所选基因。我们的研究结果清楚地表明，上皮细胞层的顶端和基底外侧挑战触发不同的基因反应谱，同时对基底外侧挑战具有更高的敏感性。霉菌毒素毒理学潜力的评估应考虑到依赖于应用途径的基因调控差异。
The intestinal epithelial cell layer represents the border between the luminal and systemic side of the gut. The decision between absorption and exclusion of substances is the quintessential function of the gut and varies along the gut axis. Consequently, potentially toxic substances may reach the basolateral domain of the epithelial cell layer via blood stream. The mycotoxin deoxynivalenol (DON) is a Fusarium derived secondary metabolite known to enter the blood stream and displaying a striking toxicity on the basolateral side of polarised epithelial cell layers in vitro. Here we analysed potential mechanisms of apical and basolateral DON toxicity reflected in the gene expression. We used the jejunum-derived, polarised intestinal porcine epithelial cell line IPEC-J2 as an in vitro cell culture model. Luminal and systemic DON challenge of the epithelial cell layer was mimicked by a DON application from the apical or basolateral compartment of membrane inserts for 72 h. We compared the genome-wide gene expression of untreated and DON-treated IPEC-J2 cells with the GeneChip® Porcine Genome Array of Affymetrix. Low basolateral DON (200 ng/mL) application triggered 10 times more gene transcripts in comparison to the corresponding apical application (2539 versus 267) despite the intactness of the challenged cell layer as measured by transepithelial electrical resistance. Analysis of the regulated genes by bioinformatic resource DAVID identified several groups of biochemical pathways modulated by concentration and orientation of DON application. Selected genes representing pathways of the cellular metabolism, information processing and structural design were analysed in detail by quantitative PCR. Our findings clearly show that apical and basolateral challenge of epithelial cell layers trigger different gene response profiles paralleled with a higher susceptibility towards basolateral challenge. The evaluation of toxicological potentials of mycotoxins should take this difference in gene regulation dependent on route of application into account.
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影响因子：
4.4
作者：
Iwahashi H;Kitagawa E;Suzuki Y;Ueda Y;Ishizawa YH;Nobumasa H;Kuboki Y;Hosoda H;Iwahashi Y
通讯作者：
Iwahashi Y
影响因子：
2.3
作者：
Schierack, P;Nordhoff, M;Wieler, LH
通讯作者：
Wieler, LH
影响因子：
3.5
作者：
Diesing, Anne-Kathrin;Nossol, Constanze;Kahlert, Stefan
通讯作者：
Kahlert, Stefan
影响因子：
--
作者：
RHOADS, JM;CHEN, W;PARADISO, AM
通讯作者：
PARADISO, AM
影响因子：
3.8
作者：
Shi, Yuhui;Porter, Katie;Pestka, James J.
通讯作者：
Pestka, James J.