Transcriptional response of cultured porcine intestinal epithelial cells to micro algae extracts in the presence and absence of enterotoxigenic Escherichia coli.

Genes & Nutrition Pub Date : 2019-03-19 eCollection Date: 2019-01-01 DOI:10.1186/s12263-019-0632-z
Marcel Hulst, Rommie van der Weide, Arjan Hoekman, Marinus van Krimpen
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引用次数: 7

Abstract

Background: Micro algae's are worldwide considered as an alternative source of proteins in diets for animals and humans. Micro algae also produce an array of biological active substances with potential to induce beneficial and health promoting effects. To better understand the mode of action of micro algae's when applied as additive in diets, porcine intestinal epithelial cells (IPEC-J2), stressed by enterotoxigenic Escherichia coli (ETEC) or under non-stressed conditions, were exposed to micro algae extracts and changes in gene expression were recorded.

Methods: IPEC-J2 cells were exposed for 2 and 6 h to extracts prepared from the biomass of the microalgae Chlorella vulgaris (C), Haematococcus pluvialis (H), Spirulina platensis (S), or a mixture of Scenedesmus obliques and Chlorella sorokiniana (AM), in the absence and presence of ETEC. Gene expression in cells was measured using porcine "whole genome" microarrays.

Results: The micro algae extracts alone enhanced the expression of a set of genes coding for proteins with biological activity that are secreted from cells. These secreted proteins (hereafter denoted as effector proteins; EPs) may regulate processes like remodelling of the extracellular matrix, activation of an antiviral/bacterial response and oxygen homeostasis in the intestine and periphery. Elevated gene expression of immunostimulatory proteins CCL17, CXCL2, CXCL8 (alias IL8), IFNA, IFNL1, HMOX1, ITGB3, and THBS1 was observed in response to all four extracts in the absence or presence of ETEC. For several of these immunostimulatory proteins no elevated expression was observed when cells were exposed to ETEC alone. Furthermore, all extracts highly stimulated expression of an antisense RNA of the mitochondrial/peroxisome symporter SLC25A21 gene in ETEC-challenged cells. Inhibition of SLC25A21 translation by this antisense RNA may impose a concentration gradient of 2-oxoadipic and 2-oxoglutarate, both metabolites of fatty acid β-oxidation, between the cytoplasm and the interior of these organelles.

Conclusions: Exposure of by ETEC stressed intestinal epithelium cells to micro algae extracts affected "fatty acid β-oxidation", ATP and reactive oxygen species production and (de) hydroxylation of lysine residues in procollagen chains in these cells. Elevated gene expression of specific EPs and immunostimulatory proteins indicated that micro algae extracts, when used as feed/food additive, can steer an array of metabolic and immunological processes in the intestines of humans and monogastric animals stressed by an enteric bacterial pathogen.

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在存在和不存在产肠毒素大肠杆菌的情况下,培养的猪肠上皮细胞对微藻提取物的转录反应。
背景:微藻在世界范围内被认为是动物和人类饮食中蛋白质的替代来源。微藻还能产生一系列生物活性物质,具有潜在的有益和促进健康的作用。为了更好地了解微藻作为饲料添加剂的作用模式,本研究将猪肠上皮细胞(IPEC-J2)置于产肠毒素大肠杆菌(ETEC)应激和非应激条件下,记录微藻提取物对其基因表达的影响。方法:将IPEC-J2细胞分别暴露于小球藻(Chlorella vulgaris, C)、雨生红球菌(Haematococcus pluvialis, h)、螺旋藻(Spirulina platensis, S)的生物提取物中,或斜状小球藻(Scenedesmus oblique)和小球藻(Chlorella sorokiniana, AM)的混合物中,分别在ETEC不存在和ETEC存在的情况下,培养2和6 h。使用猪“全基因组”微阵列测量细胞中的基因表达。结果:微藻提取物增强了细胞分泌的一组具有生物活性的蛋白质编码基因的表达。这些分泌的蛋白质(以下称为效应蛋白);EPs)可能调节细胞外基质的重塑、抗病毒/细菌反应的激活以及肠道和外周的氧稳态等过程。免疫刺激蛋白CCL17、CXCL2、CXCL8(又称IL8)、IFNA、IFNL1、HMOX1、ITGB3和THBS1的基因表达在ETEC不存在或不存在的情况下均有所升高。当细胞单独暴露于ETEC时,没有观察到其中一些免疫刺激蛋白的表达升高。此外,所有提取物都高度刺激了线粒体/过氧化物酶体同体SLC25A21基因反义RNA在ec挑战细胞中的表达。这种反义RNA对SLC25A21翻译的抑制可能会在细胞质和细胞器内部造成2-氧二甲酸和2-氧戊二酸的浓度梯度,这两种物质都是脂肪酸β氧化的代谢物。结论:ETEC应激小肠上皮细胞暴露于微藻提取物影响了这些细胞的“脂肪酸β氧化”、ATP和活性氧的产生以及前胶原链中赖氨酸残基的(去)羟基化。特异性EPs和免疫刺激蛋白的基因表达升高表明,微藻提取物作为饲料/食品添加剂,可以引导人类和单胃动物肠道中一系列代谢和免疫过程,这些过程受到肠道细菌病原体的胁迫。
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