Hulst Marcel, van der Weide Rommie, Hoekman Arjan, van Krimpen Marinus
Wageningen Livestock Research, De Elst 1, 6708 WD Wageningen, The Netherlands.
2Application Centre for Renewable Resources, Wageningen University & Research, Edelhertweg 1, 8219 PH Lelystad, The Netherlands.
Genes Nutr. 2019 Mar 19;14:8. doi: 10.1186/s12263-019-0632-z. eCollection 2019.
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 (ETEC) or under non-stressed conditions, were exposed to micro algae extracts and changes in gene expression were recorded.
IPEC-J2 cells were exposed for 2 and 6 h to extracts prepared from the biomass of the microalgae (C), (H), (S), or a mixture of S and (AM), in the absence and presence of ETEC. Gene expression in cells was measured using porcine "whole genome" microarrays.
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.
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.
微藻在全球范围内被视为动物和人类饮食中蛋白质的替代来源。微藻还产生一系列具有潜在有益和促进健康作用的生物活性物质。为了更好地理解微藻作为饲料添加剂时的作用模式,将产肠毒素大肠杆菌(ETEC)应激或非应激条件下的猪肠上皮细胞(IPEC-J2)暴露于微藻提取物中,并记录基因表达的变化。
在有无ETEC的情况下,将IPEC-J2细胞分别暴露于从微藻(C)、(H)、(S)的生物质或S与(AM)的混合物制备的提取物中2小时和6小时。使用猪“全基因组”微阵列测量细胞中的基因表达。
单独的微藻提取物增强了一组编码从细胞分泌的具有生物活性的蛋白质的基因的表达。这些分泌蛋白(以下称为效应蛋白;EPs)可能调节细胞外基质重塑、抗病毒/细菌反应激活以及肠道和外周的氧稳态等过程。在有无ETEC的情况下,对所有四种提取物均观察到免疫刺激蛋白CCL17、CXCL2、CXCL8(别名IL8)、IFNA、IFNL1、HMOX1、ITGB3和THBS1的基因表达升高。当细胞仅暴露于ETEC时这些免疫刺激蛋白中的几种未观察到表达升高。此外,所有提取物在ETEC攻击的细胞中高度刺激线粒体/过氧化物酶体转运体SLC25A21基因的反义RNA的表达。这种反义RNA对SLC25A21翻译的抑制可能会在这些细胞器的细胞质和内部之间形成脂肪酸β氧化的两种代谢物2-氧代己二酸和2-氧代戊二酸的浓度梯度。
ETEC应激的肠上皮细胞暴露于微藻提取物会影响这些细胞中“脂肪酸β氧化”、ATP和活性氧的产生以及原胶原链中赖氨酸残基的(去)羟基化。特定EPs和免疫刺激蛋白的基因表达升高表明,微藻提取物用作饲料/食品添加剂时,可以引导受肠道细菌病原体应激的人类和单胃动物肠道中的一系列代谢和免疫过程。