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Effects of deoxynivalenol (DON) and its microbial biotransformation product deepoxy-deoxynivalenol (DOM-1) on a trout pig mouse and human cell line

机译:脱氧雪腐烯醇(DON)及其微生物转化产物深氧脱氧雪腐烯醇(DOM-1)对鳟鱼猪小鼠和人细胞系的影响

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摘要

Deoxynivalenol (DON), a trichothecene produced by various Fusarium species, is one of the most prevalent food- and feed-associated mycotoxins. The effects of DON and deepoxy-deoxynivalenol (DOM-1) were assessed in five different cell lines from different tissues and species starting from the first line of defense, the trout gill (RTgill-W1) and pig intestinal cells (IPEC-1 and IPEC-J2) over immune cells, as second line of defense (mouse macrophages RAW 264.7) to human liver cells (HepG2). Viability was assessed with a WST-1 assay, except for RTgill-W1, where a neutral red (NR) and sulforhodamine B (SRB) assay was performed. Additionally, more sensitive parameters, such as interleukin-, nitric oxide (NO)-, and albumin-release were determined. Viability was affected by DON at concentrations starting at 10 μmol/L (RTgill-W1), 0.9 μmol/L (IPEC-1), 3.5 μmol/L (IPEC-J2), and 0.9 μmol/L (HepG2), whereas DOM-1 did not have such an effect. Additionally, NO was decreased (0.84 μmol/L DON), whereas interleukin (IL)-6 was increased (0.42 μmol/L DON) in lipopolysaccharide (LPS)-stimulated DON-, but not DOM-1-treated RAW cells. Tumor necrosis factor (TNF)-α release, however, was not affected. Interestingly, albumin secretion of HepG2 cells was decreased by both DON and DOM-1 but at a much higher concentration for DOM-1 (228 versus 0.9 μmol/L for DON). 98.9% of DOM-1 was retrieved by liquid chromatography tandem mass spectrometry at the end of the experiment, proving its stability. In this study, IL-6 was the most sensitive parameter, followed by NO and albumin release and viability for HepG2 and IPEC-1.
机译:镰刀菌属各种物种产生的单端孢霉烯脱氧雪腐烯酚(DON)是与食物和饲料相关的最常见的真菌毒素之一。从第一道防线,鳟鱼ill(RTgill-W1)和猪肠道细胞(IPEC-1和DH5)开始,在来自不同组织和物种的五个不同细胞系中评估了DON和深氧-脱氧雪腐烯酚(DOM-1)的作用。 IPEC-J2)作为人类肝细胞(HepG2)的第二道防线(小鼠巨噬细胞RAW 264.7)。除RTgill-W1外,均使用WST-1分析法评估生存力,其中RTgill-W1进行中性红(NR)和磺基罗丹明B(SRB)分析。此外,确定了更敏感的参数,例如白介素,一氧化氮(NO)和白蛋白释放。 DON在10μmol/ L(RTgill-W1),0.9μmol/ L(IPEC-1),3.5μmol/ L(IPEC-J2)和0.9μmol/ L(HepG2)的浓度下开始影响生存力。 -1没有这样的作用。此外,在脂多糖(LPS)刺激的DON-中,NO降低(0.84μmol/ L DON),而白介素(IL)-6升高(0.42μmol/ L DON),但未用DOM-1处理的RAW细胞。然而,肿瘤坏死因子(TNF)-α的释放不受影响。有趣的是,DON和DOM-1均可降低HepG2细胞的白蛋白分泌,但DOM-1的浓度要高得多(DON的浓度为228比0.9μmol/ L)。实验结束时通过液相色谱串联质谱法回收了98.9%的DOM-1,证明了其稳定性。在这项研究中,IL-6是最敏感的参数,其次是NO和白蛋白释放以及HepG2和IPEC-1的生存力。

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