Insights into cellular metabolic pathways of the combined toxicity responses of Caco-2 cells exposed to deoxynivalenol, zearalenone and Aflatoxin B1

Insights into cellular metabolic pathways of the combined toxicity responses of Caco-2 cells exposed to deoxynivalenol, zearalenone and Aflatoxin B1

Metabolic profiling in Caco-2 cells was studied for the combined toxic effects of deoxynivalenol (DON), zear-alenone (ZEN), and Aflatoxin B-1 (AFB(1)) through untargeted GC-MS. The GC-MS spectra of Caco-2 cells treated with individual 6.7 mu M DON, 20 mu M ZEN, 20 mu M AFB(1) and the combined DON + AFB(1) (6.7 + 20 mu M) and DON + ZEN + AFB(1) (6.7 + 20 + 20 mu M) for 24 h were deconvoluted, aligned and identified with MS DIAL. The metabolic pathway analysis was analyzed with MetaMapp and visualized with CytoScape. Results show that the combined DON + AFB(1) and DON + ZEN + AFB(1) treatment has an obvious "synergistic effect". The apoptosisrelated gene mRNA test result indicates that the combined mycotoxins downregulate Bcl-2 gene and upregulate Bax, p53, caspase-3, caspase-8 and caspase-utilized 9 genes, more significantly than any individual mycotoxins group. The cellular metabolism illustrated that the combined mycotoxin groups, DON + ZEN + AFB(1) seriously effect glycine, serine and threonine metabolism, pyruvate metabolism, etc. while no metabolic disorders were presented in individual mycotoxin group. Our hypothesis was validated that the combined mycotoxins with low concentrations can have a synergistic effect in the metabolism, which may lead to cellular apoptosis or necrosis.

通过非靶向气相色谱 - 质谱法研究了Caco - 2细胞中的代谢谱，以了解脱氧雪腐镰刀菌烯醇（DON）、玉米赤霉烯酮（ZEN）和黄曲霉毒素B - 1（AFB₁）联合的毒性作用。用单独的6.7 μM DON、20 μM ZEN、20 μM AFB₁以及联合的DON + AFB₁（6.7 + 20 μM）和DON + ZEN + AFB₁（6.7 + 20 + 20 μM）处理24小时的Caco - 2细胞的气相色谱 - 质谱光谱，用MS DIAL进行了解卷积、对齐和鉴定。代谢途径分析用MetaMapp进行分析，并使用CytoScape进行可视化。结果表明，联合的DON + AFB₁和DON + ZEN + AFB₁处理具有明显的“协同效应”。凋亡相关基因mRNA测试结果表明，联合的霉菌毒素下调Bcl - 2基因，并上调Bax、p53、caspase - 3、caspase - 8和caspase - 9基因，比任何单一霉菌毒素组更为显著。细胞代谢表明，联合的霉菌毒素组DON + ZEN + AFB₁严重影响甘氨酸、丝氨酸和苏氨酸代谢、丙酮酸代谢等，而单一霉菌毒素组未出现代谢紊乱。我们的假设得到了验证，即低浓度的联合霉菌毒素在代谢中可具有协同效应，这可能导致细胞凋亡或坏死。