Explaining combinatorial effects of mycotoxins Deoxynivalenol and Zearalenone in mice with urinary metabolomic profiling

Jian Ji,Jiadi Sun,Ivana Blaženović,Pei Zhu,Yinzhi Zhang,Xiulan Sun,Morteza Gholami,Fangchao Cui,Jean Habimana

doi:10.1038/s41598-018-21555-y

Abstract

Urine metabolic profiling of mice was conducted utilizing gas chromatography-mass spectrometry (GC-MS) to investigate the combinatory effect of mycotoxins deoxynivalenol (DON) and zearalenone (ZEN) on the metabolism of the mice. Experiments were conducted by means of five-week-old mice which were individually exposed to 2 mg/kg DON, 20 mg/kg ZEN and the mixture of DON and ZEN (2 mg/kg and 20 mg/kg, respectively). The intragastric administration was applied for three weeks and urine samples were collected for metabolic analysis. Univariate and multivariate analysis were applied to data matrix processing along with respective pathway analysis by MetaMapp and CytoScape. The results showed that the combined DON and ZEN administration resulted in lower significant changes, compared to the individual mycotoxin treated groups verified by heatmap. Metabolic pathways network mapping indicated that the combined mycotoxins treated groups showed a little effect on the metabolites in most pathways, especially in glucose metabolism and its downstream amino acid metabolism. In glucose metabolism, the content of galactose, mannitol, galactonic acid, myo-inositol, tagatose was drastically down-regulated. Furthermore, the organic acids, pyruvate, and amino acids metabolism displayed the same phenomenon. In conclusion, the combined DON/ZEN administration might lead to an “antagonistic effect” in mice metabolism.

Highlights

Mycotoxins are secondary metabolites of fungi which are toxic to both human and animals[1]
Previous studies showed that DON could inhibit the protein, DNA, and RNA synthesis leading to cell apoptosis[8]
The current study increasingly addresses the combined effects of mycotoxins on the metabolism and metabolic pathways changes

Summary

Results

The metabolites selected for heatmap based on the principles of FDR corrected p value (p-value < 0.05) were analyzed by ANOVA test (Table S4) They were split into three groups: the first metabolites group was significant (FDR p-value < 0.05) compared to all mycotoxins treated groups; the second metabolites group was significant (FDR p-value < 0.05) only in individual DON and ZEN groups; the third group of metabolites was significant (FDR p-value < 0.05) only in individual DON group. All the significant metabolites of the individual mycotoxin-treated were put into pathway analysis on a web-based tool, MetaboAnalyst 3.0. In the individual ZEN-treated group, the metabolites with fold change values > 10, including gluconic acid, citric acid, tagatose, galactose, galacturonic acid, xylitol, galactonic acid, 4-hydroxyphenylpyruvic acid, were mainly incorporated in the galactose metabolism and TCA cycle. Fold change direction was calculated in MetaMapp based on two factors, the t-test p-value and fold change value

Discussion

Conclusion

Methods