Evaluation of Metabolic Characteristics Induced by Deoxynivalenol in 3D4/21 Cells

Abstract

Deoxynivalenol (DON) is a common mycotoxin that causes immunosuppression in pigs. Its effects on cellular metabolism remain unclear. In this study, we investigate DON-induced metabolic alterations in porcine alveolar macrophage cell line 3D4/21 using non-targeted metabolomics. MTT assays showed DON reduced cell viability in a concentration- and time-dependent manner. Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) revealed distinct metabolic profiles between control and DON-treated groups. Metabolomic analysis identified 127 differential metabolites (VIP > 1, p < 0.05), primarily in purine metabolism, glutathione metabolism, and arginine-proline metabolism. Integration with transcriptomic data confirmed that these pathways play key roles in DON-induced immunotoxicity. Specifically, changes in purine metabolism suggested disrupted nucleotide synthesis and energy balance, while glutathione depletion indicated weakened antioxidant defense. These findings provided a systems biology perspective on DON's metabolic reprogramming of immune cells and identified potential therapeutic targets to reduce mycotoxin-related immunosuppression in swine.

Keywords: alveolar macrophages; deoxynivalenol; metabolomics; pig.

Figure 1

Effects of different concentrations of DON (0, 1, and 2 μM) on the viability of 3D4/21 cells assessed by MTT assay at 12 h (a), 24 h (b), and 48 h (c) after treatment. Data are presented as mean ± SD (n = 8). Different letters indicate significant differences among groups (p < 0.05, one-way ANOVA followed by Tukey’s multiple comparisons).

Figure 2

Principal component analysis score plot of the control and DON groups. Cells were treated with 2 μM DON for 24 h. (a) PCA score plot for the two groups analyzed in the positive ion mode. (b) PCA score pt for the two groups analyzed in the negative ion mode. PC1 was the first principal component; PC2 was the second principal component.

Figure 3

Orthogonal partial least squares discriminant analysis (OPLS-DA) score and permutation test plot in the positive ion mode (a,c) and the negative ion mode (b,d) for the control and DON groups. Cells were treated with 2 μM DON for 24 h. The intercept limit of Q² was calculated by the regression line, which was the plot of Q2 from the permutation test in the OPLS-DA model.

Figure 4

KEGG pathway enrichment analysis (a) and KEGG classification (b) of differential metabolites in 3D4/21 cells exposed to 2 µM DON for 24 h.

Figure 5

Integrated KEGG pathway enrichment analysis of metabolomic and transcriptomic data in 3D4/21 cells exposed to 2 µM DON for 24 h. Meta represents metabolomic data, and Tra represents transcriptomic data.