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Deoxynivalenol (DON) is a type of mycotoxin that threatens human and livestock health. Right open reading frame kinase 2 (RIOK2) is a kinase that has a pivotal function in ribosome maturation and cell cycle progression. This study aims to clarify the role of the RIOK2 gene in DON-induced cytotoxicity regulation in porcine intestinal epithelial cells (IPEC-J2). Cell viability assay and flow cytometry showed that the knockdown of RIOK2 inhibited proliferation and induced apoptosis, cell cycle arrest, and oxidative stress in DON-induced IPEC-J2. Then, transcriptome profiling identified candidate genes and pathways that closely interacted with both DON cytotoxicity regulation and RIOK2 expression. Furthermore, RIOK2 interference promoted the activation of the MAPK signaling pathway by increasing the phosphorylation of ERK and JNK. Additionally, we performed the dual-luciferase reporter and ChIP assays to elucidate that the expression of RIOK2 was influenced by the binding of transcription factor Sp1 with the promoter region. Briefly, the reduced expression of the RIOK2 gene exacerbates the cytotoxic effects induced by DON in IPEC-J2. Our findings provide insights into the control strategies for DON contamination by identifying functional genes and effective molecular markers. [ABSTRACT FROM AUTHOR]

Deoxynivalenol (DON), as a secondary metabolite of fungi, is continually detected in livestock feed and has a high risk to animals and humans. Moreover, pigs are very sensitive to DON. Recently, the role of histone modification has drawn people's attention; however, few studies have elucidated how histone modification participates in the cytotoxicity or genotoxicity induced by mycotoxins. In this study, we used intestinal porcine epithelial cells (IPEC-J2 cells) as a model to DON exposure in vitro. Mixed lineage leukemia 1 (MLL1) regulates gene expression by exerting the role of methyltransferase. Our studies demonstrated that H3K4me3 enrichment was enhanced and MLL1 was highly upregulated upon 1 μg/mL DON exposure in IPEC-J2 cells. We found that the silencing of MLL1 resulted in increasing the apoptosis rate, arresting the cell cycle, and activating the mitogen-activated protein kinases (MAPKs) pathway. An RNA-sequencing analysis proved that differentially expressed genes (DEGs) were enriched in the cell cycle, apoptosis, and tumor necrosis factor (TNF) signaling pathway between the knockdown of MLL1 and negative control groups, which were associated with cytotoxicity induced by DON. In summary, these current results might provide new insight into how MLL1 regulates cytotoxic effects induced by DON via an epigenetic mechanism. [ABSTRACT FROM AUTHOR]

Deoxynivalenol (DON) caused serious cytotoxicity for animal cells. However, genes involved in regulating DON toxicity and the underlying molecular mechanisms remain largely unknown. This study explored the role of SLC4A11 and MFSD3 in alleviating DON toxicity and analyzed the DNA methylation changes of these two genes. Viability and cell cycle analysis showed that DON exposure decreased the IPEC-J2 viability (P < 0.01), blocked the cell cycle in the G2/M phase (P < 0.01), and increased the rate of apoptosis (P < 0.05). Expression of the SLC4A11 and MFSD3 genes was significantly downregulated upon DON exposure (P < 0.01). Overexpression of SLC4A11 and MFSD3 can enhance the cell viability (P < 0.01). DNA methylation assays indicated that promoter methylation of SLC4A11 (mC-1 and mC-23) and MFSD3 (mC-1 and mC-12) were significantly higher compared with those in the controls and correlated negatively with mRNA expression (P < 0.05). Further analysis showed that mC-1 of SLC4A11 and MFSD3 was located in transcription factor binding sites for NF-1 and Sp1. Our findings revealed the novel biological functions of porcine SLC4A11 and MFSD3 genes in regulating the cytotoxic effects induced by DON, and may contribute to the detection of biomarkers and drug targets for predicting and eliminating the potential toxicity of DON. [ABSTRACT FROM AUTHOR]

Deoxynivalenol (DON) is a trichothecenes produced by fungi that is widespread and poses a threat to human and animal health. The Notch1 signalling pathway is tightly involved in cell fate determination. The aim of this study was to investigate the role of the Notch1 signalling pathway in DON exposure. Herein, we found that the Notch1 signalling pathway was significantly activated after DON exposure, while Notch1 expression was negatively regulated by DON-induced ROS. Then, the Notch1 signalling pathway was blocked by the γ-secretase inhibitor DAPT in DON exposure. Flow cytometry analysis and antioxidant parameter measurements revealed that DAPT treatment significantly aggravated the oxidative stress induced by DON. The detection of apoptosis showed that DAPT treatment increased the cell apoptotic rate. Further analysis revealed that inhibiting the Notch1 signalling pathway reduced autophagy upon DON exposure. RT–qPCR and Western blot analysis showed that inhibiting the Notch1 signalling pathway aggravated cellular inflammation and activated the MAPK pathway, indicating that the MAPK pathway may be the downstream signalling pathway. Taken together, our research revealed that the Notch1 signalling pathway is essential for protection against DON. Inhibition of Notch1 signalling increases oxidative stress, causes cell apoptosis, reduces autophagy and aggravates cell inflammation after DON exposure. This study investigated the role of the Notch1 signalling pathway in DON exposure and provided a basis for exploring the mechanism of DON. [Display omitted] • The Notch1 signalling pathway is activated after DON exposure. • Notch1 signalling pathway is essential for antioxidant against DON. • Notch1 signalling pathway participates in apoptosis and autophagy induced by DON. • MAPK pathway is activated after inhibiting Notch1 signalling pathway upon DON exposure. [ABSTRACT FROM AUTHOR]

Deoxynivalenol (DON) is one of the mycotoxins that is toxic to agricultural environment, which poses high risks to human and farm animal health. Noncoding RNAs have been shown to be crucial regulators of toxicological processes and as promising biomarkers for toxicity monitoring and prevention of mycotoxin contamination. Herein, we characterized genome-wide transcriptional profiling of porcine intestinal epithelial cells upon DON exposure and illustrated a subset of miRNAs and lncRNAs involved in the cellular processes by targeting genes associated with stress responses. A total of 110 differential expression miRNAs and 143 differential expression lncRNAs were identified between the DON exposure and control cell samples. Interactive network analysis showed that miR-330 was one hub noncoding RNA, expression of which was significantly increased upon DON exposure. Functional enrichment analysis indicated that the genes involved in the networks were mainly enriched in the terms of plasma membrane bounded cell projection assembly, mRNA processing, and regulation of mitochondrion organization. Further functional analysis revealed that high expression of miR-330 inhibits the reactive oxygen species production, cell apoptosis, and autophagic flux in cells upon DON exposure. Luciferase assay further indicated that miR-330 could directly target MAPK15. Knockdown of MAPK15 resulted in decreased reactive oxygen species level and cell apoptosis induced by DON, indicating the existence of miR-330-MAPK15 regulatory axis in regulating DON toxicity. Our work shed novel insights into the mode of action of DON at cellular level and indicated the potential of miR-330 as a biomarker for toxicity monitoring of DON contamination, which contributes to the development of effective biomonitoring and prevention strategies to reduce the toxicological effects of DON. [Display omitted] • DON-associated transcriptomic alterations and regulatory networks were explored. • miR-330 regulated DON toxicity and showed potential as a biomarker for DON exposure. • miR-330 can directly target the MAPK15 gene. • miR-330-MAPK15 regulatory axis was involved in DON-induced toxicological processes. [ABSTRACT FROM AUTHOR]

Deoxynivalenol (DON) is a common environmental toxin that is secreted by fusarium fungi that frequently contaminates feedstuff and food. While the detrimental effects of DON on human and animal reproductive systems have been well recognized, the underlying mechanism remains poorly understood. Ovarian granulosa cells (GCs), which surround oocytes, are crucial for regulating oocyte development, mainly through the secretion of hormones such as estrogen and progesterone. Using an in vitro model of murine GCs, we characterized the cytotoxic effects of DON and profiled genome-wide chromatin accessibility and transcriptomic alterations after DON exposure. Our results suggest that DON can induce decreased viability and growth, increased apoptosis rate, and disrupted hormone secretion. In total, 2533 differentially accessible loci and 2675 differentially expressed genes were identified that were associated with Hippo, Wnt, steroid biosynthesis, sulfur metabolism, and inflammation-related pathways. DON-induced genes usually have a concurrently increased occupancy of active histone modifications H3K4me3 and H3K27ac in their promoters. Integrative analyses identified 35 putative directly affected genes including Adrb2 and Fshr, which are key regulators of follicular growth, and revealed that regions with increased chromatin accessibility are enriched with the binding motifs for NR5A1 and NR5A2, which are important for GCs. Moreover, DON-induced inflammatory response is due to the activation of the NF-κB and MAPK signaling pathways. Overall, our results provide novel insights into the regulatory elements, genes, and key pathways underlying the response of ovarian GCs to DON cytotoxicity. [ABSTRACT FROM AUTHOR]

Melatonin is an important endogenous hormone that shows antioxidant functions and pleiotropic effects, playing a crucial role in animal reproduction. Ovary granulosa cells (GCs) surround the oocyte, which play an important role in regulating oocytes development. Deoxynivalenol (DON) is a common fusarium mycotoxin contaminant of feedstuff and food, posing a serious threat to human and animal reproductive systems. Herein, murine ovary GCs were studied as a reproduction cell model, aimed to assess the protective effect of melatonin on DON-induced toxicity in murine ovary GCs. The results showed that DON adversely affected the viability and growth of murine ovary GCs and increased the apoptosis rate, while melatonin administration ameliorated these toxic effects. We further reveal that DON exposure increased the intracellular reactive oxygen species level, reduced the mitochondrial membrane potential and ATP, and upregulated Tnfα (tumor necrosis factor α), Il6 (interleukin 6), and Il1β (interleukin 1 β) gene expression. Moreover, DON exposure downregulated reproductive hormone gene expression and significantly increased nuclear factor kappa B (p65) activation and mitogen-activated protein kinase phosphorylation. Melatonin treatment attenuated all these effects, suggesting that melatonin protects GCs from the adverse effects of DON by ameliorating oxidative stress, mitochondrial dysfunction, and inflammation. Overall, these results reveal the mechanisms of DON and melatonin in GCs and provide a theoretical basis for melatonin as a drug to improve mycotoxin contamination. [ABSTRACT FROM AUTHOR]

Deoxynivalenol (DON) is widely found in food and feed, posing a threat to human and animal health. Lycopene (Lyc) is a natural plant extracts with significant antioxidant properties. This study was conducted to investigate the protective effects of Lyc on IPEC-J2 cells upon DON exposure. The detection of cell viability and trypan blue staining showed that Lyc alleviated cell damage and decreased cell apoptotic rate induced by DON. The analysis of reactive oxygen species (ROS) level and antioxidant parameter measurements showed that Lyc significantly down-regulated the content of ROS and restored antioxidant enzyme activity. Furthermore, mitochondrial membrane potential (ΔΨm) detection, mitochondrial DNA copy number (mtDNAcn) assay and adenosine triphosphate (ATP) concentration detection showed Lyc improved mitochondrial function after DON exposure. The results of transcriptome analysis, ROS detection and CCK8 assay suggested that Lyc may activated the oxidative phosphorylation (OXPHOS) to improve mitochondrial function. Conclusively, our results suggested that Lyc alleviated DON-induced oxidative stress by improving mitochondrial function through OXPHOS signaling pathway. [Display omitted] [ABSTRACT FROM AUTHOR]

This study explored protective effects of melatonin (MEL) on deoxynivalenol (DON)-induced toxicity in porcine jejunum epithelial cells (IPEC-J2). Cells were preexposed to MEL and then exposed to DON to detect cell viability, apoptosis, and oxidative stress indicators. Compared to DON treatment, pretreatment with MEL significantly increased cell proliferation. (P < 0.01), intracellular catalase (CAT) and superoxide dismutase (SOD) levels (P < 0.05), decreased apoptosis and oxidative stress, and significantly attenuated the inflammatory response. RNA-seq analysis revealed that MEL protects IPEC-J2 from the adverse effects of DON by affecting the expression of tight junction and autophagy pathway-related genes. Similarly, further experiments revealed that MEL partly prevented DON-induced disruption of intestinal barrier function and decreased autophagy induced by DON via activation of the AKT/mTOR pathway. In conclusion, these results demonstrated the preventive properties of MEL against DON-induced cell damage by activating the antioxidant system and Inhibition of autophagy. • This study firstly descripted the underlying molecular mechanisms of (Melatonin) MEL ameliorated the Deoxynivalenol (DON)-induced toxicity in porcine jejunum epithelial cells (IPEC-J2). • MEL partly prevented DON-induced disruption of intestinal barrier function. • MEL decreased the autophagy induced by DON via activating the AKT/mTOR pathway. [ABSTRACT FROM AUTHOR]