Protective Role of Hydrogen Gas on Oxidative Damage and Apoptosis in Intestinal Porcine Epithelial Cells (IPEC-J2) Induced by Deoxynivalenol: A Preliminary Study.

Xu Ji,Wen Yao,Weijiang Zheng

doi:10.3390/toxins12010005

Abstract

To explore the protective role of hydrogen gas (H2) on oxidative damage and apoptosis in intestinal porcine epithelial cells (IPEC-J2) induced by deoxynivalenol (DON), cells were assigned to four treatment groups, including control, 5 μM DON, H2-saturated medium, and 5 μM DON + H2-saturated medium treatments. After 12 h of different treatments, the cell viability, biomarkers of cell redox states, and gene expression of antioxidant enzymes and apoptosis were observed and detected. Furthermore, caspase-3 and Bax protein expressions were measured by Western blot analysis. Our results demonstrated that the 5 μM DON significantly caused cytotoxicity to IPEC-J2 cells by reducing cell viability and increasing lactate dehydrogenase release in culture supernatants. Moreover, DON treatments significantly increased levels of 8-hydroxy-2′-deoxyguanosine, 3-nitrotyrosine, and malonaldehyde; however, they decreased total superoxide dismutase and catalase activities and downregulated messenger RNA (mRNA) expression related to antioxidant enzymes in cells. The 5 μM DON treatment also downregulated Bcl-2 expression and upregulated caspase-3 and Bax expression. However, the H2-saturated medium significantly improved cell growth status and reversed the change of redox states and expression of genes and proteins related to apoptosis induced by DON in IPEC-J2 cells. In conclusion, H2 could protect IPEC-J2 cells from DON-induced oxidative damage and apoptosis in vitro.

Highlights

Mycotoxins are toxic secondary metabolites from fungi which often exist as contaminants in animal and human food worldwide
Key Contribution: In this study, for the first time, we described that H2 precisely exhibited protective effects against oxidative damage and apoptosis of IPEC-J2 cells induced by DON in vitro
To observe the cytotoxic effects of DON on the growth of IPEC-J2 cells, we firstly evaluated cell viability using the Methyl Thiazolyl Tetrazolium (MTT) assay

Summary

Introduction

Mycotoxins are toxic secondary metabolites from fungi which often exist as contaminants in animal and human food worldwide. The consumption of mycotoxin-contaminated feed and food is considered a major health risk to animals and humans [1]. Investigated the contamination of DON in foodstuffs from different provinces in China between. They found that the occurrence rate of DON was over 74.5%, in which the average concentration ranged from 450.0–4381.5 μg/kg, suggesting that DON was a prevalent contaminant in China [4]. Following ingestion of a DON-contaminated diet, a reduction in growth and immunomodulating properties is induced [5]. DON can induce inflammation and oxidative stress, thereby accelerating cell apoptosis and influencing intestinal epithelial cell growth and function [7,8,9]

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Conclusion