From Intestinal Barrier to Inflammatory Response: Combined Impact of Deoxynivalenol and Heat Stress in Mice (Raw test data)

Published: 16 April 2025| Version 1 | DOI: 10.17632/ypx66tywt8.1
Contributors:
Han Shi, Zilong Sun

Description

This study aims to investigate the combined effects of deoxynivalenol and heat stress on intestinal damage in mice, a critical issue given the increasing prevalence of heat stress in agricultural environments. Our research demonstrates that heat stress exacerbates the harmful effects of deoxynivalenol on intestinal health, leading to significant alterations in gut morphology, microbiota composition, and inflammatory responses.

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(1) Seven-week-old healthy ICR mice of both sexes were randomly divided into four groups and exposed to different heat stress conditions for model establishment. Basic physiological indicators were recorded daily, and after 14 days, the mice were euthanized for intestinal tissue examination. Oxidative stress markers were measured to assess sex- and temperature-dependent responses to heat stress. (2) Based on preliminary screening, seven-week-old healthy male ICR mice were randomly assigned to four groups: control (C), heat stress (H), DON exposure (D), and combined stress (DH). Basic physiological parameters were recorded daily, and after 14 days, cecal contents and duodenal tissues were collected for analysis. (3) 16S rDNA sequencing was performed to analyze gut microbiota composition. Hematoxylin-eosin (HE) staining and transmission electron microscopy (TEM) were used to examine intestinal morphology, while periodic acid-Schiff (PAS) staining assessed goblet cell changes. The expression levels of intestinal barrier-related proteins (Claudin-1, Occludin, ZO-1), glucose transporters (SGLT1, GLUT2), inflammatory cytokines (IL-1β, IL-6, TNF-α), and key molecules in the TLR/NF-κB signaling pathway (TLR2, TLR4, IKKβ, NF-κB) were determined via qRT-PCR and Western blotting. Enzyme-linked immunosorbent assays (ELISA) quantified TLR2, TLR4, and inflammatory cytokines at the protein level, while immunofluorescence analysis visualized TLR2, TLR4, and Occludin distribution in the intestine. Oxidative stress markers (ROS, MDA) and antioxidant enzyme activities (SOD, GSH-Px, CAT) were measured using commercial assay kits. Finally, the correlation between intestinal damage indicators and gut microbial taxa was analyzed to elucidate the comprehensive mechanism of stress-induced intestinal injury.

Institutions

  • Shanxi Agricultural University

Categories

Veterinary Medicine, Veterinary Environmental Contaminant

Funders

  • the Earmarked Fund for Modern Agro-industry Technology Research System Program
    Grant ID: 2024CYJSTX12-10

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