# Integrated Targeted and Untargeted Metabolomics Reveals the Toxic Mechanisms of Zearalenone in Goat Leydig Cells

## TL;DR

This study shows how zearalenone, a mycotoxin, harms male goat reproductive cells by disrupting hormone production and metabolism.

## Contribution

The study reveals ZEA's toxic effects on male ruminant Leydig cells through integrated metabolomics and cytotoxicity analysis.

## Key findings

- ZEA reduces Leydig cell viability and induces apoptosis and cell cycle arrest.
- ZEA disrupts steroid hormone synthesis and alters pathways like glycerophospholipid and choline metabolism.
- 52 altered metabolites were identified, linked to reproductive impairment in male goats.

## Abstract

This study focuses on the toxic mechanisms of zearalenone (ZEA) in goat Leydig cells (LCs). Using various experimental techniques, it is found that ZEA damages LCs in a dose-dependent manner, induces cell apoptosis and cycle arrest, and disrupts steroid hormone synthesis, as well as metabolic pathways such as glycerophospholipid and choline metabolism. This study clarifies the multifaceted hazards of ZEA to male goat reproductive cells and provides theoretical support for evaluating the impact of ZEA on goat reproductive performance.

Zearalenone (ZEA) is a mycotoxin commonly found in animal feed and is associated with pronounced reproductive toxicity. However, most studies on ZEA’s reproductive effects have focused on female monogastric animals, while research on male ruminants remains limited. This study aimed to investigate the cytotoxic and metabolic mechanisms underlying ZEA-induced damage in goat Leydig cells (LCs). The CCK8 assay was first used to determine the effective ZEA concentration (IC50 ≈ 20 μM), and a cytotoxicity model was subsequently established. The model’s validity was confirmed using qRT-PCR, transmission electron microscopy, flow cytometry, and JC-1 staining. Results showed that ZEA significantly reduced LCs viability in a dose-dependent manner, decreased mitochondrial membrane potential, induced cell cycle arrest, and triggered apoptosis. Targeted and untargeted metabolomics analyses revealed that ZEA disrupts steroidogenic pathways and alters steroid hormone secretion, resulting in elevated levels of progesterone, corticosterone, and androstenedione, and reduced dihydrotestosterone levels. Furthermore, 52 significantly altered metabolites were identified, predominantly enriched in glycerophospholipid metabolism, choline metabolism, and neurotransmitter vesicle pathways, with corresponding changes in gene expression. Collectively, this study has confirmed that ZEA causes harm to the reproductive cells of male goats in multiple aspects, underscoring the link between metabolic dysregulation and reproductive impairment, and offering a foundation for evaluating ZEA’s impact on goat reproductive performance.