Insights Into the Toxic Effects of 3‐Monochloropropane‐1,2‐Diol on Sertoli Cells via Tandem Mass Tagging Proteomics

ABSTRACT

Sertoli cells (SCs) play a vital role in the seminiferous tubules, as they are involved in maintaining the microenvironment, providing structural and nutritional support for germ cells, and modulating the process of spermatogenesis. 3‐Monochloropropane‐1,2‐diol (3‐MCPD), a common food‐processing contaminant, is known to impair male reproduction by interfering with SC function. However, the underlying toxicological mechanisms of 3‐MCPD in SCs have not been fully elucidated. In this study, primary rat SCs exposed to 3‐MCPD were analyzed using tandem mass tag (TMT)–based proteomics. A total of 285 differentially expressed proteins (DEPs) were identified, including 165 upregulated and 120 downregulated proteins. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis enrichment analyses revealed that 3‐MCPD exposure significantly affected pathways related to glutathione (GSH) metabolism, oxidative stress response, retinoic acid (RA) metabolism, and meiosis. Biochemical assays demonstrated that 3‐MCPD exposure profoundly reduced intracellular GSH reserves and increased reactive oxygen species (ROS) levels. RT‐qPCR validation confirmed the changes of key functional genes, including Crabp1 , Rdh10 , Rbp1 , ErbB3/4 , Hmox1 , Nqo1 , Star , and Gstm1 . Importantly, exogenous RA supplementation not only restored the impaired meiotic paracrine signaling molecules Nrg1/3 but also alleviated ROS accumulation and redox imbalance. These findings elucidate the comprehensive proteomic alterations in SCs induced by 3‐MCPD and reveal a novel functional crosstalk between RA signaling and oxidative balance, offering new insights into potential intervention strategies against reproductive toxicity.