The Mechanism of Zinc Sulfate in Improving Fertility in Obese Rats Analyzed by Sperm Proteomic Analysis

This study investigates the mechanism underlying the improving effect of zinc on fertility in obese rats using proteomics. The effects of three different doses of ZnSO₄ on spermatogenesis and hormone levels were studied. Testicular spermatogenesis was observed by HE staining. Serum estrogen and testosterone levels were measured by chemiluminescent microparticle immunoassay. Sperm proteomic analysis was performed by liquid chromatography-mass spectrometry. The DAVID database was used to perform the GO enrichment analysis and KEGG pathway analysis of the differentially expressed genes, and the STRING online database was used to construct a PPI network. The sperm count, sperm motility, and testosterone hormones of the ZnSO₄-treated rats group were increased. ZnSO₄ improved testicular structure and spermatogenesis abnormalities caused by obesity. Proteomic analysis showed that there were 401 differentially expressed proteins in a total of 6 sperm samples from the ZnSO₄-treated group and the obesity groups. Differential proteins were input into the DAVID website. The 341 identified proteins were then classified according to their biological functions. The KEGG analysis showed that the enriched signal pathways included glycolysis/gluconeogenesis, carbon metabolism, citrate cycle, fatty acid metabolism, and pyruvate metabolism. Some proteins were shown to be associated with valine, leucine, and isoleucine degradation pathways. STRING analysis obtained 36 node proteins. Cytoscape analysis showed that these proteins mainly participated in nine networks including metabolic process, oxidation-reduction, aerobic respiration, RNA splicing, and glutathione conjugation. ZnSO₄ may improve the fertility of obese male rats by regulating protein expression related to metabolism, inflammation, and sperm maturation.

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