This study aimed to screen differentially expressed proteins (DEPs) in the tibial growth plate of thiram-induced tibial dyschondroplasia (TD) in Daheng broilers and to conduct bioinformatics analysis, providing potential targets for exploring the molecular mechanism of TD.One-day-old Daheng broilers were randomly divided into control group (NC) and TD model group (fed with 100 mg/kg thiram). Tibial cartilage growth plate tissues were collected at 17 days of age, and HE staining was performed to confirm pathological changes. Three biological replicates per group were subjected to Astral DIA label-free quantitative proteomic sequencing, and DEPs were screened with |log₂FC| ≥ 1 and P < 0.05. Principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), GO annotation, KEGG pathway enrichment analysis, GSEA, and protein-protein interaction (PPI) network were performed to screen core candidate proteins. HE staining showed that the TD group exhibited significantly thickened tibial growth plate, disorganized chondrocyte arrangement, and typical cartilage plug formation, indicating successful TD model establishment. A total of 7 561 proteins were identified by proteomics, with 7 300 quantifiable proteins. PCA and PLS-DA analysis showed good separation between the two groups. A total of 297 DEPs were screened, including 131 up-regulated and 166 down-regulated proteins. GO analysis indicated that DEPs were mainly enriched in biological processes such as negative regulation of ossification, regulation of Notch signaling pathway, reactive oxygen species metabolism, and regulation of mitochondrial membrane potential; cellular components were mainly localized in the extracellular matrix; molecular functions mainly involved methyltransferase activity and heparin binding. KEGG significantly enriched pathways included ECM-receptor interaction, glycosaminoglycan biosynthesis, and focal adhesion. GSEA further validated the enrichment of pathways such as phagosome and phosphatidylinositol signaling system. PPI network analysis screened out core node proteins including BIRC3, BCL2, MRPS14, ASPN, POSTN, COL5A1, and DAP3. This study revealed the differential protein expression profile of thiram-induced TD in Daheng broilers at the protein level. The screened candidate proteins may be closely related to abnormal cartilage development, providing new clues for in-depth understanding of the pathogenesis of TD.