Research on male sterility mechanism of Cucurbita pepo L. is of great value for enhancing breeding and hybrids production. Using male sterile （X-MS） and fertile plants （X-MF） of zucchini as experimental materials， this study integrated transcriptome sequencing with analysis of endogenous hormone contents， antioxidant enzyme activities， sucrose/starch content. Results demonstrated that during anther developmental stages-tetrad （T1）， early uninucleate （T2）， mid-late uninucleate （T3）， binucleate （T4）， pre-anthesis （T5）， the abscisic acid content， gibberellin content， and auxin content （T1-T3） of X-MS male flower buds lower than X-MF， exhibited reduced performance， an increasing trend in peroxidase activity， and decreasing trend in starch and sucrose content. Transcriptome sequencing identified 1947 differentially expressed genes （DEGs） from T1 to T3， comprising 911 up-regulated and 1036 down-regulated genes. GO enrichment analysis revealed catalytic activity and binding as the most annotations in molecular function， while cellular and metabolic processes dominated biological process annotations. KEGG analysis showed significant DEGs enrichment in pathways such as plant hormone signal transduction， phenylpropanoid biosynthesis， starch and sucrose metabolism， and amino sugar and nucleotide sugar metabolism. Further analysis of plant hormone signaling， phenylpropanoid biosynthesis， and starch/sucrose metabolism pathways identified candidate genes including one peroxidase-related gene， one sucrose synthase-related gene， and two IAA-related genes related to. These genes may play pivotal roles in pollen development of male-sterile zucchini and provide mechanistic insights for future research on male sterility in zucchini.