To investigate the effects of zinc supplementation for male rats fed with high fat diet on their circRNA and microRNA (miRNA) expression profiles in the testicular tissue. Methods: The rat models with high-fat diet and zinc supplementation were established in two groups, and the testicular tissues of the rats in the two groups were selected for the high-throughput RNA sequencing, and the differentially expressions of circRNAs and miRNAs were screened. GO and KEGG functional enrichment analysis was performed, and then the circRNA,-miRNA, and-mRNA network was constructed. Results: There were 54 differentially expressed circRNAs screened and 2 differentially expressed miRNAs screened. The results of GO analysis showed that the fatty acid oxidation, the lipoprotein metabolism and biosynthesis, the peptide transport ATPase activity, and the oxidoreductase activity played their roles. KEGG enrichment analysis showed that the signaling pathways of the TNF, the cAMP, and the AMPK had been enriched. The circRNA and miRNA with significant differences and with targeting relationship were screened, which including novel_circ_0016964, novel_circ_0013783, and rno-miR-540-3p, and then the target gene of rno-miR-540-3p was predicted, and the ceRNA network of circRNA, miRNA, and mRNA was constructed. Conclusion: Zinc supplementation for the rats fed with high-fat diet can change the expression profiles of their circRNA and miRNA during their reproductive injury, so circRNA and miRNA may be as the potential targets for playing their roles.