ObjectiveTo investigate the relationship between single nucleotide polymorphisms (SNPs) related to vitamin D (VitD) metabolism and post-stroke depression (PSD) in patients with ischemic stroke.MethodsA total of 210 patients with ischemic stroke were enrolled at the Department of Neurology in Xiangya Hospital, Central South University, from July 2019 to August 2021. SNPs in the VitD metabolic pathway (VDR,CYP2R1,CYP24A1, andCYP27B1) were genotyped using the SNPscan™multiplex SNP typing kit. Demographic and clinical data were collected using a standardized questionnaire. Multiple genetic models including dominant, recessive, and over-dominant models were utilized to analyze the associations between SNPs and PSD.ResultsIn the dominant, recessive, and over-dominant models, no significant association was observed between the selected SNPs in theCYP24A1andCYP2R1genes and PSD. However, univariate and multivariate logistic regression analysis revealed that theCYP27B1rs10877012 G/G genotype was associated with a decreased risk of PSD (OR: 0.41, 95% CI: 0.18–0.92,p= 0.030 and OR: 0.42, 95% CI: 0.18–0.98,p= 0.040, respectively). Furthermore, haplotype association analysis indicated that rs11568820-rs1544410-rs2228570-rs7975232-rs731236 CCGAA haplotype in theVDRgene was associated with a reduced risk of PSD (OR: 0.14, 95% CI: 0.03–0.65,p= 0.010), whereas no significant association was observed between haplotypes in theCYP2R1andCYP24A1genes and PSD.ConclusionOur findings suggest that the polymorphisms of VitD metabolic pathway genesVDRandCYP27B1may be associated with PSD in patients with ischemic stroke.