This study investigated the relationship between vitamin D levels and male infertility in 306 men with suspected infertility. Participants were divided into two groups based on vitamin D levels (<20 ng/mL and ≥20 ng/mL). The study found that higher vitamin D levels were significantly correlated with improved sperm motility (types A and B), while lower levels were associated with reduced motility (types C and D). Vitamin D was positively correlated with testosterone levels but not with other hormones or essential minerals like iron, calcium, magnesium, or phosphorus. Functional enrichment analysis suggested that vitamin D may influence male infertility through pathways related to infection, immune response, and signaling molecules such as HIF-1 and FOXO. These findings indicate that adequate vitamin D levels may contribute to better reproductive health by improving sperm motility and hormone levels. The study highlights the potential role of vitamin D in male infertility and suggests that further research is needed to fully understand its mechanisms.This study investigated the relationship between vitamin D levels and male infertility in 306 men with suspected infertility. Participants were divided into two groups based on vitamin D levels (<20 ng/mL and ≥20 ng/mL). The study found that higher vitamin D levels were significantly correlated with improved sperm motility (types A and B), while lower levels were associated with reduced motility (types C and D). Vitamin D was positively correlated with testosterone levels but not with other hormones or essential minerals like iron, calcium, magnesium, or phosphorus. Functional enrichment analysis suggested that vitamin D may influence male infertility through pathways related to infection, immune response, and signaling molecules such as HIF-1 and FOXO. These findings indicate that adequate vitamin D levels may contribute to better reproductive health by improving sperm motility and hormone levels. The study highlights the potential role of vitamin D in male infertility and suggests that further research is needed to fully understand its mechanisms.