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<jats:title>Abstract</jats:title><jats:p>Vitamin D has been intensively studied for its association with human health, but the scope of such association and the causal role of vitamin D remain controversial. We aim to comprehensively investigate the links between vitamin D and human health through both epidemiological and Mendelian randomization (MR) analyses. We examined the epidemiological associations between serum 25‐hydroxyvitamin D (25(OH)D) concentration and 90 diseases/traits in 326,409 UK Biobank (UKBB) Europeans. The causal relations between 25(OH)D and 106 diseases/traits were investigated by performing MR analysis using genome‐wide significant 25(OH)D‐associated variants (<jats:italic>N</jats:italic> = 143) from the largest UKBB GWAS to date. In epidemiological analysis, we found 25(OH)D was associated with 45 diseases/traits across cardiovascular/metabolic diseases, psychiatric/neurological diseases, autoimmune/inflammatory diseases, cancer, musculoskeletal diseases, and quantitative traits. In MR‐analysis, we presented evidence suggesting potential causal role of 25(OH)D in increasing height (<jats:italic>β</jats:italic> = .064, 95% confidence interval [CI] = 0.019–0.11) and preventing the risk of ovarian cancer (odds ratio [OR] = 0.96, 95% CI = 0.93–0.99), multiple sclerosis (OR = 0.96, 95% CI = 0.94–0.98), leg fracture (OR = 0.60, 95% CI = 0.45–0.80) and femur fracture (OR = 0.53, 95% CI = 0.32–0.84). These findings confirmed associations of vitamin D with a broad spectrum of diseases/traits and supported the potential causal role of vitamin D in promoting health.</jats:p>