INTRODUCTION: The pathophysiology of subfertility in men with varicocele remains unclear as well as the role of inhibin B (Inh-B) and anti-Müllerian hormone (AMH). The aim of this study was to evaluate Inh-B and AMH concentrations in the spermatic vein of subfertile men with varicocele.PATIENTS AND METHODS: A total of 61 subfertile men with varicocele and 31 fertile controls underwent standard andrological evaluation. All subfertile men underwent varicocelectomy, during which blood samples were obtained from the spermatic vein to evaluate Inh-B and AMH concentrations.RESULTS: Peripheral vein Inh-B concentrations in men with varicocele were lower as compared to controls (52.9 [8.3-136.0) vs 116±9.7 ng/dL, P = .001). There was no difference in AMH concentrations (10.2 [4.4-45.4]) vs 10.4±0.8 pg/dL, P = 0.9). Spermatic vein Inh-B concentrations in men with varicocele were higher compared to those of peripheral vein (87.6±4.4 vs 52.9 [8.3-136.0] ng/dL, P = .001). On the contrary, spermatic vein AMH concentrations were lower compared to those from peripheral vein (8.84 [3.9-47.7] vs 10.2 [4.4-45.4] pg/dL, P = .013).CONCLUSIONS: Inh-B constitutes a reliable marker of Sertoli cell function as well as spermatogenesis. On the contrary, the clinical significance of AMH in men with varicocele remains to be elucidated.