Acrylamide, which is commonly used in various industries, may also form in food products cooked in high temperatures. Glycidamide, the ultimate genotoxic metabolite of acrylamide, is generated within cells through CYP4502E1-mediated epoxidation. Recent studies have shown that acrylamide and/or glycidamide may cause infertility by disrupting spermatogenesis, decreasing germ cell production and sperm fertilization ability due to their toxic effects on the male reproductive system. This study aimed to determine some direct effects of acrylamide and glycidamide on antioxidant defenses and on steroidogenic enzymes of Leydig and Sertoli cells. For this purpose, mouse Leydig and Sertoli cells were exposed to acrylamide (0.01 or 1mmol/L) or to glycidamide (0.001 or 0.5mmol/L) for 24h. Following the exposure, antioxidant enzyme activities (catalase, superoxide dismutase, glutathione peroxidase and -glutamyl transpeptidase), cellular antioxidant levels (glutathione) and steroidogenic enzyme activities (3-hydroxysteroid dehydrogenase and 17-hydroxysteroid dehydrogenase) were calculated. It was shown that acrylamide and glycidamide may cause inhibition of antioxidant and steroidogenic enzymes in Leydig and Sertoli cells. In conclusion, acrylamide and glycidamide may alter testicular function, thereby disrupting male reproduction.