ABSTRACT Background: Vitamin D has multiple biological effects on male reproductive system. Vitamin D deficiency (VD−) and disturbance of lipid metabolism can induce changes in testicular hormone production and seminal parameters that relate to male infertility.   Objective: To investigate the role of vitamin D and lipid metabolism on fertility in male albino rats. Methods: The study was performed on 60 male rats, divided into 6 groups; GI: control group, GII: orlistat group, GIII: orlistat and vitamin D group, GIV: hyperlipidemic group, GV: hyperlipidemic orlistat group and GVI: hyperlipidemic orlistat and vitamin D group. Serum total cholesterol (TC), triglycerides (TG), low density lipoprotein cholesterol (LDL-c) and high density lipoprotein cholesterol (HDL-c), 25-hydroxyvitamin D (25(OHD), testosterone (T), inhibin B, follicle-stimulating hormone (FSH) and estradiol (E) levels were determined. Sperm count and viability were also analyzed. Results: Administration of orlistat in GII caused significant alterations of the serum lipid profile, compared to control group. There was interaction between dietary fat and VD− on serum 25 (OHD). A significant VD− was occurred in both GII and GIV, compared to GI. VD− in these groups consequently caused significant decrease in serum T, inhibin B, sperm count and viability and significant increase in FSH and E levels. Administration of orlistat in GV caused significant changes in serum lipid profile, a decrease in E, increase in 25(OHD) and inhibin B, compared to IV. On the other hand, administration of vitamin D with orlistat in GIII and GVI caused significant increase in HDL-c, 25(OHD), T, sperm count and viability and significant decrease in TC, TG, LDL-c, FSH and E. Conclusion: Both hyperlipidemia and hypolipidemia were associated with vitamin D deficiency. Moreover, vitamin D has a positive potential effect on male fertility in either hyperlipidemic or hypolipidemic rats.