Abstract: Background and Objective: Spermatogenesis is a process by which germ cells produce spermatozoa and can be disturbed at every level. Nitric Oxide Synthases (NOS), implicate in interactions with Oxidative Stress (OS) which is one of the main factors in the etiology of male infertility. The High Fat Diet (HFD) is a major factor of obesity which in turn is important for enhancing OS. Antioxidants and garlic could attenuate or reverse effects of HFD. The aim of the study was to investigate the effects of dietary antioxidants and garlic on testicular inducible NOS (iNOS) and endothelial NOS (eNOS) in Wistar albino rats fed on HFD. Materials and Methods: Groups (each n = 8) were: SD (100% access to standard diet), F-HFD, (100% access to HFD) and R-HFD (70% access to HFD), F-HFD +antioxidants, F-HFD+garlic and R-HFD+antioxidants. The HFD consisted of a 60% fatty diet in 3 forms: Without antioxidants, with antioxidants and with garlic. The testicular iNOS and eNOS were studied by immunohistochemical (IHC) method. Also used ANOVA, repeated measures ANOVA, t-tests and Tukey’s test (where necessary) to analyze the data (p<0.05). Results: The iNOS increased in the F-HFD and R-HFD+antioxidants groups. The eNOS increased in R-HFD,F-HFD and F-HFD+garlic groups. The H-E evaluation in R-HFD group showed a decrease in spermatogenesis score count and seminiferous tubules diameters (μm) in comparison with the SD and F-HFD groups. R-HFD+antioxidants group had lower score than F-HFD+antioxidants and F-HFD+garlic groups. Conclusion: Restricted fat diet consumption causes increase in weight and impairs spermatogenesis. Results of this study reveal that adding the antioxidants can’t improve histological changes of testis. The iNOS expression in seminiferous tubules in restricted fat diet along with antioxidants, suggest a potential role of iNOS in spermatogenesis and male infertility.