BACKGROUND Carbohydrate restriction in diet is becoming a popular means of losing weight nowadays, although it has been reported that excessive intake of low-carbohydrate and high-protein (LCHP) diet causes an adverse effect on cardiovascular function. OBJECTIVE To investigate the influence of LCHP on erectile function in rats. METHODS A total of 48, 12-week-old rats were divided into 2 groups and either fed a LCHP diet (LCHP group) or a normal diet (Control group). Hematological examination, blood pressure evaluation, erectile function assessments as well as evaluations of the relaxation and contractile responses of corpus cavernosum were carried out in these rats by using standardized methods. Statistical analysis using 2-way ANOVA and Welch's t-test was conducted to examine the obtained data. METHODS At the end of the study period, the evaluated outcomes to assess erectile function were intracavernosal pressure , mean arterial pressure , endothelial functions, nitric oxide (NO)-operated nerve functions and the expressions of endothelial nitric oxide synthase (eNOS), neuronal nitric oxide synthase (nNOS), and sphingosine-1-phosphate receptor 1 (S1P1). RESULTS The intracavernosal pressure / mean arterial pressure ratio was significantly lower in the LCHP group (P < .05) at 4 weeks. Compared to the Control group, the LCHP group exhibited significantly lower responses to ACh and EFS and a decreased nNOS mRNA expression. The results based on this animal model indicate that extreme carbohydrate restricted diet may affect erectile function. Our study identified that LCHP decreased erectile function in rats. A major limitation of this study is, due to the extreme condition of completely replacing carbohydrates with protein, that carbohydrate intake will be gradually increased in the future. CONCLUSIONS Extreme carbohydrate restriction and high protein in diet may cause ED with vascular endothelial dysfunction and a decrease in the relaxation response of the corpus cavernosum smooth muscle via NO-operated nerves. Kataoka T, Hidaka J, Suzuki J, et al. Evaluating the Effects of Low Carbohydrate and High Protein Diet on Erectile Function in Rats. Sex Med 2021;10:100500.