In the current study, the effects of enhancing semen quality, brain neurotransmitters, and antioxidant status in rabbits under heat stress with acacia gum, vitamin C, and lycopene as dietary supplements were estimated in vitro and in silico. A total of 40 males from New Zealand White rabbits aged 3months were equally divided into four groups (n¼10). The first group was a control. The other three groups were the heat stress group (oral acacia gum, 100mg/kg body weight), the ascorbic acid group (30 mg/kg body weight), and the lycopene group (50 mg/kg body weight). All semen showed physical characteristics in terms of increasing sperm motility, motility index, sperm normality, and live sperm as compared to the control group. On the other hand, sperm abnormalities, and dead sperm significantly (p<0.001) decreased compared with the control group. It is of interest to show that most semen traits were significantly (p<0.001) better for acacia gum than vitamin C and lycopene. The expression of neurotransmitters (5-HT, DA, Glu, Asp) was enhanced in all treatments compared to the control. The results of acacia gum were better (p<0.001) than those obtained with vitamin C and lycopene. The acacia gum treatment had a lower plasma semen MDA and NO (p<0.001) and higher GSH, TAC, SOD, CAT, L-Car, Naþ-Kþ ATPase, ATP, and total calcium content (p<0.001) than in other treatments and controls. These results were confirmed by the prediction of the binding energy, several conventional and carbon H-bonds, hydrophobicity, and SAS through the in silico docking analysis results. It can be concluded that acacia gum, vitamin c, and lycopene are used for enhancing semen quality, brain neurotransmitters, and the antioxidant status of rabbits under heat stress