Background: Alzheimer’s disease (AD) is a common disease in the elderly that is associated with impaired metabolism and biology of the hippocampus. Although the role of exercise and natural antioxidants in improving the disease has been reported, the role of muscle contraction-related physical activity along with royal jelly (RJ) consumption is not yet well understood. Objectives: This study aimed to investigate the effect of eight weeks of training on positive slope (ETPS) and negative slope (ETNS) with royal jelly (RJ) consumption on O-6-methylguanine DNA methyltransferase (MGMT) and ATPase gene expression levels in the hippocampus tissue of trimethyltin (TMT)-induced AD rats. Methods: In this experimental trial, 36 male Sprague-Dawley AD rats (induced with 8 mg/kg TMT) were divided into (1) control + normal saline/royal jelly solvent) (Sh); (2) ETPS; (3) ETNS; (4) ETPS + RJ; (5) ETNS + RJ, and (6) RJ groups. Six rats were placed in the healthy control (HC) group. Then the training groups trained on ( + 15 and -15) slopes for eight weeks, five sessions per week, and each session lasted 60 minutes at a speed of 16 m/min. The royal jelly (RJ) groups received 100 mg/kg royal jelly per day by intraperitoneal injection. Results: ETNS, ETPS, ETPS + RJ, and ETNS + RJ increased MGMT gene expression (P ≤ 0.05). ETPS and ETPS + RJ also increased ATPase gene expression (P ≤ 0.05). However, RJ had no significant effect on increasing MGMT and ATPase gene expression in the hippocampus tissue of AD rats (P ≥ 0.05). Conclusions: It seems that the improvement of DNA damage markers and energy levels depends on the type of contraction. Although training on positive and negative slopes with royal jelly consumption has an interactive effect on DNA repair markers, training on a positive slope and royal jelly consumption has an interactive effect on ATPase gene expression.