Background Nicotine administration can generate severe oxidative stress and lipid peroxidation. Royal jelly, with its antioxidant properties, acts as a scavenger of reactive oxygen species. This study describes the apitherapy effects of royal jelly on testicular damage following nicotine administration. Methods Forty‐eight male BALB/c mice were divided into 8 groups ( n = 6): saline, 3 different doses of royal jelly (100, 150, and 200 mg/kg body weight (BW) per day), nicotine (1.5 mg/kg), and 3 different groups of Nic + Roy (1.5 mg/kg of Nic + 100, 150, and 200 mg/kg BW per day of royal jelly). Nicotine was administrated intraperitoneally, and royal jelly was prescribed orally for 10 consecutive days. Serum levels of hormones (testosterone, luteinizing hormone, and follicle‐stimulating hormone), total antioxidant capacity, nitric oxide (NO) status, malondialdehyde levels, sperm DNA fragmentation, sperm parameters, histopathological changes (H&E staining), immunohistochemistry against apoptotic proteins, and gene expression of Bcl ‐ 2 , p53 , Caspase ‐ 3 , and Nrf2 (real‐time PCR) were assessed to evaluate the molecular and histological changes. Results Hormone levels, sperm parameters, and status of antioxidants were decreased significantly ( p < .05) following nicotine administration. Moreover, royal jelly treatment normalized hormonal and antioxidant characteristics, decreased apoptotic gene expression, increased Nfr2 gene expression, and restored histopathological alteration to the physiological status significantly ( p < .05). Conclusion Royal jelly upregulates the antioxidant status, inhibits the mitochondrial‐dependent apoptosis pathway, and increases the rate of proliferation. This therapeutic agent effectively protected the testis against nicotine‐associated damages by antioxidant and anti‐apoptotic effects.