In this study, the harmful effects of doxorubicin on rat testicular tissue and the protective role of thymoquinone and resveratrol were investigated. The study employes 10 groups, each consisting of 8 male Wistar albino rats, for a total of 80 male rats. The groups were set up as follows: control (C), doxorubicin (D), resveratrol (R) (5 mg·kg−1, 20 mg·kg−1), thymoquinone (T) (5 mg·kg−1, 20 mg·kg−1), and combination groups (D + R 5 mg·kg−1, D + R 20 mg·kg−1, D + T 5 mg·kg−1, D + T 20 mg·kg−1). The C group received physiological saline by oral gavage every other day. Groups administered D at a dose of 15 mg·kg−1 intraperitoneally on day 10. Groups receiving T and R were given every other day for 21 days. At the end of the study period, the animals were euthanized by cervical dislocation under anesthesia. Testis and epididymis samples were collected with attention to sterility for spermatological, histopathological, and immunohistochemical analyses. The findings showed that sperm concentration decreased in all groups treated with D, motility and mitochondrial activity decreased, and acrosome and membrane integrity were impaired. Histopathological and immunohistochemical data also revealed significant decreases in germinal cell layer thickness, tubule diameter, Johnson's testicular score, and relative testicular weight, while the B-cell lymphoma 2 (Bcl-2) ratio decreased and Bcl–2–associated X protein (Bax) expression increased (P<0.05). Additionally, the T 5 mg·kg−1 group exhibited an adverse effecton sperm density, motility, membrane integrity, and mitochondrial activity. The T 20 mg·kg−1, R 5 mg·kg−1, and R 20 mg·kg−1 groups, on the other hand, yielded more positive results than the control group for many parameters. When considering the combined groups, the D + R 5 mg·kg−1 and especially the D + R 20 mg·kg−1 groups successfully prevented the toxicity caused by the D group in terms of both spermatological and histopathological and immunohistochemical parameters. In conclusion, R was found to have a stronger protection against D-induced testicular toxicity compared to thymoquinone.