Ischemia/reperfusion is one of the emergency cases that frequently occurs in testis. This pathologic event is one of the reasons for infertility in men. Inflammation and oxidative stress induce ischemia/reperfusion injury in testis. Consequently, agents possessing antioxidant activity are applied in the treatment of testicular ischemia/reperfusion. In the present study, the effect of berberine administration in the treatment of testicular ischemia/reperfusion injury is investigated. In this experiment, 24 Wistar rats were randomly divided into four groups (n=6): Sham group (receiving normal saline 0.9%), control group (ischemia/reperfusion), treatment I group (ischemia/reperfusion group receiving 50 mg/kg berberine), and treatment II group (ischemia/reperfusion receiving 100 mg/kg berberine). All injections were performed through the intraperitoneal route. Histopathological findings demonstrated that in the Sham group, testis has normal structure and normal spermatogenesis occurs. In the ischemia group, severe hyperemia, coagulative necrosis, and interstitial edema are observed and spermatogenesis has severe damage. In treatment I group (50 mg/kg of berberine), moderate interstitial edema, hyperemia, and coagulative necrosis are observed. Besides, spermatogenesis has moderate damage. In group treated with a high dose of berberine (100 mg/kg), all damages are mild. This experiment reveals that berberine exerts its protective impact in a dose-dependent manner so that the highest protective impact is observed in the group treated with 100 mg/kg of berberine. With respect to the major role of testicular ischemia in infertility and the results of the present study, berberine can be used as a valuable plant extract in the treatment of testicular ischemia and preventing its harmful impacts.