We aimed to compare the preventive effects of combined antioxidants (CA1, 2) with a single antioxidant drug (thymoquinone; TQ) on experimental testis Ischemia/Reperfusion (I/R) injury. Thirty-five adult male Wistar rats were divided into five groups of seven rats each: control, testis I/R, testis I/R+CA1, testis I/R+CA2, and testis I/R+TQ. After 1h of testicular ischemia, reperfusion was achieved by detorsion for 4h. Antioxidants were intraperitoneally administered for 30min prior to reperfusion. All rats were sacrificed 4h after reperfusion to evaluate the tissue levels of malondialdehyde (MDA) and total antioxidant status (TAS) and the immunohistochemical evaluation of tissue inducible and endothelial nitric acid synthase (iNOS, eNOS) and apoptosis protease-activating factor 1 (APAF-1). MDA levels were lower and TAS values were higher in the I/R+antioxidant groups than in the I/R group (p<0.05). iNOS and eNOS levels in the I/R+antioxidant groups were also lower than those in the I/R group (p<0.05). There were no significant differences between the CA groups and the TQ group according to aforementioned parameters. In addition, tissue APAF-1 values were significantly higher in the I/R group than in the other groups. However, there was a significant difference between the TQ and CA groups in APAF-1 levels, which were highest in the TQ group (p<0.05). Although TQ alone increased TAS values and reduced tissue iNOS and eNOS levels, combined antioxidant treatment may more effectively reduce apoptosis and increase preventive effects in testis I/R injury.