Drug exposure is a common risk factor of male infertility and should be investigated carefully. This study aimed to evaluate the possible protective effect of α-lipoic acid against gentamicin induced-testicular damage. Sixty Wister male rats aged 8 weeks and weighted 200±20 gm were randomly assigned to six groups of ten. Groups 1 and 2 were treated with 18.25 and 36.5mg/kg gentamicin, respectively. Groups 3, 4, and 5 were treated with 36.5mg/kg gentamicin plus 100mg/Kg/day α-lipoic acid (ALA), 200mg/Kg/day ALA, and 100mg/kg vitamin-E, respectively. Group 6 served as control. All treatments were administered once daily for 14days. Half the animals in each group was sacrificed on the 15th day ,while the second half was sacrificed on the 60th day of the experiment. Blood samples were collected from the retro-orbital venous plexus just before sacrifice. Immediately after the sacrifice, reproductive organs were removed and weighed, and semen was collected for analysis. Groups treated with gentamicin and either ALA or vitamin-E showed preserved reproductive organs weight and sperm parameters compared to gentamicin alone on both sacrifice days (p<0.05). Testosterone level was also preserved with either ALA or vitamin-E co-administrated with gentamicin compared to gentamicin alone (p<0.05). However, gonadotropin levels showed variable levels with the cotreatments compared to gentamicin alone. The hormone levels of all groups were approximately normalized on the second sacrifice day. Pathological lesions induced by gentamicin were markedly reduced upon cotreatment with ALA. Adding α-lipoic acid or vitamin-E to gentamicin therapy protects from gentamicin-induced reproductive system-impairment in experimental animals.