This work was aimed to observe the protective role of β-carotene (β-CAR) against renal toxicity induced by gibberellic acid (GA3), a plant growth hormone commonly used in adult male rats. The studied rats were divided into six groups of eight each: group one served as a control, while Groups 2 and 3 received GA3 at concentrations of 50 ppm and 75 ppm in drinking water, respectively; Group 4 was orally administered β-carotene at the amount of 50 mg; Groups 5 and 6 were treated by both GA3 and β-CAR. All treatments were made daily for 90 days. Results revealed that exposure to GA3 significantly increased serum urea and creatinine with a concomitant reduction in serum albumin and total serum protein, indicating kidney dysfunction. Furthermore, GA3 significantly raised the values of ROS and LPO in the kidneys but lowered the activities of GPx, CAT, and SOD. These biochemical alterations were further confirmed by comet assay procedures representing photomicrographs of DNA damage and tail migration in kidney tissues. Histopathological examination showed marked pathological changes in the kidneys of all the GA3-treated groups. Importantly, co-administration with β-CAR reduced these toxic effects, restoring biochemical parameters to near normal compared to GA3-intoxicated rats. Histopathological findings further revealed reduced damage to kidneys in groups treated with both GA3 and β-CAR. This study therefore concludes that β-CAR has substantial kidney protective potential against GA3-induced kidney toxicity in rats.