Abstract Objective The present study was carried out to investigate the possible protective role of copper (II) albumin against zinc oxide nanoparticles (ZnONPs) provoked DNA damage and hepatotoxicity in rats. Methods Forty adult male Sprague Dawley rats were randomly divided into fve groups; Group I: the control group maintained on a regular diet, Group II: received 1 ml/day of milk as the solvent of Cu (II) albumin complex, Group III: received Cu (II) albumin at 0.03 µg/gm, Group IV: exposed to ZnONPs (400 mg/kg/day), and Group V: exposed to 400 mg/kg/day ZnONPs plus Cu (II) albumin. All treatments were administered for 30 days. At the end of the experiment, animals were euthanized for collection of blood and liver samples. DNA damage in blood and liver was evaluated by using comet assay, while hepatotoxicity was evaluated from histopathological changes of hepatic tissue and liver enzymes (ALT and AST). The oxidative status parameters including nitric oxide (NO), glutathione peroxidase, malondialdehyde (MDA), and total antioxidant capacity (TAC) were also measured. Results The results showed that ZnONPs induced oxidative stress through a signifcant increase in MDA and NO activities, a signifcant decrease in TAC, and slight decrease in glutathione peroxidase. Signifcant DNA damage, a signifcant increase in AST, and a slight increase in ALT were accompanied by histological changes in the liver ZnONPs exposed group. Concurrent Cu (II) albumin supplement to ZnONPs-treated rats in Group IV reversed most of the histopathological changes and DNA damage, signifcantly lowered ALT and AST levels as well as MDA and NO, and elevated the TAC and GPx. Conclusion Based on these results, it can be concluded that Cu (II) albumin efectively protects against ZnONPs-induced hepatic dysfunction and DNA damage in rats.