This study aimed to evaluate the antioxidant activity of skim milk (SM) and permeate (P) in vitro and to evaluate their protective role against nicotine (NT)-induce oxidative stress, disturbances in sex hormones and histological changes in different organs of female rats. Four groups of female rats were treated orally for 4 weeks included the control group, NT-treated group (0.6 mg/kg b.w), NT plus SM-treated group (100 mg/kg b.w) and NT plus P-treated group (200 mg/kg b.w). Blood and tissue samples were collected at the end of the experiment for different analyses. The in vitro results showed that SM and P have a diphenyl 1-2 pieryl- hydrazil (DPPH) scavenging activity in a dose-dependent and SM was more effective than P. The in vivo results showed that NT administration induced a significant reduction in superoxide dismutase (SOD), reduced glutathione (GSH), follicle-stimulating hormone (FSH) and luteinizing hormone (LH) accompanied by significant histological changes in ovary, colon, and urinary bladder. Co-administration of NT plus SM or P could alleviate the histological changes, improved the antioxidants enzyme activity, and normalized FSH and LH. The overall results showed that SM was more effective than P. It could be concluded that these milk by-products can be used as a dietary supplement to reduce the risk of tobacco smoking or tobacco chewing.