Objective To investigate pathological damage and functions after subchronic exposure to sodium fluoride (NaF) on the heart, liver, kidneys, and testes of male rats, and to explore the possible mechanism. Methods Thirty-two Wistar male rats were randomly divided into control, and low, medium, and high dose groups with eight rats in each group. NaF at doses of 0, 12, 24, and 48 mg / kg body weight was applied by intragastric administration once a day at 6 days per week and for 16 weeks. The occurrence of dental fluorosis and body mass in each group of rats was recorded, and the organ coefficients of the heart, liver, kidneys, and testes of rats were calculated. Serum levels of SOD, CAT, GSH-Px, and MDA were measured by ELISA. Pathological changes of the heart, liver, kidneys, and testes were observed by HE staining. Biochemical method were used to determine the myocardial enzyme profile, liver functions, and kidney functions in rats, and analyze the sperm quality of rats. Results Compared with rats in the control group, rats in low, medium, and high dose groups showed different degrees of fluorosis changes in maxillary and mandibular incisors, indicating successful modeling. The body weight of rats in the high dose group was lower than that in the other three groups (P<0. 05). The renal organ coefficient in low and high dose groups was higher than that in the control group (P<0. 05). Compared with that in the control group, CAT and GSH-Px activity was decreased in medium and high dose groups (P<0. 05). The serum level of MDA in in the high dose group was increased (P<0. 05). Compared with the findings in the control group, HE staining showed different degrees of pathological damage in the heart, liver, kidneys, and testes of low, medium, and high dose groups. Compared with those in the control group, LDH and CK-MB levels were increased in each dose group (P<0. 05). The serum levels of ALT and AST were increased in medium and high dose groups (P<0. 05), and the serum level of GGT was increased in the high dose group (P<0. 05). Serum levels of BUN and Scr were increased in low, medium, and high dose groups (P<0. 05). The sperm count of rats was decreased in low, medium, and high dose groups, and sperm motility and the sperm malformation rate were increased ( P< 0. 05). Conclusions Subchronic exposure to sodium fluoride damages multiple organs in rats, and the mechanism may be related to oxidative stress induction.