This study was conducted to investigate the effects of ACTH application on kidney function tests, electrolytes, and hematological parameters in rats. In the research two groups were formed, including control and stress. Each group consisted of 6 male Sprague Dawley rats which were about 13 weeks old and 200-250 g in weight. Serum isotonic was administered at a dose of 0.5 ml/rat to control group, and ACTH was applied at a dose of 4 mg/kg to stress group at once intraperitoneally. Blood samples were taken 3 hours after the application. Concentrations of glucose, amylase, creatine, creatine kinase (CK), creatine kinase MB (CK-MB), urea, uric acid, lactate dehydrogenase (LDH), calcium (Ca), phosphorus (P), magnesium (Mg) sodium (Na), chlorine (Cl) and potassium (K) in serum; the numbers of total leukocytes (WBC), platelet (PLT) and erythrocytes (RBC) and the values of hemoglobin (HGB) and hematocrit (HCT), on whole blood were performed. The level of glucose, WBC, monocyte %, granulocyte %, p <= 0.001 values of stress group were significantly higher than those of control group (p <= 0.001) while the values of CK, CK-MB, urea, BUN were significantly higher (p <= 0.05) while lymphocytes % (p <= 0.001) and the levels of Na, Cl, and K (p <= 0.05) were significantly lower in stress group compared to control. These results indicated that stress induced by ACTH had noticeable impact on the renal function test, electrolyte balance and hematological parameters.