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Aim: To analyze RhoA expression and activation in the aorta of diabetic rats. Methods: Male SD rats (n=70) were divided into 2 groups: the diabetic group and the control group. Diabetes was induced by intravenous injection of streptozotocin (55 mg/kg). The Rats were studied 3 weeks after the induction of diabetes. Western blotting was used to measure the expression and activation of Rho. Results: Heart rate was measured 24 h/d; it decreased by 58±13 beats/min in the diabetic rats. Isometric tension showed that the contraction of diabetic aorta was significantly reduced compared with that of control aorta when stimulated by KCl and serotonin. The relaxation of the diabetic aorta was reduced when stimulated by acetylcholine. An enhanced RhoA translocation in the aortic tissues of diabetic rats was determined by a 90% increase in membrane-bound RhoA, indicating that the activation of RhoA is markedly increased in the diabetic aorta. Conclusion: Our data suggest that upregulated RhoA could be involved in the vascular dysfunction of diabetic rats.