Regulation of retinal hemodynamics in diabetic rats by increased expression and action of endothelin-1

Purpose. To investigate the role of endogenous endothelin-1 (ET-1) expression and its interaction with the ET(A) receptor in the physiologic regulation of vascular tone as well as in the development of abnormal retinal hemodynamics in diabetes. Methods. Retinal blood flow, using digitized video fluorescein angiography recordings, was quantitated after intravitreous injections of ET-1; BQ-123, an ET(A) receptor antagonist; and phosphoramindon, an endothelin converting enzyme inhibitor in the eyes of diabetic and nondiabetic rats. A total of 154 rats were used for these experiments. Message levels of preproendothelin-1 (preproET-1) were measured from the retina of diabetic and nondiabetic rats using competitive polymerase chain reaction (PCR) techniques. Results. Retinal blood flow was reduced (33%, P <0.001) in diabetic rats compared to nondiabetic rats. BQ-123, an ET(A) receptor antagonist, but not saralasin, an angiotensin receptor antagonist, increased retinal blood flow in a dose-dependent manner in diabetic (EC(50) of 8x10(-7) M) and in nondiabetic rats (EC(50) of 8x10(-8) M). Besides being resistant to BQ-123, the maximal response in diabetic animals occurred 20 minutes later than in nondiabetic animals. Decreasing ET-1 levels by inhibiting endothelin-converting enzyme with phosphoramidon normalized retinal blood flow in diabetic rats, In nondiabetic rats, the intravitreous injection of exogenous ET-1 (10(-8) M) resulted in retinal blood flow decreases comparable to those measured in diabetic animals, and the subsequent injection of 10(-4) M BQ-123 produced retinal blood flow changes comparable to those measured in BQ-123 injected diabetic rats, Comparison of preproET-1 messenger RNA expression in the retina, brain, and lung of control and diabetic rats using quantitative PCR and Northern blot analysis showed 2.0- and 1.7-fold increases in the retina and the brain, respectively, without changes in the lung. Conclusions. These data suggest that ET-1 is involved in the regulation of retinal blood flow in normal physiologic outcome, and an increase in the endogenous expression of ET-1 contributes to the reduction of retinal blood flow reported in the early stages of diabetes mellitus.