The role of the sodium hydrogen exchanger-1 in mediating diabetes-induced changes in the retina.
BACKGROUND
The sodium hydrogen exchanger (NHE) is a transmembrane protein responsible for alkalinization and control of intracellular acidosis by the removal of hydrogen and the subsequent influx of sodium. Our investigation attempts to determine the role of NHE-1 in the pathogenesis of early retinal microangiopathy due to diabetes.


METHODS
Diabetes was induced in male Sprague-Dawley rats with a single intravenous streptozotocin injection (65 mg/kg). To examine the duration-dependent changes in NHE-1 expression, retinas from 1-, 6- and 12-week diabetic animals were analyzed. To examine the functional consequences of NHE-1 inhibition in comparison with good blood glucose control, diabetic rats were randomly assigned to poorly controlled diabetic, well-controlled diabetic, poorly controlled diabetic with cariporide groups and were compared with nondiabetic controls after six weeks. Cariporide is an orally active inhibitor of NHE-1 (6000 ppm in rat chow). At the end of the treatment period, color Doppler ultrasound was used to determine the resistivity index (RI) of the central retinal artery. The mRNA expression of endothelin (ET) isoforms 1 and 3, inducible and endothelial nitric oxide synthase (iNOS and eNOS respectively) and NHE-1 were examined. NHE-1 distribution was localized with immunohistochemistry.


RESULTS
All diabetic animals showed hyperglycemia, increased glycated hemoglobin and lower body weight gain compared to nondiabetic controls. Diabetes caused an increased RI, indicative of retinal vasoconstriction, which was corrected by both cariporide treatment and good glucose control. NHE-1 was localized in the endothelium of the retinal microvasculature and the neuronal and glial components. NHE-1 mRNA expression was unchanged after 1 week and increased after 6 and 12 weeks of diabetes. Furthermore, a diabetes-induced upregulation of ET-1 and ET-3 mRNA expression after six weeks was corrected with cariporide treatment. NHE-1 inhibition of diabetic animals upregulated iNOS mRNA levels, although expression of eNOS and iNOS mRNA were not altered in poorly controlled diabetes. Improved blood glucose control with higher doses of insulin also corrected diabetes-induced increased RI by upregulating eNOS and iNOS mRNA expression.


CONCLUSIONS
The results of the study suggest that NHE-1 may be involved in the regulation of several vasoactive modulators that contribute to functional alterations in diabetic retinal microangiopathy.
Cukiernik
Mark
M
University of Western Ontario, Department of Pathology, Canada.
Hileeto
Denise
D
Downey
Donal
D
Evans
Terry
T
Khan
Zia Ali
ZA
Karmazyn
Morris
M
Chakrabarti
Subrata
S
D000818:Animals; D003921:Diabetes Mellitus, Experimental / Q000473:pathology / Q000503:physiopathology*; D003930:Diabetic Retinopathy / Q000473:pathology / Q000503:physiopathology*; D019332:Endothelin-1 / Q000235:genetics; D019334:Endothelin-3 / Q000235:genetics; D005786:Gene Expression Regulation / Q000502:physiology*; D006146:Guanidines / Q000494:pharmacology; D008297:Male; D019001:Nitric Oxide Synthase / Q000235:genetics; D052250:Nitric Oxide Synthase Type III; D051381:Rats; D017207:Rats, Sprague-Dawley; D012160:Retina / Q000503:physiopathology*; D017923:Sodium-Hydrogen Exchangers / Q000037:antagonists & inhibitors / Q000235:genetics*; D013450:Sulfones / Q000494:pharmacology; D013997:Time Factors
D019332:Endothelin-1; D019334:Endothelin-3; D006146:Guanidines; D017923:Sodium-Hydrogen Exchangers; D013450:Sulfones; C069139:growth factor-activatable Na-H exchanger NHE-1; C093373:cariporide; D019001:Nitric Oxide Synthase; D052250:Nitric Oxide Synthase Type III; C496316:Nos3 protein, rat