title: Increased nitric oxide activity compensates for increased oxidative stress to maintain endothelial function in rat aorta in early type 1 diabetes.
abstract: Hyperglycaemia and oxidative stress are known to acutely cause endothelial dysfunction in vitro, but in the initial stages of diabetes, endothelium-dependent relaxation is preserved. The aim of this study was to investigate how endothelium-dependent relaxation is maintained in the early stages of type 1 diabetes. Diabetes was induced in Sprague-Dawley rats with a single injection of streptozotocin (48 mg/kg, i.v.), and after 6 weeks, endothelium-dependent and endothelium-independent relaxations were examined in the thoracic aorta in vitro. Lucigenin-enhanced chemiluminescence was used to measure superoxide generation from the aorta. Diabetes increased superoxide generation by the aorta (2,180 ± 363 vs 986 ± 163 AU/mg dry tissue weight). Acetylcholine (ACh)-induced relaxation was similar in aortae from control (pEC(50) 7.36 ± 0.09, R                 (max) 95 ± 3 %) and diabetic rats (pEC(50) 7.33 ± 0.10, R                 (max) 88 ± 5 %). The ACh-induced relaxation was abolished by the combined presence of the nitric oxide synthase inhibitor N-nitro-L-arginine (L-NNA, 100 μM) and an inhibitor of soluble guanylate cyclase, 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ, 10 μM) in control rats, but under the same conditions, the diabetic aortic rings showed significant relaxation to ACh (pEC(50) 6.75 ± 0.15, R                 (max) 25 ± 4 %, p < 0.05). In diabetic aortae, the addition of haemoglobin, which inactivates nitric oxide, to L-NNA + ODQ abolished the response to ACh. The addition of the potassium channel blockers, apamin and TRAM-34, to L-NNA + ODQ also abolished the relaxation response to ACh. Diabetes significantly elevated plasma total nitrite/nitrate and increased expression of endothelial nitric oxide synthase (eNOS) and calmodulin in aortae. These data indicate that after 6 weeks of diabetes, despite increased oxidant stress, endothelium-dependent relaxation is maintained due to the increased eNOS expression resulting in increased NO synthesis. In diabetic arteries, NO acts both through and independently of cGMP pathways to cause relaxation.
authors.affiliation: Department of Pharmacology, University of Melbourne, Melbourne, Victoria, Australia. anjali.jo@gmail.com
authors.forename: A
authors.initials: A
authors.lastname: Joshi
authors.forename: O L
authors.initials: OL
authors.lastname: Woodman
mesh_terms: D000109:Acetylcholine / Q000494:pharmacology; D000818:Animals; D001013:Aorta, Thoracic / Q000187:drug effects / Q000378:metabolism; D006152:Cyclic GMP / Q000378:metabolism; D003921:Diabetes Mellitus, Experimental / Q000503:physiopathology*; D003922:Diabetes Mellitus, Type 1 / Q000503:physiopathology*; D004730:Endothelium, Vascular / Q000378:metabolism; D008163:Luminescent Measurements; D008297:Male; D009569:Nitric Oxide / Q000096:biosynthesis / Q000378:metabolism*; D052250:Nitric Oxide Synthase Type III / Q000235:genetics; D018384:Oxidative Stress; D051381:Rats; D017207:Rats, Sprague-Dawley; D013481:Superoxides / Q000378:metabolism; D014664:Vasodilation / Q000187:drug effects; D014665:Vasodilator Agents / Q000494:pharmacology
chemical_list: D014665:Vasodilator Agents; D013481:Superoxides; D009569:Nitric Oxide; D052250:Nitric Oxide Synthase Type III; D006152:Cyclic GMP; D000109:Acetylcholine