The Effects of in vitro Hyperglycemic Incubation on A23187-Mediated Contractile Responses of Rat Thoracic Aorta

  • Işıl Özakca Ankara University Faculty of Pharmacy
Keywords: Diabetes mellitus, Vascular smooth muscle, Endothelium, Hyperglycemia, Aging


Objective: Vascular tonus has been controlled by several factors secreted from endothelium in physiological conditions. In the present study, the possible changes on endothelium-derived contractile responses in normoglycemic and hyperglycemic conditions in terms of exposure-time dependency has been investigated. Methods: To assess the possible alterations under acute hyperglycemia with different incubation periods on endothelium-derived contractile responses in isolated rat thoracic aorta, A23187-mediated contractile responses were performed in a cumulative manner for isometric tension measurements. Results: Incubation for 3 hours with Krebs solution containing high glucose increased the A23187-mediated contraction of rat thoracic aorta. The A23187-induced contraction significantly decreased in response to the same incubation period with normoglycemic conditions and totally abolished with incubation for 6 hours. The possible effects of osmotic pressure induced by high glucose content checked with mannitol. Conclusion: Our results indicated that the A23187-mediated contractile response was increased by acute hyperglycemia. These data has shown the detrimental effects of short term hyperglycemia on endothelium-derived contractile factors. Also the incubation period-related characteristics of A23187-mediated contractile responses as observed in normoglycemic conditions indicate the unstable properties of endothelium-derived contractile factors and/or related signaling pathway(s).


Giacco F, Brownlee M. Oxidative Stress and Diabetic Complications. Circ Res 2010; 107: 1058-70.

Brownlee M. The Pathobiology of Diabetic Complications: a Unifying Mechanism. Diabetes 2005; 54: 1615-25.

Feletou M, Vanhoutte PM. Endothelial dysfunction: a multifaceted disorder. Am J Physiol Heart Circ Physiol 2006; 291: H985-1002.

Matsumoto T, Kobayashi T, Kamata K. A therapeutic target for microvascular complications in diabetes: endothelium-derived hyperpolarizing factor. Curr Cardiol Rev 2006; 2: 185-91.

Boulanger CM, Morrison KJ, Vanhoutte PM. Mediation by M3-muscarinic receptors of both endothelium-dependent contraction and relaxation to acetylcholine in the aorta of the spontaneously hypertensive rat. Br J Pharmacol. 1994; 112:519-24.

Koga T, Takata Y, Kobayashi K, Takishita S, Yamashita Y, Fujishima M. Age and hypertension promote endothelium-dependent contractions to acetylcholine in the aorta of the rat. Hypertension 1989; 14: 542-54.

Katusic ZS, Shepherd JT, Vanhoutte PM. Endothelium-dependent contractions to calcium ionophore A23187, arachidonic acid, and acetylcholine in canine basilar arteries. Stroke 1988; 19: 476-9.

Tang EH, Leung FP, Huang Y, Félétou M, So KF, Man RY et al. Calcium and reactive oxygen species increase in endothelial cells in response to releasers of endothelium-derived contracting factor. Br J Pharmacol 2007; 151: 15-23.

Vanhoutte PM, Tang EHC. Endothelium-dependent contractions: when a good guy turns bad!. J Physiol. 2008; 586: 5295-304.

Ellinsworth DC, Shukla N, Fleming I, Jeremy JY. Interactions between thromboxane A2, thromboxane/prostaglandin (TP) receptors and endothelium-derived hyperpolarization. Cardiovasc Res 2014; 102: 9-16.

Vanhoutte PM, Feletou M, Taddei S. Endothelium-dependent conractions in hypertension. Br J Pharmacol 2005; 144: 449-58.

Qu C, Leung SWS, Vanhoutte PM, Man RYK. Chronic Inhibition of Nitric-Oxide Synthase Potentiates Endothelium-Dependent Contractions in the Rat Aorta by Augmenting the Expression of Cyclooxygenase-2. J Pharmacol Exp Ther 2010; 334: 373-80.

Feletou M, Verbeuren Tj, Vanhoutte PM. Endothelium-dependent contractions in SHR: a tale of prostanoid TP and IP receptors. Br J Pharmacol 2009; 156: 563-74.

Bagi Z, Feher A, Casutto J, Akula K, Labinskyy N, Kaley G et al. Increased availability of angiotensin AT1 receptors leads to sustained arterial constriction to angiotensin II in diabetes role for Rho-kinase activation. Br J Pharmacol 2010; 163: 1059-68.

Tesfamariam B, Brown Ml, Deykın D, Cohen RA. Elevated Glucose Promotes Generation of Endothelium-derived Vasoconstrictor Prostanoids in Rabbit Aorta. J Clin Invest 1990; 85: 929-32.

De Mey JG, Vanhoutte PM. Heterogeneous behavior of the canine arterial and venous wall. Importance of the endothelium. Circ Res 1982; 51: 439-47.

De Mey JG, Vanhoutte PM. Control of vascular smooth muscle function by the endothelial cells. Gen Pharmacol 1983; 14: 39-41.

Miller VM, Vanhoutte PM. Endothelium-dependent contractions to arachidonic acid is mediated by products of cyclooxygenase. Am J Physiol 1985; 248: H432-7.

Katusic ZS, Shepherd JT, Vanhoutte PM. Endothelium-dependent contraction to stretch in canine basilar arteries. Am J Physiol 1987; 252: H671-3.

Yang D, Gluais P, Zhang JN, Vanhoutte PM, Feletou M. Endothelium dependent contractions to acetylcholine, ATP and the calcium ionophore A23187 in aortas from spontaneously hypertensive and normotensive rats. Fundam Clin Pharmacol 2004; 18: 321-6.

Iwama Y, Kato T, Muramatsu M, Asano H, Shimizu K, Toki Y, Miyazaki Y, Okumura K, Hashimoto H, Ito T. Correlation with blood pressure of the acetylcholine-induced endothelium-derived contracting factor in the rat aorta. Hypertension 1992; 19: 326-32.

Watt PA, Thurston H. Endothelium-dependent relaxation in resistance vessels from the spontaneously hypertensive rats. J Hypertens. 1989; 7: 661-6.

Líšková S, Petrová M, Karen P, Kuneš J, Zicha J. Effects of aging and hypertension on the participation of endothelium-derived constricting factor (EDCF) in norepinephrine-induced contraction of rat femoral artery. Eur J Pharmacol 2011; 667: 265-70.

Baretella O, Xu A, Vanhoutte PM. Acidosis prevents and alkalosis augments endothelium-dependent contractions in mousearteries. Pflugers Arch. 2014; 466: 295-305.

Rodriguez-Manas L, Angulo J, Vallejo S, Peiro C, Sanchez-Ferrer A,Cercas E, et al. Early and intermediate Amadori glycosylation adducts, oxidative stress, and endothelial dysfunction in the streptozotocin-induced diabetic rats vasculature. Diabetologia 2003; 46: 556-66.

Qian LB, Wang HP, Chen Y, Chen FX, Ma YY, Bruce LC, Xia Q. Luteolin reduces high glucose-mediated impairment of endothelium-dependent relaxation in rat aorta by reducing oxidative stress. Pharmacol Res 2010; 61: 281-7.

Shi Y, Vanhoutte PM. Oxidative stress and COX cause hyper-responsiveness in vascular smooth muscle of the femoral artery from diabetic rats. Br J Pharmacol 2008; 154: 639-51.

Vanhoutte PM, Shimokawa H , Feletou M, Tang EH. Endothelial dysfunction and vascular disease - a 30th anniversary update. Acta Physiol (Oxf.) 2017; 219: 22-96.

Affonso FDS, Cailleaux S, Pinto LFC, Gomes MDB, Tibirica E. Effects of high glucose concentrations on the endothelial function of the renal microcirculation of rabbits. Arq Bras Cardiol 2003; 81: 161-5.

Gross ER, Ladisa JF, Jr., Weihrauch D, Olson LE, Kress TT, Hettrick DA, Pagel PS, Warltier DC, Kersten JR. Reactive oxygen species modulate coronary wall shear stress and endothelial function during hyperglycemia. Am J Physiol Heart Circ Physiol 2003; 284: H1552-9.

Lash JM, Nase GP, Bohlen HG. Acute hyperglycemia depresses arteriolar NO formation in skeletal muscle. Am J Physiol 1999; 277: H1513-20.

Original Research