, 37:79 | Cite as

Prostanoid-mediated contractions of the carotid artery become Nox2-independent with aging

  • Matthias R. Meyer
  • Natalie C. Fredette
  • Matthias Barton
  • Eric R. Prossnitz


Aging is a major risk factor for carotid artery disease that may lead to stroke and dementia. Vascular effects associated with aging include increased vasomotor tone, as well as enhanced contractility to endothelial vasoconstrictor prostanoids and reduced nitric oxide (NO) bioactivity partly due to increased oxidative stress. We hypothesized that vascular NADPH oxidase (Nox)-derived superoxide may be involved in prostanoid- and NO-related functional aging. NO-mediated relaxations and prostanoid-mediated contractions to acetylcholine as well as phenylephrine-dependent contractions were investigated in the carotid artery from young (4 months) and aged mice (24 months). Gene expression of Nox subunits and endothelial NO synthase (eNOS) was determined in the carotid artery and aorta. In young mice, the thromboxane-prostanoid receptor antagonist SQ 29,548 fully blocked acetylcholine-induced contractions while reducing responses to phenylephrine by 75 %. The Nox2-targeted inhibitor Nox2ds-tat and the superoxide scavenger tempol reduced acetylcholine-stimulated, prostanoid-mediated contractions by 85 and 75 %, respectively, and phenylephrine-dependent contractions by 45 %. Unexpectedly, in aged mice, the substantial Nox2-dependent component of acetylcholine- and phenylephrine-induced, prostanoid-mediated contractions was abolished. In addition, endothelium-dependent, NO-mediated relaxations were impaired with aging. The expression of Nox subunits was greater in the aorta compared with the carotid artery, in which Nox1 was undetectable. eNOS gene expression was reduced in the aorta of aged compared to young mice. In conclusion, aging decreases prostanoid-mediated contractility in the carotid artery involving a loss of Nox2 activity and is associated with impaired endothelium-dependent, NO-mediated relaxation. These findings may contribute to a better understanding of the pathophysiology of carotid artery disease and the aging process.


Aging Carotid artery NADPH oxidase Nox2 Prostanoid Superoxide 



We thank Dr. Chelin Hu and Daniel F. Cimino for expert technical assistance. This study was supported by the National Institutes of Health (R01 CA127731 and CA163890 to E.R.P.), Dedicated Health Research Funds from the University of New Mexico School of Medicine allocated to the Signature Program in Cardiovascular and Metabolic Diseases (to E.R.P.), and the Swiss National Science Foundation (grants 135874 and 141501 to M.R.M. and grants 108258 and 122504 to M.B.). N.C.F. was supported by NIH training grant HL07736.


  1. Barton M (2014) Aging and endothelin: determinants of disease. Life Sci 118:97–109CrossRefPubMedGoogle Scholar
  2. Barton M, Cosentino F, Brandes RP, Moreau P, Shaw S, Luscher TF (1997) Anatomic heterogeneity of vascular aging: role of nitric oxide and endothelin. Hypertension 30:817–824CrossRefPubMedGoogle Scholar
  3. Brandes RP, Weissmann N, Schroder K (2010) NADPH oxidases in cardiovascular disease. Free Radic Biol Med 49:687–706CrossRefPubMedGoogle Scholar
  4. Cosentino F, Sill JC, Katusic ZS (1994) Role of superoxide anions in the mediation of endothelium-dependent contractions. Hypertension 23:229–235CrossRefPubMedGoogle Scholar
  5. Crauwels HM, Van Hove CE, Herman AG, Bult H (2000) Heterogeneity in relaxation mechanisms in the carotid and the femoral artery of the mouse. Eur J Pharmacol 404:341–351CrossRefPubMedGoogle Scholar
  6. Csanyi G, Cifuentes-Pagano E, Al Ghouleh I, Ranayhossaini DJ, Egana L, Lopes LR, Jackson HM, Kelley EE, Pagano PJ (2011) Nox2 B-loop peptide, Nox2ds, specifically inhibits the NADPH oxidase Nox2. Free Radic Biol Med 51:1116–1125PubMedCentralCrossRefPubMedGoogle Scholar
  7. Dalager S, Paaske WP, Kristensen IB, Laurberg JM, Falk E (2007) Artery-related differences in atherosclerosis expression: implications for atherogenesis and dynamics in intima-media thickness. Stroke 38:2698–2705CrossRefPubMedGoogle Scholar
  8. DeLean A, Munson PJ, Rodbard D (1978) Simultaneous analysis of families of sigmoidal curves: application to bioassay, radioligand assay, and physiological dose-response curves. Am J Physiol 235:E97–E102PubMedGoogle Scholar
  9. Donato AJ, Eskurza I, Silver AE, Levy AS, Pierce GL, Gates PE, Seals DR (2007) Direct evidence of endothelial oxidative stress with aging in humans: relation to impaired endothelium-dependent dilation and upregulation of nuclear factor-kappaB. Circ Res 100:1659–1666CrossRefPubMedGoogle Scholar
  10. Drummond GR, Selemidis S, Griendling KK, Sobey CG (2011) Combating oxidative stress in vascular disease: NADPH oxidases as therapeutic targets. Nat Rev Drug Discov 10:453–471PubMedCentralCrossRefPubMedGoogle Scholar
  11. Durrant JR, Seals DR, Connell ML, Russell MJ, Lawson BR, Folian BJ, Donato AJ, Lesniewski LA (2009) Voluntary wheel running restores endothelial function in conduit arteries of old mice: direct evidence for reduced oxidative stress, increased superoxide dismutase activity and down-regulation of NADPH oxidase. J Physiol 587:3271–3285PubMedCentralCrossRefPubMedGoogle Scholar
  12. Feletou M, Vanhoutte PM (2006) Endothelial dysfunction: a multifaceted disorder (The Wiggers Award Lecture). Am J Physiol Heart Circ Physiol 291:H985–H1002CrossRefPubMedGoogle Scholar
  13. Fleenor BS, Seals DR, Zigler ML, Sindler AL (2012) Superoxide-lowering therapy with TEMPOL reverses arterial dysfunction with aging in mice. Aging Cell 11:269–276PubMedCentralCrossRefPubMedGoogle Scholar
  14. Gao YJ, Lee RM (2005) Hydrogen peroxide is an endothelium-dependent contracting factor in rat renal artery. Br J Pharmacol 146:1061–1068PubMedCentralCrossRefPubMedGoogle Scholar
  15. Gerland P, Raftery AE, Sevcikova H, Li N, Gu D, Spoorenberg T, Alkema L, Fosdick BK, Chunn J, Lalic N, Bay G, Buettner T, Heilig GK, Wilmoth J (2014) World population stabilization unlikely this century. Science 346:234–237PubMedCentralCrossRefPubMedGoogle Scholar
  16. Gorelick PB, Scuteri A, Black SE, Decarli C, Greenberg SM, Iadecola C, Launer LJ, Laurent S, Lopez OL, Nyenhuis D, Petersen RC, Schneider JA, Tzourio C, Arnett DK, Bennett DA, Chui HC, Higashida RT, Lindquist R, Nilsson PM, Roman GC, Sellke FW, Seshadri S (2011) Vascular contributions to cognitive impairment and dementia: a statement for healthcare professionals from the american heart association/american stroke association. Stroke 42:2672–2713PubMedCentralCrossRefPubMedGoogle Scholar
  17. Hahn NE, Musters RJ, Fritz JM, Pagano PJ, Vonk AB, Paulus WJ, van Rossum AC, Meischl C, Niessen HW, Krijnen PA (2014) Early NADPH oxidase-2 activation is crucial in phenylephrine-induced hypertrophy of H9c2 cells. Cell Signal 26:1818–1824PubMedCentralCrossRefPubMedGoogle Scholar
  18. Heymes C, Habib A, Yang D, Mathieu E, Marotte F, Samuel J, Boulanger CM (2000) Cyclo-oxygenase-1 and -2 contribution to endothelial dysfunction in ageing. Br J Pharmacol 131:804–810PubMedCentralCrossRefPubMedGoogle Scholar
  19. Huisman A, Van De Wiel A, Rabelink TJ, Van Faassen EE (2004) Wine polyphenols and ethanol do not significantly scavenge superoxide nor affect endothelial nitric oxide production. J Nutr Biochem 15:426–432CrossRefPubMedGoogle Scholar
  20. Judkins CP, Diep H, Broughton BR, Mast AE, Hooker EU, Miller AA, Selemidis S, Dusting GJ, Sobey CG, Drummond GR (2010) Direct evidence of a role for Nox2 in superoxide production, reduced nitric oxide bioavailability, and early atherosclerotic plaque formation in ApoE-/- mice. Am J Physiol Heart Circ Physiol 298:H24–H32CrossRefPubMedGoogle Scholar
  21. Katusic ZS, Schugel J, Cosentino F, Vanhoutte PM (1993) Endothelium-dependent contractions to oxygen-derived free radicals in the canine basilar artery. Am J Physiol 264:H859–H864PubMedGoogle Scholar
  22. Kretz M, Mundy AL, Widmer CC, Barton M (2006) Early aging and anatomic heterogeneity determine cyclooxygenase-mediated vasoconstriction to angiotensin II in mice. J Cardiovasc Pharmacol 48:30–33CrossRefPubMedGoogle Scholar
  23. Lassegue B, San Martin A, Griendling KK (2012) Biochemistry, physiology, and pathophysiology of NADPH oxidases in the cardiovascular system. Circ Res 110:1364–1390PubMedCentralCrossRefPubMedGoogle Scholar
  24. Ling X, Cota-Gomez A, Flores NC, Hernandez-Saavedra D, McCord JM, Marecki JC, Haskins K, McDuffie M, Powers K, Kench J, Oka M, McMurtry I, Flores SC (2005) Alterations in redox homeostasis and prostaglandins impair endothelial-dependent vasodilation in euglycemic autoimmune nonobese diabetic mice. Free Radic Biol Med 39:1089–1098CrossRefPubMedGoogle Scholar
  25. Mehta PK, Griendling KK (2007) Angiotensin II cell signaling: physiological and pathological effects in the cardiovascular system. Am J Physiol Cell Physiol 292:C82–C97CrossRefPubMedGoogle Scholar
  26. Meyer MR, Fredette NC, Barton M, Prossnitz ER (2013) Regulation of vascular smooth muscle tone by adipose-derived contracting factor. PLoS One 8, e79245PubMedCentralCrossRefPubMedGoogle Scholar
  27. Meyer MR, Barton M, Prossnitz ER (2014a) Functional heterogeneity of NADPH oxidase-mediated contractions to endothelin with vascular aging. Life Sci 118:226–231CrossRefPubMedGoogle Scholar
  28. Meyer MR, Fredette N, Barton M, Prossnitz ER (2014b) Endothelin-1 but not angiotensin II contributes to functional aging in murine carotid arteries. Life Sci 118:213–218CrossRefPubMedGoogle Scholar
  29. Modrick ML, Kinzenbaw DA, Chu Y, Sigmund CD, Faraci FM (2012) Peroxisome proliferator-activated receptor-gamma protects against vascular aging. Am J Physiol Regul Integr Comp Physiol 302:R1184–R1190PubMedCentralCrossRefPubMedGoogle Scholar
  30. Mukai Y, Shimokawa H, Higashi M, Morikawa K, Matoba T, Hiroki J, Kunihiro I, Talukder HM, Takeshita A (2002) Inhibition of renin-angiotensin system ameliorates endothelial dysfunction associated with aging in rats. Arterioscler Thromb Vasc Biol 22:1445–1450CrossRefPubMedGoogle Scholar
  31. Novella S, Dantas AP, Segarra G, Novensa L, Heras M, Hermenegildo C, Medina P (2013) Aging enhances contraction to thromboxane A2 in aorta from female senescence-accelerated mice. Age (Dordr) 35:117–128CrossRefGoogle Scholar
  32. Oudot A, Martin C, Busseuil D, Vergely C, Demaison L, Rochette L (2006) NADPH oxidases are in part responsible for increased cardiovascular superoxide production during aging. Free Radic Biol Med 40:2214–2222CrossRefPubMedGoogle Scholar
  33. Pereira AC, Olivon VC, de Oliveira AM (2010) An apparent paradox: attenuation of phenylephrine-mediated calcium mobilization and hyperreactivity to phenylephrine in contralateral carotid after balloon injury. J Cardiovasc Pharmacol 56:162–170CrossRefPubMedGoogle Scholar
  34. Rey FE, Cifuentes ME, Kiarash A, Quinn MT, Pagano PJ (2001) Novel competitive inhibitor of NAD(P)H oxidase assembly attenuates vascular O(2)(-) and systolic blood pressure in mice. Circ Res 89:408–414CrossRefPubMedGoogle Scholar
  35. Seals DR, Jablonski KL, Donato AJ (2011) Aging and vascular endothelial function in humans. Clin Sci (Lond) 120:357–375CrossRefGoogle Scholar
  36. Shi Y, Man RY, Vanhoutte PM (2008) Two isoforms of cyclooxygenase contribute to augmented endothelium-dependent contractions in femoral arteries of 1-year-old rats. Acta Pharmacol Sin 29:185–192CrossRefPubMedGoogle Scholar
  37. Solberg LA, Eggen DA (1971) Localization and sequence of development of atherosclerotic lesions in the carotid and vertebral arteries. Circulation 43:711–724CrossRefPubMedGoogle Scholar
  38. Tang EH, Leung FP, Huang Y, Feletou M, So KF, Man RY, Vanhoutte PM (2007) Calcium and reactive oxygen species increase in endothelial cells in response to releasers of endothelium-derived contracting factor. Br J Pharmacol 151:15–23PubMedCentralCrossRefPubMedGoogle Scholar
  39. Traupe T, Lang M, Goettsch W, Munter K, Morawietz H, Vetter W, Barton M (2002) Obesity increases prostanoid-mediated vasoconstriction and vascular thromboxane receptor gene expression. J Hypertens 20:2239–2245CrossRefPubMedGoogle Scholar
  40. Vanhoutte PM (2013) One or two, does it matter as long as the arterial wall is coxygenated? Hypertension 62:244–246CrossRefPubMedGoogle Scholar
  41. Wang G, Sarkar P, Peterson JR, Anrather J, Pierce JP, Moore JM, Feng J, Zhou P, Milner TA, Pickel VM, Iadecola C, Davisson RL (2013) COX-1-derived PGE2 and PGE2 type 1 receptors are vital for angiotensin II-induced formation of reactive oxygen species and Ca(2+) influx in the subfornical organ. Am J Physiol Heart Circ Physiol 305:H1451–H1461PubMedCentralCrossRefPubMedGoogle Scholar
  42. Wong SL, Leung FP, Lau CW, Au CL, Yung LM, Yao X, Chen ZY, Vanhoutte PM, Gollasch M, Huang Y (2009) Cyclooxygenase-2-derived prostaglandin F2alpha mediates endothelium-dependent contractions in the aortae of hamsters with increased impact during aging. Circ Res 104:228–235CrossRefPubMedGoogle Scholar
  43. Yang D, Feletou M, Boulanger CM, Wu HF, Levens N, Zhang JN, Vanhoutte PM (2002) Oxygen-derived free radicals mediate endothelium-dependent contractions to acetylcholine in aortas from spontaneously hypertensive rats. Br J Pharmacol 136:104–110PubMedCentralCrossRefPubMedGoogle Scholar
  44. Zhang M, Song P, Xu J, Zou MH (2011) Activation of NAD(P)H oxidases by thromboxane A2 receptor uncouples endothelial nitric oxide synthase. Arterioscler Thromb Vasc Biol 31:125–132PubMedCentralCrossRefPubMedGoogle Scholar
  45. Zhou Y, Varadharaj S, Zhao X, Parinandi N, Flavahan NA, Zweier JL (2005) Acetylcholine causes endothelium-dependent contraction of mouse arteries. Am J Physiol Heart Circ Physiol 289:H1027–H1032CrossRefPubMedGoogle Scholar
  46. Zieman SJ, Melenovsky V, Kass DA (2005) Mechanisms, pathophysiology, and therapy of arterial stiffness. Arterioscler Thromb Vasc Biol 25:932–943CrossRefPubMedGoogle Scholar

Copyright information

© American Aging Association 2015

Authors and Affiliations

  • Matthias R. Meyer
    • 1
    • 2
  • Natalie C. Fredette
    • 1
  • Matthias Barton
    • 3
  • Eric R. Prossnitz
    • 1
  1. 1.Department of Internal MedicineUniversity of New Mexico Health Sciences CenterAlbuquerqueUSA
  2. 2.Department of CardiologyCantonal HospitalAarauSwitzerland
  3. 3.Molecular Internal MedicineUniversity of ZürichZürichSwitzerland

Personalised recommendations