Naunyn-Schmiedeberg's Archives of Pharmacology

, Volume 391, Issue 11, pp 1221–1235 | Cite as

Activation of NQO-1 mediates the augmented contractions of isolated arteries due to biased activity of soluble guanylyl cyclase in their smooth muscle

  • Charlotte M. S. Detremmerie
  • Susan W. S. LeungEmail author
  • Paul M. Vanhoutte


Earlier studies on isolated arteries demonstrated that the para-quinone thymoquinone, like acute hypoxia, induces augmentation of contractions, depending on biased activity of soluble guanylyl cyclase (sGC), generating inosine-3′,5′-cyclic monophosphate (cyclic IMP) rather than guanosine-3′,5′-cyclic monophosphate (cyclic GMP). NAD(P)H:quinone oxidoreductase 1 (NQO-1), the enzyme responsible for biotransformation of quinones into hydroquinones, was examined for its involvement in these endothelium-dependent augmentations, establishing a link between the metabolism of quinones by NQO-1 and biased sGC activity. Isolated arteries of Sprague-Dawley rats (aortae and mesenteric arteries) and farm pigs (coronary arteries) were studied for measurement of changes in tension and collected to measure NQO-1 activity or its protein level. β-lapachone, an ortho-quinone and hence substrate of NQO-1, increased the activity of the enzyme and augmented contractions in arteries with endothelium. This augmentation was inhibited by endothelium removal and inhibitors of endothelial NO synthase (eNOS), sGC, or NQO-1; in preparations without endothelium or treated with an eNOS inhibitor, it was restored by the NO donor DETA NONOate and by ITP and cyclic IMP, revealing biased sGC activity as the underlying mechanism, as with thymoquinone. Hydroquinone, the end product of quinone metabolism by NQO-1, augmented contractions depending on sGC activation but in an endothelium-independent manner. In coronary arteries, repeated acute hypoxia caused similar augmentations as those to quinones that were inhibited by the NQO-1 inhibitor dicoumarol. Augmentations of contraction observed with different naturally occurring quinones and with acute hypoxia are initiated by quinone metabolism by NQO-1, in turn interfering with the NO/biased sGC pathway, suggesting a possibly detrimental role of this enzyme in ischemic cardiovascular disorders.


Vasoconstriction Biased soluble guanylyl cyclase (sGC) activity Cyclic IMP Quinones Hydroquinone Hypoxic augmentation NAD(P)H:quinone oxidoreductase 1 (NQO-1) 


Cyclic GMP

Guanosine-3′,5′-cyclic monophosphate

Cyclic IMP

Inosine-3′,5′-cyclic monophosphate


Diethylenetriamine NONOate






Endothelial nitric oxide synthase




Inosine triphosphate


Nω-nitro-l-arginine methyl ester hydrochloride


NAD(P)H:quinone oxidoreductase 1


Nitric oxide




Reactive oxygen species


Sodium dodecyl sulfate


Soluble guanylyl cyclase


Tris-buffered saline


1,3-Benzoxazol-2-yl-3-benzyl-3H-[1,2,3]triazolo[4,5-d]pyrimidin-7-yl sulfide



The authors would like to thank Ms. Yee Har Chung and Mr. Godfrey Man for their excellent technical assistance and Ms. Ivy Wong for her editorial and administrative support.

Author contribution

CD, SL, and PV conceived and designed the research. CD conducted the experiments and analyzed the data. CD, SL, and PV wrote the manuscript. All authors read and approved the final version of the manuscript.


This work was supported by the Health and Medical Research Fund [15162231] of the Food and Health Bureau of the Government of the Hong Kong Special Administrative Region and by the General Research Fund [17112914] of the Hong Kong Research Grant Council.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted.

Supplementary material

210_2018_1548_MOESM1_ESM.pdf (373 kb)
ESM 1 (PDF 372 kb)


  1. Abukhader MM (2013) Thymoquinone in the clinical treatment of cancer: fact or fiction? Pharmacogn Rev 7:117–120CrossRefGoogle Scholar
  2. Al-Awqati Q (1999) One hundred years of membrane permeability: does Overton still rule? Nat Cell Biol 1(8):E201–E202CrossRefGoogle Scholar
  3. Amin B, Hosseinzadeh H (2016) Black cumin (Nigella sativa) and its active constituent thymoquinone: an overview on the analgesic and anti-inflammatory effects. Planta Med 82(1–2):8–16PubMedGoogle Scholar
  4. Beste KY, Burhenne H, Kaever V, Stasch JP, Seifert R (2012) Nucleotidyl cyclase activity of soluble guanylyl cyclase alpha1beta1. Biochemistry 51:194–204CrossRefGoogle Scholar
  5. Bey EA, Wuerzberger-Davis SM, Pink JJ, Yang CR, Araki S, Reinicke KE, Bentle MS, Dong Y, Cataldo E, Criswell TL, Wagner MW, Li L, Gao J, Boothman DA (2006) Mornings with art, lessons learned: feedback regulation, restriction threshold biology, and redundancy govern molecular stress responses. J Cell Physiol 209:604–610CrossRefGoogle Scholar
  6. Bing RJ, Saeed M, Hartmann A (1987) The vasodilator effect of coronary vascular endothelium in situ: its inactivation by hydroquinone. J Mol Cell Cardiol 19:343–348CrossRefGoogle Scholar
  7. Black SR, Fennell TR, Mathews JM, Snyder RW, Patel PR, Watson SL et al (2017) Disposition of [14C]hydroquinone in Harlan Sprague-Dawley rats and B6C3F1/N mice: species and route comparison. Xenobiot 22:1–14Google Scholar
  8. Butt M, Khair OA, Dwivedi G, Shantsila A, Shantsila E, Lip GY (2011) Myocardial perfusion by myocardial contrast echocardiography and endothelial dysfunction in obstructive sleep apnea. Hypertension 58:417–424CrossRefGoogle Scholar
  9. Capra V, Back M, Angiolillo DJ, Cattaneo M, Sakariassen KS (2014) Impact of vascular thromboxane prostanoid receptor activation on hemostasis, thrombosis, oxidative stress, and inflammation. J Thromb Haemost 12:126–137CrossRefGoogle Scholar
  10. Chan CK, Mak J, Gao Y, Man RY, Vanhoutte PM (2011) Endothelium-derived NO, but not cyclic GMP, is required for hypoxic augmentation in isolated porcine coronary arteries. Am J Physiol Heart Circ Physiol 301:H2313–H2321CrossRefGoogle Scholar
  11. Chen Z, Zhang X, Ying L, Dou D, Li Y, Bai Y, Liu J, Liu L, Feng H, Yu X, Leung SWS, Vanhoutte PM, Gao Y (2014) cIMP synthesized by sGC as a mediator of hypoxic contraction of coronary arteries. Am J Physiol Heart Circ Physiol 307:H328–H336CrossRefGoogle Scholar
  12. Choi EK, Terai K, Ji IM, Kook YH, Park KH, Oh ET, Griffin RJ, Lim BU, Kim JS, Lee DS, Boothman DA, Loren M, Song CW, Park HJ (2007) Upregulation of NAD(P)H:quinone oxidoreductase by radiation potentiates the effect of bioreductive beta-lapachone on cancer cells. Neoplasia 9:634–642CrossRefGoogle Scholar
  13. Detremmerie CM, Chen Z, Li Z, Alkharfy KM, Leung SW, Xu A et al (2016) Endothelium-dependent contractions of isolated arteries to thymoquinone require biased activity of sGC with subsequent cIMP production. J Pharmacol Exp Ther 358:558–568CrossRefGoogle Scholar
  14. Ernster L, Dallner G (1995) Biochemical, physiological and medical aspects of ubiquinone function. Biochim Biophys Acta 1271(1):195–204CrossRefGoogle Scholar
  15. Gao Y, Chen Z, Leung SW, Vanhoutte PM (2015) Hypoxic vasospasm mediated by cIMP: when soluble guanylyl cyclase turns bad. J Cardiovasc Pharmacol 65:545–548CrossRefGoogle Scholar
  16. Gao Y (2016) Conventional and unconventional mechanisms for soluble guanylyl cyclase signaling. J Cardiovasc Pharmacol 67(5):367–372CrossRefGoogle Scholar
  17. Garthwaite J, Southam E, Boulton CL, Nielsen EB, Schmidt K, Mayer B (1995) Potent and selective inhibition of nitric oxide-sensitive guanylyl cyclase by 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one. Mol Pharmacol 48:184–188PubMedGoogle Scholar
  18. Gassmann M, Grenacher B, Rohde B, Vogel J (2009) Quantifying Western blots: pitfalls of densitometry. Electrophoresis 30:1845–1855CrossRefGoogle Scholar
  19. Ghayur MN, Gilani AH, Janssen LJ (2012) Intestinal, airway, and cardiovascular relaxant activities of thymoquinone. Evid Based Complement Alternat Med 2012:305319CrossRefGoogle Scholar
  20. Gräser T, Vanhoutte PM (1991) Hypoxic contraction of canine coronary arteries: role of endothelium and cGMP. Am J Physiol Heart Circ Physiol 261(6 Pt 2):H1769–H1777CrossRefGoogle Scholar
  21. Gupta SC, Patchva S, Aggarwal BB (2013) Therapeutic roles of curcumin: lessons learned from clinical trials. AAPS J 15:195–218CrossRefGoogle Scholar
  22. Gustafson DL, Siegel D, Rastatter JC, Merz AL, Parpal JC, Kepa JK, Ross D, Long ME (2003) Kinetics of NAD(P)H:quinone oxidoreductase I (NQO1) inhibition by Mitomycin C in vitro and in vivo. J Pharmacol Exp Ther 305:1079–1086CrossRefGoogle Scholar
  23. Idris-Khodja N, Schini-Kerth V (2012) Thymoquinone improves aging-related endothelial dysfunction in the rat mesenteric artery. Naunyn Schmiedeberg's Arch Pharmacol 385:749–758CrossRefGoogle Scholar
  24. Igarashi Y, Tamura Y, Suzuki K, Tanabe Y, Yamaguchi T, Fujita T, Yamazoe M, Aizawa Y, Shibata A (1993) Coronary artery spasm is a major cause of sudden cardiac arrest in survivors without underlying heart disease. Coron Artery Dis 4(2):177–185CrossRefGoogle Scholar
  25. Jolly SR, Kane WJ, Bailie MB, Abrams GD, Lucchesi BR (1984). Canine myocardial reperfusion injury. Its reduction by the combined administration of superoxide dismutase and catalase. Circ Res 54(3):277-285CrossRefGoogle Scholar
  26. Khattab MM, Nagi MN (2007) Thymoquinone supplementation attenuates hypertension and renal damage in nitric oxide deficient hypertensive rats. Phytother Res 21(5):410–414CrossRefGoogle Scholar
  27. Kim YH, Hwang JH, Noh JR, Gang GT, Kim DH, Son HY, Kwak TH, Shong M, Lee IK, Lee CH (2011) Activation of NAD(P)H:quinone oxidoreductase ameliorates spontaneous hypertension in an animal model via modulation of eNOS activity. Cardiovasc Res 91:519–527CrossRefGoogle Scholar
  28. Kim YH, Hwang JH, Kim KS, Noh JR, Gang GT, Seo Y, Nam KH, Kwak TH, Lee HG, Lee CH (2015a) NAD(P)H:quinone oxidoreductase 1 activation reduces blood pressure through regulation of endothelial nitric oxide synthase acetylation in spontaneously hypertensive rats. Am J Hypertens 28:50–57CrossRefGoogle Scholar
  29. Kim I, Kim H, Ro J, Jo K, Karki S, Khadka P, Yun G, Lee J (2015b) Preclinical pharmacokinetic evaluation of β-lapachone: characteristics of oral bioavailability and first-pass metabolism in rats. Biomol Ther (Seoul) 23(3):296–300CrossRefGoogle Scholar
  30. Lee CH, Khoo SM, Chan MY, Wong HB, Low AF, Phua QH, Richards AM, Tan HC, Yeo TC (2011) Severe obstructive sleep apnea and outcomes following myocardial infarction. J Clin Sleep Med 7:616–621PubMedPubMedCentralGoogle Scholar
  31. Lee MY, Tse HF, Siu CW, Zhu SG, Man RY, Vanhoutte PM (2007) Genomic changes in regenerated porcine coronary arterial endothelial cells. Arterioscler Throm Vasc Biol 27(11):2443–2449CrossRefGoogle Scholar
  32. Leung GP, Man RY, Tse CM (2005) d-Glucose upregulates adenosine transport in cultured human aortic smooth muscle cells. Am J Physiol Heart Circ Physiol 288:H2756–H2762CrossRefGoogle Scholar
  33. Li RW, Yang C, Sit AS, Lin SY, Ho EY, Leung GP (2012) Physiological and pharmacological roles of vascular nucleoside transporters. J Cardiovasc Pharmacol 59:10–15CrossRefGoogle Scholar
  34. Mendelsohn ME, Karas RH (2005) Molecular and cellular basis of cardiovascular gender differences. Science 308(5728):1583–1587CrossRefGoogle Scholar
  35. Messerli FH, Garavaglia GE, Schmieder RE, Sundgaard-Rijse K, Nunez BD, Amodeo C (1987) Disparate cardiovascular findings in men and women with essential hypertension. Ann Intern Med 107(2):158–161CrossRefGoogle Scholar
  36. Nasongkla N, Wiedmann AF, Bruening A, Beman M, Ray D, Bornmann WG, Boothman DA, Gao J (2003) Enhancement of solubility and bioavailability of beta-lapachone using cyclodextrin inclusion complexes. Pharm Res 20:1626–1633CrossRefGoogle Scholar
  37. Oh ET, Park HJ (2015) Implications of NQO1 in cancer therapy. BMB Rep 48:609–617CrossRefGoogle Scholar
  38. Papaharalambus CA, Griendling KK (2007) Basic mechanisms of oxidative stress and reactive oxygen species in cardiovascular injury. Trends Cardiovasc Med 17:48–54CrossRefGoogle Scholar
  39. Pathan SA, Jain GK, Zaidi SM, Akhter S, Vohora D, Chander P et al (2011) Stability-indicating ultra-performance liquid chromatography method for the estimation of thymoquinone and its application in biopharmaceutical studies. Biomed Chromatogr 25(5):613–620CrossRefGoogle Scholar
  40. Pearson PJ, Lin PJ, Schaff HV, Vanhoutte PM (1996) Augmented endothelium-dependent constriction to hypoxia early and late following reperfusion of the canine coronary artery. Clin Exp Pharmacol Physiol 23:634–641CrossRefGoogle Scholar
  41. Pink JJ, Planchon SM, Tagliarino C, Varne ME, Siegel D, Boothman DA (2000) NAD(P)H:Quinone oxidioreductase activity is the principal determinant of beta-lapachone cytotoxicity. J Biol Chem 275:5416–5424CrossRefGoogle Scholar
  42. Rees DD, Palmer RM, Schulz R, Hodson HF, Moncada S (1990) Characterization of three inhibitors of endothelial nitric oxide synthase in vitro and in vivo. Br J Pharmacol 101:746–752CrossRefGoogle Scholar
  43. Ross D, Kepa JK, Winski SL, Beall HD, Anwar A, Siegel D (2000) NAD(P)H:quinone oxidoreductase 1 (NQO1): chemoprotection, bioactivation, gene regulation and genetic polymorphisms. Chem Biol Interact 129:77–97CrossRefGoogle Scholar
  44. Salmani JM, Asghar S, Lv H, Zhou J (2014) Aqueous solubility and degradation kinetics of the phytochemical anticancer thymoquinone; probing the effects of solvents, pH and light. Molecule 19:5925–5939CrossRefGoogle Scholar
  45. Santos-Parker J, Strahler TR, Bassett CJ, Bispham NZ, Chonchol MB, Seals DR (2017) Curcumin supplementation improves vascular endothelial function in healthy middle-aged and older adults by increasing nitric oxide bioavailability and reducing oxidative stress. Aging 9:187–205CrossRefGoogle Scholar
  46. Scott KA, Barnes J, Whitehead RC, Stratford IJ, Nolan KA (2011) Inhibitors of NQO1: identification of compounds more potent than dicoumarol without associated off-target effects. Biochem Pharmacol 81(3):355–363CrossRefGoogle Scholar
  47. Shi Y, So KF, Man RY, Vanhoutte PM (2007) Oxygen-derived free radicals mediate endothelium-dependent contractions in femoral arteries of rats with streptozotocin-induced diabetes. Br J Pharmacol 152:1033–1041CrossRefGoogle Scholar
  48. Shi Y, Vanhoutte PM (2008) Oxidative stress and COX cause hyper-responsiveness in vascular smooth muscle of the femoral artery from diabetic rats. Br J Pharmacol 154:639–651CrossRefGoogle Scholar
  49. Siegel D, Yan C, Ross D (2012) NAD(P)H:quinone oxidoreductase 1 (NQO1) in the sensitivity and resistance to antitumor quinones. Biochem Pharmacol 83:1033–1040CrossRefGoogle Scholar
  50. Slavich M, Patel RS (2016) Coronary artery spasm: current knowledge and residual uncertainties. Int J Cardiol Heart Vasc 10:47–53PubMedPubMedCentralGoogle Scholar
  51. Suddek GM (2010) Thymoquinone-induced relaxation of isolated rat pulmonary artery. J Ethnopharmacol 127:210–214CrossRefGoogle Scholar
  52. Tang EH, Leung FP, Huang Y, Félétou M, So KF, Man RY et al (2007) Calcium and reactive oxygen species increase in endothelial cells in response to releasers of endothelium-derived contracting factor. Br J Pharmacol 151:15–23CrossRefGoogle Scholar
  53. Timson DJ (2017) Dicoumarol: a drug which hits at least two very different targets in vitamin K metabolism. Curr Drug Targets 18:500–510CrossRefGoogle Scholar
  54. Tsvetkov P, Asher G, Reiss V, Shaul Y, Sachs L, Lotem J (2005) Inhibition of NAD(P)H:quinone oxidoreductase 1 activity and induction of p53 degradation by the natural phenolic compound curcumin. Proc Natl Acad Sci U S A 102:5535–5540CrossRefGoogle Scholar
  55. Vanhoutte PM, Leusen I (1969) The reactivity of isolated venous preparations to electrical stimulation. Pflugers Arch 306:341–353CrossRefGoogle Scholar
  56. Vanhoutte PM, Tang EH (2008) Endothelium-dependent contractions: when a good guy turns bad! J Physiol 586:5295–5304CrossRefGoogle Scholar
  57. Wolter S, Golombek M, Seifert R (2011) Differential activation of cAMP- and cGMP-dependent protein kinases by cyclic purine and pyrimidine nucleotides. Biochem Biophys Res Commun 415:563–566CrossRefGoogle Scholar
  58. 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 104:228–235CrossRefGoogle Scholar
  59. Zhu H, Jia Z, Mahaney JE, Ross D, Misra HP, Trush MA, Li Y (2007) The highly expressed and inducible endogenous NAD(P)H:quinone oxidoreductase 1 in cardiovascular cells acts as a potential superoxide scavenger. Cardiovasc Toxicol 7:202–211CrossRefGoogle Scholar
  60. Zhu H, Li Y (2012) NAD(P)H:quinone oxidoreductase 1 and its potential protective role in cardiovascular diseases and related conditions. Cardiovasc Toxicol 12:39–45CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Pharmacology and Pharmacy and State Key Laboratory for Pharmaceutical Biotechnology, Li Ka Shing Faculty of MedicineThe University of Hong KongHong KongChina

Personalised recommendations