Cobalt-Induced Hypercontraction is Mediated by Generationof Reactive Oxygen Species and Influx of Calcium in Isolated RatAorta

  • Shahnawaz Ahmad Wani
  • Luqman Ahmad Khan
  • Seemi Farhat BasirEmail author


To investigate the mechanism of cobalt-mediated phenylephrine (PE)-induced contraction in endothelium-intact isolated Wistar rat aortic rings. Effect of dose-dependent concentrations of cobalt on PE-induced contraction was investigated in isolated Wistar rat aortic rings using an organ bath system. Aortic rings were pre-incubated with verapamil (1 μM and 20 μM), gadolinium, apocynin, indomethacin or N-G-nitro-l-arginine methyl ester (L-NAME) separately before incubation with cobalt. Endothelium-intact aortic rings were incubated with 800 nM, 1 μM, 10 μM, 50 μM cobalt; we observed 20%, 22%, 32% and 27% increased contractions respectively, while no effect was seen in tension recording on cobalt exposure. Incubation of endothelium-intact aortic rings with 100 μM apocynin and 100 μM L-NAME suggested the role of NADPH oxidase in generation of reactive oxygen species (ROS) and decrease in bioavailability of nitric oxide (NO) from eNOS on exposure to cobalt. Aortic rings pre-incubated with 1 μM and 20 μM verapamil suggested role of both L-type and T-type calcium channels in influx of extracellular calcium in smooth muscle cells. We observed no role of store-operated calcium channels (SOCC) in calcium influx due to cobalt exposure and cyclooxygenase in generation of prostanoids in isolated aortic rings. Cobalt caused rise of PE-induced contractions as a result of the endothelial generation of ROS, by decreasing bioavailability of NO. Generation of ROS may be responsible for causing the influx of extracellular calcium through L-type and T-type Ca2+ channels in smooth muscle cells.


Cobalt Vasocontraction Reactive oxygen species Calcium channels Nitric oxide 



  1. 1.
    O’ Leary F, Samman S (2012) Vitamin B12 in health and disease. Nutrients 2(3):299–316CrossRefGoogle Scholar
  2. 2.
    Nordberg GF, Fowler BA, Nordberg M (2014) Handbook on the toxicology of metals. Academic PressGoogle Scholar
  3. 3.
    Kumagai S, Kusaka Y, Goto S (1996) Cobalt exposure level and variability in the hard metal industry of Japan. Am Ind Hyg Assoc 57(4):365–369CrossRefGoogle Scholar
  4. 4.
    Polyzois I, Nikolopoulos D, Michos I, Patsouris E, Theocharis S (2012) Local and systemic toxicity of nanoscale debris particles in total hip arthroplasty. J Appl Toxicol 32(4):255–269CrossRefGoogle Scholar
  5. 5.
    Sampson B, Hart A (2012) Clinical usefulness of blood metal measurements to assess the failure of metal-on-metal hip implants. Ann Clin Biochem 49(2):118–131CrossRefGoogle Scholar
  6. 6.
    Ichikawa Y, Kusaka Y, Goto S (1985) Biological monitoring of cobalt exposure, based on cobalt concentrations in blood and urine. Int Arch Occup Environ Health 55(4):269–276CrossRefGoogle Scholar
  7. 7.
    Mohiuddin SM, Taskar PK, Rheault M, Roy P-E, Chenard J, Morin Y (1970) Experimental cobalt cardiomyopathy. Am Heart J 80(4):532–543CrossRefGoogle Scholar
  8. 8.
  9. 9.
    Edel J, Pozzi G, Sabbioni E, Pietra R, Devos S (1994) Metabolic and toxicological studies on cobalt. Sci Total Environ 150(1–3):233–244CrossRefGoogle Scholar
  10. 10.
    Shibata S, Kurahashi K, Kuchii M (1973) A possible etiology of contractility impairment of vascular smooth muscle from spontaneously hypertensive rats. J Pharmacol Exp Ther 185(2):406–417Google Scholar
  11. 11.
    Bohr DF (1974) Reactivity of vascular smooth muscle from normal and hypertensive rats: effect of several cations. Fed Proc 33:127Google Scholar
  12. 12.
    Gallagher MJ, Alade PI, Dominiczak AF, Bohr DF (1994) Cobalt contraction of vascular smooth muscle is calcium dependent. J Cardiovasc Pharmacol 24(2):293–297CrossRefGoogle Scholar
  13. 13.
    Seong Y, Kim E, Park T-G, Seok Y, Baek W, Kim S-O, Lim DG, Yang DH, Kim I (2005) Endothelial dysfunction after exposure to cobalt chloride enhanced vascular contractility. Environ Toxicol Pharmacol 20(2):297–304CrossRefGoogle Scholar
  14. 14.
    Kawahara Y, Tanonaka K, Daicho T, Nawa M, Oikawa R, Nasa Y, Takeo S (2005) Preferable anesthetic conditions for echocardiographic determination of murine cardiac function. J Pharmacol Sci 99(1):95–104CrossRefGoogle Scholar
  15. 15.
    Shabir H, Kundu S, Basir SF, Khan LA (2014) Modulation of Pb (II) caused aortal constriction by eugenol and carvacrol. Biol Trace Elem Res 161(1):116–122CrossRefGoogle Scholar
  16. 16.
    Guevara I, Iwanejko J, Dembinska-kiec A, Pankiewicz J, Wanat A, Anna P, lwona G, Bartus S, Malczewska-Malec M, Szczudlik A (1998) Determination of nitrite/nitrate in human biological material by the simple Griess reaction. Clin Chim Acta 274(2):177–188CrossRefGoogle Scholar
  17. 17.
    Rapp JP (1982) A genetic locus (Hyp-2) controlling vascular smooth muscle response in spontaneously hypertensive rats. Hypertension 4(4):459–467CrossRefGoogle Scholar
  18. 18.
    Kundu S, Shabir H, Basir SF, Khan LA (2014) Inhibition of As (III) and Hg (II) caused aortic hypercontraction by eugenol, linalool and carvone. J Smooth Muscle Res 50:93–102CrossRefGoogle Scholar
  19. 19.
    Leonard S, Gannett PM, Rojanasakul Y, Schwegler-Berry D, Castranova V, Vallyathan V, Shi X (1998) Cobalt-mediated generation of reactive oxygen species and its possible mechanism. J Inorg Biochem 70(3–4):239–244CrossRefGoogle Scholar
  20. 20.
    Chachami G, Simos G, Hatziefthimiou A, Bonanou S, Molyvdas P-A, Paraskeva E (2004) Cobalt induces hypoxia-inducible factor-1α expression in airway smooth muscle cells by a reactive oxygen species–and PI3K-dependent mechanism. Am J Respir Cell Mol Biol 31(5):544–551CrossRefGoogle Scholar
  21. 21.
    Taggart MJ, Wray S (1998) Hypoxia and smooth muscle function: key regulatory events during metabolic stress. J Physiol 509(2):315–325CrossRefGoogle Scholar
  22. 22.
    Shimizu S, Bowman PS, Thorne G, Paul RJ (2000) Effects of hypoxia on isometric force, intracellular Ca2+, pH, and energetics in porcine coronary artery. Circ Res 86(8):862–870CrossRefGoogle Scholar
  23. 23.
    Ahn B-H, Park MH, Lee YH, Kwon TK (2007) Up-regulation of cyclooxygenase-2 by cobalt chloride-induced hypoxia is mediated by phospholipase D isozymes in human astroglioma cells. Biochim Biophys Acta 1773(12):1721–1731CrossRefGoogle Scholar
  24. 24.
    Marletta MA (1993) Nitric oxide synthase structure and mechanism. J Biol Chem 268:12231–12234Google Scholar
  25. 25.
    Raman CS, Li H, Martasek P, Kral V, Masters BS, Poulos TL (1998) Crystal structure of constitutive endothelial nitric oxide synthase:a paradigm for pterin function involving a novel metal center. Cell 95:939–950CrossRefGoogle Scholar
  26. 26.
    Xia Y, Tsai AL, Berka V, Zweier JL (1998) Superoxide generation from endothelial nitric-oxide synthase. A Ca2+/calmodulin dependent and tetrahydrobiopterin regulatory process. J Biol Chem 273:25804–25808CrossRefGoogle Scholar
  27. 27.
    Moncada S, Rees DD, Schulz R, Palmer RM (1991) Development and mechanism of a specific supersensitivity to nitrovasodilators after inhibition of vascular nitric oxide synthesis in vivo. Proc Natl Acad Sci 88(6):2166–2170CrossRefGoogle Scholar
  28. 28.
    Gryglewski RJ, Palmer RMJ, Moncada S (1986) Superoxide anion is involved in the breakdown of endothelium-derived vascular relaxing factor. Nature 320(6061):454–456CrossRefGoogle Scholar
  29. 29.
    Macarthur H, Westfall TC, Wilken GH (2008) Oxidative stress attenuates NO-induced modulation of sympathetic neurotransmission in the mesenteric arterial bed of spontaneously hypertensive rats. Am J Phys Heart Circ Phys 294(1):H183–H189Google Scholar
  30. 30.
    Rubanyi GM, Vanhoutte PM (1986) Superoxide anions and hyperoxia inactivate endothelium-derived relaxing factor. Am J Phys Heart Circ Phys 250(5):H822–H827Google Scholar
  31. 31.
    Shimizu K, Kaneda T, Chihara H, Kaburagi T, Nakajyo S, Urakawa N (1995) Effects of phenylephrine on the contractile tension and cytosolic Ca2+ level in rat anococcygeus muscle. J Smooth Muscle Res 31:163–174CrossRefGoogle Scholar
  32. 32.
    Pillai S, Bikle DD (1992) Lanthanum influx into cultured human keratinocytes: effect on calcium flux and terminal differentiation. J Cell Physiol 151(3):623–629CrossRefGoogle Scholar
  33. 33.
    Ok S-H, Kwon S-C, Kang S, Choi M-J, Sohn J-T (2014) Mepivacaine-induced intracellular calcium increase appears to be mediated primarily by calcium influx in rat aorta without endothelium. Korean J Anesthesiol 67(6):404–411CrossRefGoogle Scholar
  34. 34.
    Caldwell RA, Clemo HF, Baumgarten CM (1998) Using gadolinium to identify stretch-activated channels: technical considerations. Am J Phys Cell Phys 275(2):C619–C621CrossRefGoogle Scholar
  35. 35.
    Wani SA, Khan LA, Basir SF (2018) Role of calcium channels and endothelial factors in nickel induced aortic hypercontraction in Wistar rats. J Smooth Muscle Res 54:71–82CrossRefGoogle Scholar
  36. 36.
    Tabet F, Savoia C, Schiffrin EL, Touyz RM (2004) Differential calcium regulation by hydrogen peroxide and superoxide in vascular smooth muscle cells from spontaneously hypertensive rats. J Cardiovasc Pharmacol 44(2):200–208CrossRefGoogle Scholar
  37. 37.
    Zimmerman MC, Takapoo M, Jagadeesha DK, Stanic B, Banfi B, Bhalla RC, Miller FJ Jr (2011) Activation of NADPH oxidase 1 increases intracellular calcium and migration of smooth muscle cells. Hypertension 58(3):446–453CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Shahnawaz Ahmad Wani
    • 1
  • Luqman Ahmad Khan
    • 1
  • Seemi Farhat Basir
    • 1
    Email author
  1. 1.Department of BiosciencesJamia Millia IslamiaNew DelhiIndia

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