Molecular and Cellular Biochemistry

, Volume 359, Issue 1–2, pp 409–418 | Cite as

Testosterone-dependent increase in blood pressure is mediated by elevated Cyp4A expression in fructose-fed rats

  • Harish Vasudevan
  • Violet G. Yuen
  • John H. McNeill


Endothelial dysfunction and increased blood pressure following insulin resistance play an important role in the development of secondary cardiovascular complications. The presence of testosterone is essential for the development of endothelial dysfunction and increased blood pressure. Testosterone regulates the synthesis of vasoconstrictor eicosanoids such as 20-hydroxyeicosatetranoic acid (20-HETE). In a series of studies, we examined: (1) the role of the androgen receptor in elevating blood pressure and (2) the effects of Cyp4A-catalyzed 20-HETE synthesis on vascular reactivity and blood pressure in fructose-fed rats. In the first study, intact and castrated male rats were made insulin resistant by feeding fructose for 9 weeks following which their superior mesenteric arteries (SMA) were isolated and examined for changes in endothelium-dependent relaxation in the presence and absence of 1-aminobenzotriazole (ABT) and N-methylsulfonyl-12,12-dibromododec-11-enamide (DDMS), which are inhibitors of 20-HETE synthesis. In another study, male rats were treated with either ABT or the androgen receptor blocker, flutamide, following which changes in insulin sensitivity, blood pressure, and vascular Cyp4A expression were measured. In the final study, HET0016, which is a more selective inhibitor of 20-HETE synthesis, was used to confirm our earlier findings. Treatment with HET0016 or ABT prevented or ameliorated the increase in blood pressure. Gonadectomy or flutamide prevented the increase in both the Cyp4A and blood pressure. Furthermore, both ABT and DDMS improved relaxation only in the intact fructose-fed rats. Taken together our results suggest that in the presence of testosterone, the Cyp4A/20-HETE system plays a key role in elevating the blood pressure secondary to insulin resistance.


Insulin resistance Blood pressure Testosterone Cyp4A 20-HETE 



This project was funded by grants-in aid from The Heart and Stroke Foundation of BC and Yukon (HSFBCY) and Canadian Institutes of Health Research (CIHR-Priority announcement grant from the Institute of Gender and Health) to Dr. McNeill. Harish Vasudevan was funded by a Doctoral Research Award from the Heart and Stroke Foundation of Canada and a Senior Graduate Studentship from the Michael Smith Foundation for Health Research British Columbia, Canada.


  1. 1.
    Vasudevan H, Nagareddy PR, McNeill JH (2006) Gonadectomy prevents endothelial dysfunction in fructose-fed male rats, a factor contributing to the development of hypertension. Am J Physiol Heart Circ Physiol 291:H3058–H3064PubMedCrossRefGoogle Scholar
  2. 2.
    Verma S, Bhanot S, Yao L, McNeill JH (1996) Defective endothelium-dependent relaxation in fructose-hypertensive rats. Am J Hypertens 9:370–376PubMedCrossRefGoogle Scholar
  3. 3.
    Katakam PV, Ujhelyi MR, Miller AW (1999) EDHF-mediated relaxation is impaired in fructose-fed rats. J Cardiovasc Pharmacol 34:461–467PubMedCrossRefGoogle Scholar
  4. 4.
    Song D, Arikawa E, Galipeau D, Battell M, McNeill JH (2004) Androgens are necessary for the development of fructose-induced hypertension. Hypertension 43:667–672PubMedCrossRefGoogle Scholar
  5. 5.
    Vasudevan H, Xiang H, McNeill JH (2005) Differential regulation of insulin resistance and hypertension by sex hormones in fructose-fed male rats. Am J Physiol Heart Circ Physiol 289:H1335–H1342PubMedCrossRefGoogle Scholar
  6. 6.
    Reckelhoff JF, Zhang H, Srivastava K, Granger JP (1999) Gender differences in hypertension in spontaneously hypertensive rats: role of androgens and androgen receptor. Hypertension 34:920–923PubMedGoogle Scholar
  7. 7.
    Reckelhoff JF, Zhang H, Srivastava K (2000) Gender differences in development of hypertension in spontaneously hypertensive rats: role of the renin-angiotensin system. Hypertension 35:480–483PubMedGoogle Scholar
  8. 8.
    Yanes LL, Sartori-Valinotti JC, Iliescu R, Romero DG, Racusen LC, Zhang H, Reckelhoff JF (2009) Testosterone-dependent hypertension and upregulation of intrarenal angiotensinogen in Dahl salt-sensitive rats. Am J Physiol Renal Physiol 296:F771–F779PubMedCrossRefGoogle Scholar
  9. 9.
    Bogatcheva NV, Sergeeva MG, Dudek SM, Verin AD (2005) Arachidonic acid cascade in endothelial pathobiology. Microvasc Res 69:107–127PubMedCrossRefGoogle Scholar
  10. 10.
    Vanhoutte PM, Feletou M, Taddei S (2005) Endothelium-dependent contractions in hypertension. Br J Pharmacol 144:449–458PubMedCrossRefGoogle Scholar
  11. 11.
    Galipeau D, Arikawa E, Sekirov I, McNeill JH (2001) Chronic thromboxane synthase inhibition prevents fructose-induced hypertension. Hypertension 38:872–876PubMedGoogle Scholar
  12. 12.
    Jiang J, Tran L, Vasudevan H, Xia Z, Yuen VG, McNeill JH (2007) Endothelin-1 blockade prevents COX2 induction and TXA2 production in the fructose hypertensive rat. Can J Physiol Pharmacol 85:422–429PubMedCrossRefGoogle Scholar
  13. 13.
    Vasudevan H, Lau SM, Jiang J, Galipeau D, McNeill JH (2010) Effects of insulin resistance and testosterone on the participation of cyclooxygenase isoforms in vascular reactivity. J Exp Pharmacol 2:169–179Google Scholar
  14. 14.
    Vasudevan H (2009) Testosterone-dependent vascular arachidonic acid metabolism in the regulation of insulin resistance and blood pressure. Faculty of Pharmaceutical Sciences, University of British Columbia, Vancouver, pp 1–151Google Scholar
  15. 15.
    Roman RJ (2002) P-450 metabolites of arachidonic acid in the control of cardiovascular function. Physiol Rev 82:131–185PubMedGoogle Scholar
  16. 16.
    Holla VR, Adas F, Imig JD, Zhao X, Price E Jr, Olsen N, Kovacs WJ, Magnuson MA, Keeney DS, Breyer MD, Falck JR, Waterman MR, Capdevila JH (2001) Alterations in the regulation of androgen-sensitive Cyp 4a monooxygenases cause hypertension. Proc Natl Acad Sci USA 98:5211–5216PubMedCrossRefGoogle Scholar
  17. 17.
    Nakagawa K, Marji JS, Schwartzman ML, Waterman MR, Capdevila JH (2003) Androgen-mediated induction of the kidney arachidonate hydroxylases is associated with the development of hypertension. Am J Physiol Regul Integr Comp Physiol 284:R1055–R1062PubMedGoogle Scholar
  18. 18.
    Singh H, Cheng J, Deng H, Kemp R, Ishizuka T, Nasjletti A, Schwartzman ML (2007) Vascular cytochrome P450 4A expression and 20-hydroxyeicosatetraenoic acid synthesis contribute to endothelial dysfunction in androgen-induced hypertension. Hypertension 50:123–129PubMedCrossRefGoogle Scholar
  19. 19.
    Miyata N, Roman RJ (2005) Role of 20-hydroxyeicosatetraenoic acid (20-HETE) in vascular system. J Smooth Muscle Res 41:175–193PubMedCrossRefGoogle Scholar
  20. 20.
    Cheng J, Ou JS, Singh H, Falck JR, Narsimhaswamy D, Pritchard KA Jr, Schwartzman ML (2008) 20-Hydroxyeicosatetraenoic acid causes endothelial dysfunction via eNOS uncoupling. Am J Physiol Heart Circ Physiol 294:H1018–H1026PubMedCrossRefGoogle Scholar
  21. 21.
    Cheng J, Ou JS, Singh H, Falck JR, Narsimhaswamy D, Pritchard KA Jr, Schwartzman ML (2008) 20-Hydroxyeicosatetraenoic acid causes endothelial dysfunction via eNOS uncoupling. American journal of physiology. Heart and circulatory physiology 294:H1018–H1026PubMedCrossRefGoogle Scholar
  22. 22.
    Cheng J, Wu CC, Gotlinger KH, Zhang F, Falck JR, Narsimhaswamy D, Schwartzman ML (2010) 20-hydroxy-5,8,11,14-eicosatetraenoic acid mediates endothelial dysfunction via IkappaB kinase-dependent endothelial nitric-oxide synthase uncoupling. J Pharmacol Exp Ther 332:57–65PubMedCrossRefGoogle Scholar
  23. 23.
    Berezan DJ, Dunn KM, Falck JR, Davidge ST (2008) Aging increases cytochrome P450 4A modulation of alpha1-adrenergic vasoconstriction in mesenteric arteries. J Cardiovasc Pharmacol 51:327–330PubMedCrossRefGoogle Scholar
  24. 24.
    Croft KD, McGiff JC, Sanchez-Mendoza A, Carroll MA (2000) Angiotensin II releases 20-HETE from rat renal microvessels. Am J Physiol Renal Physiol 279:F544–F551PubMedGoogle Scholar
  25. 25.
    Barnett CR, Rudd S, Flatt PR, Ioannides C (1993) Sex differences in the diabetes-induced modulation of rat hepatic cytochrome P450 proteins. Biochem Pharmacol 45:313–319PubMedCrossRefGoogle Scholar
  26. 26.
    Benter IF, Yousif MH, Canatan H, Akhtar S (2005) Inhibition of Ca2+/calmodulin-dependent protein kinase II, RAS-GTPase and 20-hydroxyeicosatetraenoic acid attenuates the development of diabetes-induced vascular dysfunction in the rat carotid artery. Pharmacol Res 52:252–257PubMedCrossRefGoogle Scholar
  27. 27.
    Chen YJ, Li J, Quilley J (2008) Deficient renal 20-HETE release in the diabetic rat is not the result of oxidative stress. Am J Physiol Heart Circ Physiol 294:H2305–H2312PubMedCrossRefGoogle Scholar
  28. 28.
    Laffer CL, Laniado-Schwartzman M, Nasjletti A, Elijovich F (2004) 20-HETE and circulating insulin in essential hypertension with obesity. Hypertension 43:388–392PubMedCrossRefGoogle Scholar
  29. 29.
    Wang MH, Smith A, Zhou Y, Chang HH, Lin S, Zhao X, Imig JD, Dorrance AM (2003) Downregulation of renal CYP-derived eicosanoid synthesis in rats with diet-induced hypertension. Hypertension 42:594–599PubMedCrossRefGoogle Scholar
  30. 30.
    Tsai IJ, Croft KD, Mori TA, Falck JR, Beilin LJ, Puddey IB, Barden AE (2009) 20-HETE and F2-isoprostanes in the metabolic syndrome: the effect of weight reduction. Free Radic Biol Med 46:263–270PubMedCrossRefGoogle Scholar
  31. 31.
    Ward NC, Hodgson JM, Puddey IB, Beilin LJ, Croft KD (2006) 20-Hydroxyeicosatetraenoic acid is not associated with circulating insulin in lean to overweight humans. Diabetes Res Clin Pract 74:197–200PubMedCrossRefGoogle Scholar
  32. 32.
    Dunn KM, Renic M, Flasch AK, Harder DR, Falck J, Roman RJ (2008) Elevated production of 20-HETE in the cerebral vasculature contributes to severity of ischemic stroke and oxidative stress in spontaneously hypertensive rats. Am J Physiol Heart Circ Physiol 295:H2455–H2465PubMedCrossRefGoogle Scholar
  33. 33.
    Hwang IS, Ho H, Hoffman BB, Reaven GM (1987) Fructose-induced insulin resistance and hypertension in rats. Hypertension 10:512–516PubMedGoogle Scholar
  34. 34.
    Llinas MT, Alexander BT, Capparelli MF, Carroll MA, Granger JP (2004) Cytochrome P-450 inhibition attenuates hypertension induced by reductions in uterine perfusion pressure in pregnant rats. Hypertension 43:623–628PubMedCrossRefGoogle Scholar
  35. 35.
    Miyata N, Taniguchi K, Seki T, Ishimoto T, Sato-Watanabe M, Yasuda Y, Doi M, Kametani S, Tomishima Y, Ueki T, Sato M, Kameo K (2001) HET0016, a potent and selective inhibitor of 20-HETE synthesizing enzyme. Br J Pharmacol 133:325–329PubMedCrossRefGoogle Scholar
  36. 36.
    Sato M, Ishii T, Kobayashi-Matsunaga Y, Amada H, Taniguchi K, Miyata N, Kameo K (2001) Discovery of a N′-hydroxyphenylformamidine derivative HET0016 as a potent and selective 20-HETE synthase inhibitor. Bioorg Med Chem Lett 11:2993–2995PubMedCrossRefGoogle Scholar
  37. 37.
    Seki T, Wang MH, Miyata N, Laniado-Schwartzman M (2005) Cytochrome P450 4A isoform inhibitory profile of N-hydroxy-N′-(4-butyl-2-methylphenyl)-formamidine (HET0016), a selective inhibitor of 20-HETE synthesis. Biol Pharm Bull 28:1651–1654PubMedCrossRefGoogle Scholar
  38. 38.
    Hoagland KM, Flasch AK, Roman RJ (2003) Inhibitors of 20-HETE formation promote salt-sensitive hypertension in rats. Hypertension 42:669–673PubMedCrossRefGoogle Scholar
  39. 39.
    Blanton A, Nsaif R, Hercule H, Oyekan A (2006) Nitric oxide/cytochrome P450 interactions in cyclosporin A-induced effects in the rat. J Hypertens 24:1865–1872PubMedCrossRefGoogle Scholar
  40. 40.
    Vasudevan H (2005) Potential role of sex hormones in altered vascular relaxation following insulin resistance. Faculty of Pharmaceutical Sciences, University of British Columbia, Vancouver, pp 1–99Google Scholar
  41. 41.
    Matsuda M, DeFronzo RA (1999) Insulin sensitivity indices obtained from oral glucose tolerance testing: comparison with the euglycemic insulin clamp. Diabetes Care 22:1462–1470PubMedCrossRefGoogle Scholar
  42. 42.
    Ito O, Alonso-Galicia M, Hopp KA, Roman RJ (1998) Localization of cytochrome P-450 4A isoforms along the rat nephron. Am J Physiol 274:F395–F404PubMedGoogle Scholar
  43. 43.
    Katakam PV, Ujhelyi MR, Hoenig ME, Miller AW (1998) Endothelial dysfunction precedes hypertension in diet-induced insulin resistance. Am J Physiol 275:R788–R792PubMedGoogle Scholar
  44. 44.
    Linder CD, Renaud NA, Hutzler JM (2009) Is 1-aminobenzotriazole an appropriate in vitro tool as a nonspecific cytochrome P450 inactivator? Drug Metab Dispos 37:10–13PubMedCrossRefGoogle Scholar
  45. 45.
    Hong HJ, Liu JC, Chan P, Juan SH, Loh SH, Lin JG, Cheng TH (2004) 17b-estradiol downregulates angiotensin-II-induced endothelin-1 gene expression in rat aortic smooth muscle cells. J Biomed Sci 11:27–36PubMedGoogle Scholar
  46. 46.
    Nagareddy PR (2009) Mechanisms of vascular dysfunction in diabetes and hypertension. Faculty of Pharmaceutical Sciences, University of British Columbia, Vancouver, p 267Google Scholar

Copyright information

© Springer Science+Business Media, LLC. 2011

Authors and Affiliations

  • Harish Vasudevan
    • 1
  • Violet G. Yuen
    • 1
  • John H. McNeill
    • 1
  1. 1.Division of Pharmacology and Toxicology, Faculty of Pharmaceutical SciencesUniversity of British ColumbiaVancouverCanada

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