Skip to main content
Log in

Exercise conditioning attenuates the hypertensive effects of nitric oxide synthase inhibitor in rat

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

Many individuals with cardiovascular diseases undergo periodic exercise conditioning with or with out medication. Therefore, this study investigated the interaction of exercise training and chronic nitric oxide synthase (NOS) inhibitor (Nitro-L-Arginine Methyl Ester, L-NAME) treatment on blood pressure and its correlation with aortic nitric oxide (NO), antioxidant defense system and oxidative stress parameters in rats. Fisher 344 rats were divided into four groups: (1) sedentary control, (2) exercise training (ET) for 8 weeks, (3) L-NAME (10 mg/kg, subcutaneous for 8 weeks) and (4) ET + L-NAME. Blood pressure (BP) was monitored weekly for 8 weeks with tail-cuff method. The animals were sacrificed 24 h after last treatments and thoracic aortic rings were isolated and analyzed. Exercise conditioning resulted in a significant increase in respiratory exchange ratio (RER), aortic NO production, NO synthase activity and inducible iNOS protein expression. Training significantly enhanced aortic GSH levels, GSH/GSSG ratio and up-regulation of aortic CuZn-SOD, Mn-SOD, catalase (CAT) glutathione peroxidase (GSH-Px) activity and protein expression and significantly decreased aortic lipid peroxidation. Chronic L-NAME administration resulted in a significant depletion of aortic NO, NOS activity, endothelial (eNOS) and iNOS protein expression, GSH level, GSH/GSSG ratio, down-regulation of aortic antioxidant enzyme activities and protein expressions. Aortic xanthine oxidase (XO) activity significantly increased with increased lipid peroxidation and protein oxidation after L-NAME administration. The biochemical changes were accompanied by increased in BP. Interaction of training and chronic NOS inhibitor treatment resulted in normalization of BP and aortic antioxidant enzyme activity and protein expression, up-regulation of aortic GSH/GSSG ratio, NO levels, Mn-SOD protein expression, depletion of GSSG, protein oxidation and lipid peroxidation. The data suggest that training attenuated the oxidative injury caused by chronic NOS inhibitor treatment by up-regulating the NO and antioxidant systems and lowering the BP in rats.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Belch JJF, Bridges AB, Scott N, Chopra M: Oxygen free radicals and congestive heart failure. Br Heart J 65: 245–248, 1991

    Google Scholar 

  2. Somani SM, Husain K: Influence of exercise induced oxidative stress on the central nervous system. In: C.K. Sen, L. Packer, O. Hanninen (eds). Hand Book of Oxidants and Antioxidants in Exercise, 2nd edn. Elsevier Science B.V., Amsterdam, 2000, pp 713–754

    Google Scholar 

  3. Das UN: Free radicals, cytokines and nitric oxide in cardiac failure and myocardial infarction. Mol Cell Biochem 215: 145–152, 2000

    Google Scholar 

  4. Sagar S, Kallo, IJ, Kaul N, Ganguly NK, Sharma BK: Oxygen free radicals in essential hypertension. Mol Cell Biochem 111: 103–108, 1992

    Google Scholar 

  5. Davies JJA, Qintanilha AT, Brooks GA, Packer L: Free radicals and tissue damage produced by exercise. Biochem Biophys Res Commun 107: 1198–1205, 1982

    Google Scholar 

  6. Powers SK, Criswell D, Lawler J, Marti, D, Lieu F, Ji LL, Herb RA: Rigorous exercise training increases superoxide dismutase activity in ventricular myocardium. Am J Physiol 265: 2094–2098, 1993

    Google Scholar 

  7. Somani SM, Rybak LP, Frank S: Effect of acute and trained exercise on antioxidant system in rat heart subcellular fraction. Pharmacol Biochem Behav 51: 627–634, 1995

    Google Scholar 

  8. Somani SM, Husain K: Exercise training alters kinetics of antioxidant enzymes in rat tissues. Biochem Mol Biol Int 38: 587–595, 1996

    Google Scholar 

  9. Husain K, Somani SM: Response of exercise training and chronic ethanol ingestion on cardiac antioxidant system of rat. Alcohol 14: 301–307, 1997

    Google Scholar 

  10. Sessa WC, Pritchard K, Syedi N, Wang J, Hintze TH: Chronic exercise in dogs increases coronary vascular nitric oxide production and endothelial cell nitric oxide synthase gene expression. Circ Res 74: 349–353, 1993

    Google Scholar 

  11. Wang J, Wolin MS, Hintze TH: Chronic exercise enhances endothelium-mediated dilation of epicardial coronary artery in conscious dogs. Circ Res 73: 829–838, 1993

    Google Scholar 

  12. Humphrey R, Bartels M N: Exercise, cardiovascular disease and chronic heart failure. Arch Phys Med Rehab 82: S76–S81, 2001

    Google Scholar 

  13. Pace B: Benefits of physical activity for the heart. J Am Med Assoc 285: 1536, 2001

    Google Scholar 

  14. Yang Z, Zheng T, Zhang A, Altura BT, Altura BM: Mechanisms of hydrogen peroxide induced contraction of rat aorta. Eur J Pharmacol 344: 169–181, 1998

    Google Scholar 

  15. Stauss HM, Nafz B, Mrowka R, Persson PB: Blood pressure control in eNOS knock out mice: Comparison with other species under NO blockade. Acta Physiol Scand 168: 155–160, 2000

    Google Scholar 

  16. Lounsbury KM, Hu Q, Ziegelstein RC: Calcium signaling and oxidant stress in the vasculature. Free Rad Biol Med 28: 1362–1369, 2000

    Google Scholar 

  17. Rubanyi G M, Ho EH, Cantor EH, Inmma WC, Parker B: Cytoprotective function of nitric oxide: Inactivation of superoxide radicals produced by human leukocytes. Biochem Biophys Res Commun 181: 1392–1397, 1991

    Google Scholar 

  18. Beckman JS, Beckman TW, Chen J, Marshall PA, Freeman BA: Apparent hydroxyl radical production by peroxinitrite: Implications for endothelial injury from nitric oxide and superoxide. Proc Natl Acad Sci USA 87: 1620–1624, 1990

    Google Scholar 

  19. Gardiner SM, Compton AM, Bennett T, Palmer RJJ, Moncada S: Regional hemodynamic changes during oral ingestion of N-nitro-L-arginine methyl ester in conscious Brattelboro rats. Br J Pharmacol 101: 10–12, 1990

    Google Scholar 

  20. Bank N, Aynedjian HS, Khan GA: Mechanism of vasoconstriction induced by chronic inhibition of nitric oxide in rats. Hypertension 24: 322–328, 1994

    Google Scholar 

  21. Tribulova N, Okruhlicova L, Bernatova I, Pechanova O: Chronic disturbances in NO production results in histochemical and subcellular alterations of the rat heart. Physiol Res 49: 77–88, 2000

    Google Scholar 

  22. Hably C, Vag G, Bartha J: Nitric oxide synthase inhibition increases vascular resistance in sodium and water loaded rats. Res Exp Med (Berl) 198: 145–156, 1998

    Google Scholar 

  23. Jolma P, Kalliovalkama J, Tolvanen JP, Koobi P, Kahonen M, Hurti-Kahonen N, Wu X, Porsti I: High calcium diet enhances vasorelaxation in nitric oxide-deficient hypertension. Am J Physiol Heart Circ Physiol 279: H1036–H1043, 2000

    Google Scholar 

  24. Fariss WM, Reed DJ: High performance liquid chromatography of thiols and disulfides: Dinitrophenol derivatives. Meth Enzymol 143: 101–109, 1987

    Google Scholar 

  25. Misra H P, Fridovich I: The role of superoxide anion in the autoxidation of epinephrine and a simple assay for superoxide dismutase. J Biol Chem 243: 3170–3175, 1972

    Google Scholar 

  26. Aebi H: Catalase. Meth Enzymol 105: 125–126, 1984

    Google Scholar 

  27. Flohe L, Gunzler WA: Glutathione peroxidase. Meth Enzymol 105: 115–121, 1984

    Google Scholar 

  28. Carlberg L, Mannervik B: Glutathione reductase. Meth Enzymol 113: 484–499, 1985

    Google Scholar 

  29. Habig WH, Pabst MJ, Jakoby WB: Glutathione-s-transferases. The first step in mercapturic acid formation. J Biol Chem 249: 7130–7139, 1974

    Google Scholar 

  30. Singh NM, Bogavac L, Kalimanovska V, Jelic Z, Spasic S: Spectrophotometric assay of xanthine oxidase with 2,2-azino-di(3-methylbenzthiazoline-6-sulphonate) (ABST) as chromogen. Clin Chem Acta 162: 29–36, 1987

    Google Scholar 

  31. Husain K: Interaction of exercise training and chronic NOS inhibition on blood pressure, heart rate, NO and antioxidants in plasma of rats. Pathophysiology 2001 (under review)

  32. Ohkawa H, Ohishi N, Yagi K: Assay for lipid peroxides in animal and tissues by thiobarbituric acid reaction. Anal Biochem 95: 351–358, 1979

    Google Scholar 

  33. Buss H, Chan TP, Sluis KB, Domigan NM, Winterbourn CC: Protein carbonyl measurement by a sensitive ELISA method. Free Rad Biol Med 23: 361–366, 1997

    Google Scholar 

  34. Read SM, Northcole DH: Minimization of variation in the response to different protein of the Coomassie blue G-dye-binding assay for protein. Anal Biochem 116: 53–64, 1981

    Google Scholar 

  35. Chen SJ, Wu CC, Yen MH: Exercise training activates large-conductance calcium activated K(+) channels and enhances nitric oxide production in rat mesentric artery and thoracic aorta. J Biomed Sci 8: 248–255, 2001

    Google Scholar 

  36. Brum PC, DaSilva GJ, Moreira ED, Ida F, Negrao CE, Krieger EM: Exercise training increases baroreceptor gain sensitivity in normal and hypertensive rats. Hypertension 36: 1018–1022, 2000

    Google Scholar 

  37. Spier SA, Laughlin MH, Delp MD: Effects of acute and chronic exercise on vasoconstrictor responsiveness of rat abdominal aorta. J Appl Physiol 87: 1752–1757, 1999

    Google Scholar 

  38. Levy WC, Cerqueira MD, Harp GD, Johannessen K, Abrass IB, Schwartz RS, Stratton JR: Effect of endurance exercise training on heart rate variability at rest in healthy young and older men. Am J Cardiol 82: 1236–1241, 1998

    Google Scholar 

  39. Versa-Silva AS, Mattos KC, Gava NS, Brum PC, Negrao CE, Krieger EM: Low-intensity exercise training decreases cardiac output and hypertension in spontaneously hypertensive rats. Am J Physiol 273: H2627–H2631, 1997

    Google Scholar 

  40. Chen HI, Cheng SY, Jen CJ: Chronic exercise enhances vascular responses to clonidine in rats by increasing endothelial alpha2-adrenergic receptor affinity. Chin J Physiol 42: 61–66, 1999

    Google Scholar 

  41. Chen HI, Li HT, Chen CC: Physical conditioning decreases norepinephrine-induced vasoconstriction in rabbits. Possible roles of norepinephrine-evoked endothelium-derived relaxing facto. Circulation 90: 970–975, 1994

    Google Scholar 

  42. Balligand JL, Ungureanu-Longrois D, Simmons WW, Pimental D, Malinski TA, Kapturczak M, Taha Z, Lowenstein CJ, Davidoff AJ, Kelly RA, Smith TW, Michel T: Cytokine-inducible nitric oxide synthase (iNOS) expression in cardiac myocytes. J Biol Chem 279: 27580–27588, 1994

    Google Scholar 

  43. Balligand JL, Kobzik L, Han X, Kaye DM, Belhassen L, OHara DS, Kelly RA, Smith TW, Michel T: Nitric oxide dependent parasympathetic signaling is due to activation of constitutive endothelial (type III) nitric oxide synthase in cardiac myocytes. J Biol Chem 270: 14582–14588, 1995

    Google Scholar 

  44. Fukai T, Siegfried MR, Ushio-Fukai M, Cheng Y, Kojda G, Harrison DG: Regulation of the vascular extracellular superoxide dismurase by nitric oxide and exercise training. J Clin Invest 105: 1631–1639, 2000

    Google Scholar 

  45. Iemitsu M, Miyauchi T, Maeda S, Yuki K, Kobayashi T, Kumagai Y, Shimojo N, Yamaguchi I, Matsuda M: Intense exercise causes decrease in expression of both endothelial NO synthase and tissue Nox levels in hearts. Am J Physiol Reg Integr Comp Physiol 279: R951–R959, 2000

    Google Scholar 

  46. Gyurko R, Kuhlencordt P, Fishman MC, Huang PL: Modulation of mouse cardiac function in vivo by eNOS and ANP. Am J Physiol Heart Circ Physiol 278: H971–H981, 2000

    Google Scholar 

  47. Linke A, Shoene N, Gielen S, Hofer J, Herbs S, Schuler G, Hambrecht R: Endothelial dysfunction in patients with chronic heart failure: Systemic effects of lower-limb exercise training. J Am Coll Cardiol 37: 392–397, 2001

    Google Scholar 

  48. Joyner MJ, Shastry S: Vascular endothelial growth factor and capillary density in exercise training. Exer Sports Sci Rev 91: 97–98, 2000

    Google Scholar 

  49. Benoit H, Jordan M, Wagner H, Wagner PD: Effect of NO, vasodilator prostaglandins, and adenosine on skeletal muscle angeogenic growth factor gene expression. J Appl Physiol 86: 1513–1518, 1999

    Google Scholar 

  50. Sen CK: Glutathione homeostasis in response to exercise training and nutritional supplements. Mol Cell Biochem 196: 31–42, 1999

    Google Scholar 

  51. Powers SK, Ji LL, Leeuwenburgh C: Exercise training-induced alterations in skeletal muscle antioxidant capacity: A brief review. Med Sci Sports Exer 31: 987–997, 1999

    Google Scholar 

  52. Hong H, Johnson P: Antioxidant enzyme activities and lipid peroxidation levels in exercised and hypertensive rat tissues. Int J Biochem Cell Biol 27: 923–931, 1995

    Google Scholar 

  53. Demirel, HA, Powers SK, Caillaud C, Coombes JS, Naito H, Fletcher LA, Vrabas I, Jessup JV, Ji LL: Exercise training reduces myocardial lipid peroxidation following short-term ischemia-reperfusion. Med Sci Sports Exer 30: 1211–1226, 1998

    Google Scholar 

  54. d'Ischia M, Palumbo A, Buzzo F: Interaction of nitric oxide with lipid peroxidation products under aerobic conditions: Inhibitory effects of the formation of malondialdehyde and related thiobarbituric acid -reactive substances. Nitric Oxide 4: 4–14, 2000

    Google Scholar 

  55. Somani SM, Rybak LP: Comparative effects of exercise training on transcription of antioxidant enzymes and the activity in old rat heart. Ind J Physiol Pharmacol 40: 205–212, 1996

    Google Scholar 

  56. Yamashita N, Hoshida S, Otsu K, Asahi M, Kuzuya T, Hori, M: Exercise provides direct biphasic cardioprotection via manganese superoxide dismutase activation. J Exp Med 189: 1699–1706, 1999

    Google Scholar 

  57. Fridovich I: Superoxide dismutases. Ann Rev Biochem 44: 147–159, 1975

    Google Scholar 

  58. Perera CS, St Clair DK, McClain CJ: Differential regulation of manganese superoxide dismutase activity by alcohol and TNF in human hepatoma cells. Arch Biochem Biophys 323: 471–476, 1995

    Google Scholar 

  59. Pigeolet E, Corbisier A, Houbion D, Lambert C, Michiel M, Raes M, Zachary D, Ramacle J: Glutathione peroxidase, superoxide dismutase and catalase inactivation by peroxides and oxygen derived radicals. Mech Age Dev 51: 283–2297, 1990

    Google Scholar 

  60. Suzuki K, Tatsumi H, Satoh S, Senda T, Nakata T, Fuji J, Taniguchi N: Manganese superoxide dismutase in endothelial cells: Localization and mechanism of induction. Am J Physiol 265: 1173–1178, 1993

    Google Scholar 

  61. Watkinson WP, Foley DH, Rubio R, Berne RM: Myocardial adenosine formation with increased cardiac performance in the dog. Am J Physiol 236: 13–21, 1979

    Google Scholar 

  62. Maggirwar SB, Dhanraj DN, Somani, SM, Ramkumar V. Adenosine acts as an endogenous activator of the cellular antioxidant defense system. Biochem Biophys Res Commun 201: 508–515, 1994

    Google Scholar 

  63. Meyer M, Pahl HL, Baeuerle PA: Regulation of the transcription factor NF-kappa B and AP-1 by redox changes. Chem Biol 91: 91–100, 1994

    Google Scholar 

  64. Sen CK, Packer L: Antioxidant and redox regulation of gene transcription. FASEB J 10: 709–720, 1996

    Google Scholar 

  65. Rubino A, Yellon DM: Ischaemic preconditioning of the vasculature: An overlooked phenomenon for protecting the heart? Trends Pharmacol Sci 21: 225–230, 2000

    Google Scholar 

  66. Qiu C, Muchant D, Beierwaltes WH, Racusen L, Baylis C: Evaluation of chronic nitric oxide inhibition hypertension: Relationship to renal function. Hypertension 31: 21–26, 1998

    Google Scholar 

  67. Golikov PP, Davydov BV, Merchenk VV, Nicolaeva N I, Golikov AP, Riabinin VA, Semenova EV, Polumiskov VI: Urapidil effects on oxidative stress in hypertensive crises. Klin Med (Mosk) 78: 42–45, 2000

    Google Scholar 

  68. Tomita H, Egashira K, Kubo-Inoue M, Usui M, Koyanagi M, Shimokawa H, Takeya, M, Yoshimura T, Takeshita A: Inhibition of NO synthesis induces inflammatory changes and monocyte chemoattractant protein-1 expression in rat hearts and vessels. Arterioscler Thromb Vasc Biol 18: 1456–1464, 1998

    Google Scholar 

  69. Nemeth I, Orvos H, Boda D: Blood glutathione redox status in gestational hypertension. Free Rad Biol Med 30: 715–721, 2001

    Google Scholar 

  70. Vaziri ND, Wang XQ, Oveisi F, Rad B: Induction of oxidative stress by glutathione depletion causes severe hypertension in normal rats. Hypertension 36: 142–146, 2000

    Google Scholar 

  71. Murakami E, Ishii J, Hiwada K, Kokubu T: The role of hypothalamic glutathione in hypertensive animals. Clin Exp Hypertens A 10: 347–352, 1988

    Google Scholar 

  72. Mihailovic MB, Avramovic DM, Jovanovic IB, Pesut OJ, Matic DP, Stojanov VJ: Blood and plasma selenium levels and GSH-Px activities in patients with arterial hypertension and chronic heart disease. J Environ Pathol Toxicol Oncol 17: 285–289, 1998

    Google Scholar 

  73. LaPier TLK, Rodnick KJ: Effects of aerobic exercise on energy metabolism in the hypertensive rat heart. Physical Ther 81: 1006–1017, 2001

    Google Scholar 

  74. Werner P, Cohen G: Glutathione disulfide (GSSG) as a marker of oxidative injury to brain mitochondria. Ann NY Acad Sci 679: 364–369, 1993

    Google Scholar 

Download references

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Husain, K. Exercise conditioning attenuates the hypertensive effects of nitric oxide synthase inhibitor in rat. Mol Cell Biochem 231, 129–137 (2002). https://doi.org/10.1023/A:1014416915643

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1014416915643

Navigation