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Hormetic acute response and chronic effect of ethanol on adenine nucleotide hydrolysis in rat platelets

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Abstract

The objective of this study was to verify the acute and chronic effects of ethanol on platelet NTPDase and 5′-nucleotidase activities. These enzymes modulate platelet function by regulating adenine nucleotide bioavailability and adenosine production. In the acute treatment, doses of 0.8, 2.0, 4.0, 6.0 and 8.0 g/kg ethanol were administered via orogastric tube, and induced a biphasic or hormetic effect on ATP, ADP and AMP platelet hydrolysis. Ethanol at a dose of 0.8 and 2.0 g/kg increased NTPDase activity (44 and 35%, P < 0.0001) with ATP as substrate, whereas when ADP was used there was only a tendency for NTPDase activity to increase. ATP and ADP hydrolysis decreased by 31–77% (P < 0.0001) in 4.0, 6.0 and 8.0 g/kg of ethanol compared to the control. AMP hydrolysis showed a tendency to increase at ethanol doses of 0.8 and 2.0 g/kg, but was inhibited by 45–100% (P < 0.0001) at the higher doses. Chronic treatment consisted of the oral administration of 20% ethanol solution during 31 weeks as the only source of liquid and inhibited NTPDase activity (15 and 20%, P < 0.05) with ATP and ADP as substrate, respectively. However, AMP hydrolysis by 5′-nucleotidase increased by 40% (P < 0.05). Thus, we speculate that the effects of ethanol on NTPDase and 5′-nucleotidase activities could be related with the platelets alterations commonly observed in alcohol users.

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References

  • Atkinson B, Dwyer K, Enjyoji K et al (2006) Ecto-nucleotidases of the CD39/NTPDase family and thrombus formation: potential as therapeutic targets. Blood Cells Mol Dis 36:217–222

    Article  PubMed  CAS  Google Scholar 

  • Baer D, Judd J, Clevidence B et al (2002) Moderate alcohol consumption lowers risk factors for cardiovascular disease in postmenopausal women fed a controlled diet. Am J Clin Nutr 75:593–599

    PubMed  CAS  Google Scholar 

  • Bakker WW, Poelstra K, Barradas MA et al (1994) Platelets and ectonucleotidases. Platelets 5:121–129

    Article  CAS  Google Scholar 

  • Beaugé F, Fleuret C, Barin F et al (1984) Brain membrane disordering after acute in vivo administration of ethanol, isopropanol or t-butanol in rats. Biochem Pharmacol 33:3591–3595

    Article  PubMed  Google Scholar 

  • Benaim G, Cervino V, Lopez-Estraño C et al (1994) Ethanol stimulates the plasma membrane calcium pump from human erythrocytes. Biochim Biophys Acta 1195:141–148

    Article  PubMed  CAS  Google Scholar 

  • Bergamini C, Grazi E (1980) Human platelets 5′-nucleotidase: a cell membrane ectoenzyme with a possible regulatory role in the aggregation reaction. Ital J Biochem 29:273–288

    PubMed  CAS  Google Scholar 

  • Birk AV, Broekman J, Gladek EM et al (2002) Role of extracellular ATP metabolism in regulation of platelet reactivity. J Lab Clin Med 140:166–175

    Article  PubMed  CAS  Google Scholar 

  • Blockland A, Prickaerts J, Raajmakers W (1992) Reduced level of anxiety in adult Lewis rats after chronic ethanol consumption. Physiol Behav 51:245–248

    Article  Google Scholar 

  • Boarder MR, Hourani SMO (1998) The regulation of vascular function by P2 receptors: multiple sites and multiple receptors. TiPS 19:99–107

    PubMed  CAS  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:218–254

    Article  Google Scholar 

  • Calabrese EJ (2002) Hormesis: changing view of the dose-response, a personal account of the history and current status. Mutat Res 511:181–189

    Article  PubMed  CAS  Google Scholar 

  • Calabrese EJ (2005) Paradigm lost, paradigm found: the re-emergence of hormesis as a fundamental dose response model in the toxicological sciences. Environ Pollut 138:378–411

    Article  CAS  Google Scholar 

  • Calabrese EJ, Baldwin LA (2003) Ethanol and hormesis. Critic Rev Toxicol 33:407–424

    Article  CAS  Google Scholar 

  • Carrasco MP, Jimenez-Lopez JM, Segovia JL et al (2007) Effects of ethanol on the remodeling of neutral lipids and phospholipids in brain mitochondria and microsomes. Neurochem Int 50:858–865

    Article  PubMed  CAS  Google Scholar 

  • Casamenti F, Scali C, Vannucchi MG, Bartolini L, Pepeu G (1993) Long-term ethanol consumption by rats: effect on acetylcholine release in vivo, choline acetyltransferase activity, and behavior. Neuroscience 56:465–471

    Article  PubMed  CAS  Google Scholar 

  • Chan K, Delfert D, Junger KD (1986) A direct colorimetric assay for Ca2+-ATPase activity. Anal Biochem 157:375–380

    Article  PubMed  CAS  Google Scholar 

  • Coade SB, Pearson JD (1989) Metabolism of adenine nucleotides in human blood. Circulat Res 65:531–537

    PubMed  CAS  Google Scholar 

  • Collins AC, Smolen A, Wayman AL, Marks MJ (1984) Ethanol and temperature effects on five membrane bound enzymes. Alcohol 1:237–246

    Article  PubMed  CAS  Google Scholar 

  • Côté YP, Picher M, St-Jean P et al (1991) Identification and localization of ATP-diphosphohydrolase (apyrase) in bovine aorta: relevance to vascular tone and platelet aggregation. Biochim Biophy Acta 1078:187–191

    Google Scholar 

  • Cristalli G, Volpini R, Vittori S (1994) 2-alkynil derivatives of adenosine-5′-N-ethyluronamide: selective A2 adenosine receptor agonists with potent inhibitory activity on platelet aggregation. J Med Chem 37:1720–1726

    Article  PubMed  CAS  Google Scholar 

  • Di Virgilio F, Chiozzi P, Ferrari D et al (2001) Nucleotide receptors: an emerging family of regulatory molecules in blood cells. Blood 97:587–600

    Article  PubMed  CAS  Google Scholar 

  • Drosopoulos JHF (2002) Roles as Asp54 and Asp213 in Ca2+ utilization by soluble human CD39/ecto-nucleotidase. Arch Biochem Bioph 406:85–95

    Article  CAS  Google Scholar 

  • Duarte AP, Dong QS, Young J et al (1995) Inhibition of platelet aggregation in whole blood by ethanol. Thromb Res 78:107–115

    Article  Google Scholar 

  • Foley TD, Rhoads DE (1992) Effects of ethanol on Na+-dependent amino acid uptake: dependence on rat age and Na+, K+-ATPase activity. Brain Res 594:39–43

    Article  Google Scholar 

  • Foster DM, Huber MD, Klemm WR (1989) Ethanol may stimulate or inhibit (Na+, K+) ATPase, depending upon Na+ and K+ concentrations. Alcohol 6:437–443

    Article  PubMed  CAS  Google Scholar 

  • Frasseto SS, Dias RD, Sarkis JJF (1993) Characterization of an ATP diphosphohydrolase activity (apyrase, EC 3.6.1.5) in rat blood platelets. Mol Cel Biochem 129:47–55

    Article  Google Scholar 

  • Frasseto SS, Dias RD, Sarkis JJF (1995) Inhibition and kinetic alterations by excess free ATP and ADP of the ATP diphosphohydrolase activity (EC 3.6.1.5) from rat blood platelets. Biochem Mol Biol Intern 35:499–506

    Google Scholar 

  • Fuchs CS, Stampfer MJ, Colditz GA (1995) Alcohol consumption and mortality among women. New Eng J Med 332:1245–1250

    Article  PubMed  CAS  Google Scholar 

  • Gangadharan SP, Imai M, Rhynhart KK et al (2001) Targeting platelet aggregation: CD39 gene transfer augments nucleoside triphosphate diphosphohydrolase activity in injured rabbit arteries. Surgery 130:296–303

    Article  PubMed  CAS  Google Scholar 

  • Gaziano JM, Gaziano TA, Glynn RJ et al (2000) Light-to-moderate alcohol consumption and mortality in the physicians’ health study enrollment cohort. J Am Col Cardiol 35:96–105

    Article  CAS  Google Scholar 

  • Goldberg DM, Hahn SE, Parkes JG (1995) Beyond alcohol: beverage consumption and cardiovascular mortality. Clin Chim Acta 237:155–187

    Article  PubMed  CAS  Google Scholar 

  • Gonzales-Calvin JL, Saunders JB, Williams R (1983) Effects of ethanol and acetaldehyde on hepatic plasma membrane ATPases. Biochem Pharmacol 32:1723–1728

    Article  Google Scholar 

  • Green RJ, Baron DN (1986) The acute in vitro effect of ethanol, its metabolites and other toxic alcohols on ion flux in isolated human leucocytes and erythrocites. Biochem Pharmacol 35:3457–3464

    Article  PubMed  CAS  Google Scholar 

  • Guerri C, Grisolia S (1983) Chronic ethanol treatment affects synaptosomal membrane-bound enzymes. Pharmacol Biochem Beh 18:45–50

    Article  CAS  Google Scholar 

  • Heymann D, Reddington M, Kreutzberg GW (1984) Subcellular localization of 5′-nucleotidase in rat brain. J Neurochem 43:263–273

    Article  Google Scholar 

  • Hillbom M, Kangasaho M, Löwbeer C et al (1985) Effects of ethanol on platelet function. Alcohol 2:429–432

    Article  PubMed  CAS  Google Scholar 

  • Kannel WB, Ellison RC (1996) Alcohol and coronary heart disease: the evidence for a protective effect. Clin Chim Acta 246:59–76

    Article  PubMed  CAS  Google Scholar 

  • Kawashima Y, Nagasawa T, Nimomiya H (2000) Contribution of ecto- 5′- nucleotidase to the inhibition platelet aggregation by human endothelial cells. Blood 96:2157–2162

    PubMed  CAS  Google Scholar 

  • Lacoste L, Hung J, Lam JT (2001) Acute and delayed antithrombotic effects of alcohol in humans. Am J Cardiol 87:82–85

    Article  PubMed  CAS  Google Scholar 

  • Landolfi R, Steiner M (1984) Ethanol raises prostacyclin in vivo and in vitro. Blood 64:679–682

    PubMed  CAS  Google Scholar 

  • Léon C, Hechler B, Vial C et al (1997) The P2Y receptor is an ADP receptor antagonized by ATP and expressed in platelets and megakaryoblastic cells. FEBS Lett 403:26–31

    Article  PubMed  Google Scholar 

  • Lunkes GI, Lunkes D, Stefanello F et al (2003) Enzymes that hydrolyze adenine nucleotides in diabetes and associated pathologies. Thromb Res 109:189–194

    Article  PubMed  CAS  Google Scholar 

  • Macieira MS, Almeida WG, Silva EA et al (1997) Alcohol dependence induced in rats by semivoluntary intermittent. Braz J Medic Biolog Res 30:1107–1111

    CAS  Google Scholar 

  • Malatová Z, Císková D (2002) Effect of ethanol on axonal transport of cholinergic enzymes in rat sciatic nerve. Alcohol 26:115–120

    Article  PubMed  Google Scholar 

  • Marcus A, Broekman MJ, Drosopoulos JHF et al (2001) Inhibition of platelet recruitment by endothelial cell CD 39/ecto-ADPase: significance for occlusive vascular diseases. Ital Hearth J 2:824–830

    CAS  Google Scholar 

  • Marques A, Guerri C (1988) Effects of ethanol on rat brain (Na+, K+) ATPase from native and delipidized synaptic membranes. Biochem Pharmacol 37:601–606

    Article  PubMed  CAS  Google Scholar 

  • Mattson MP (2008) Hormesis defined. Ageing Res Rev 7:1–7

    Article  PubMed  CAS  Google Scholar 

  • Mehta P, Mehta J, Lawson D, Patel S (1987) Ethanol stimulates prostacyclin biosynthesis by human neutrophils and potentiates anti-platelet aggregatory effects of prostacyclin. Thromb Res 48:171–180

    Article  Google Scholar 

  • Meister KA, Whelan EM, Kava R (2000) The health effects of moderate alcohol intake in humans: an epidemiologic review. Critic Rev Clin Labor Scien 37:261–296

    Article  CAS  Google Scholar 

  • Mikhailidis DP, Barradas MA, Jeremy JY (1990) The effect of ethanol on platelet function and vascular prostanoids. Alcohol 7:171–180

    Article  PubMed  CAS  Google Scholar 

  • Nguyen A, Packham MA, Rand ML (1999) Effects of ethanol on platelet responses associated with adhesion to collagen. Thromb Res 95:303–314

    Article  PubMed  CAS  Google Scholar 

  • Pereira SRC, Menezes GA, Franco GC et al (1998) Chronic ethanol consumption impairs spatial remote memory in rats but does not affect cortical cholinergic parameters. Pharmacol Biochem Behav 60:305–311

    Article  PubMed  CAS  Google Scholar 

  • Pilla C, Emanuelli T, Frasseto SS et al (1996) ATP diphosphohydrolase activity (apyrase, EC 3.6.1.5.) in human blood platelets. Platelets 7:225–230

    Article  CAS  Google Scholar 

  • Pinsky DJ, Broekman MJ, Peschon JJ et al (2002) Elucidation of the thromboregulatory role of CD 39 ecto-apyrase in the ischemic brain. J Clinic Invest 109:1031–1040

    CAS  Google Scholar 

  • Ramamurthi A, Robson SC, Lewis RS (2001) Effects of nitric oxide (NO) and soluble nucleoside triphosphate diphosphohydrolase (NTPDase) on inhibition of platelet deposition in vitro. Thromb Res 102:331–341

    Article  PubMed  CAS  Google Scholar 

  • Renaud SC, Ruf JC (1996) Effects of alcohol on platelet functions. Clin Chim Acta 246:77–89

    Article  PubMed  CAS  Google Scholar 

  • Rico EP, Rosemberg DB, Senger MR et al (2008) Ethanol and acetaldehyde alter NTPDase and 5′-nucleotidase from zebrafish brain membranes. Neurochem Int 52:290–296

    Article  PubMed  CAS  Google Scholar 

  • Robson S, Sévigny J, Zimmermann H (2006) The E-NTPDase family of ectonucleotidases: structure function relationships and pathophysiological significance. Purinergic Signal 2:409–430

    Article  PubMed  CAS  Google Scholar 

  • Rodrigo R, Thielemann L (1997) Effects of chronic and acute ethanol exposure on renal (Na+, K+) ATPase in the rat. Gen Pharmacol 29:719–723

    PubMed  CAS  Google Scholar 

  • Rubin R (1989) Ethanol interferes with collagen-induced platelet activation by inhibition of arachidonic acid mobilization. Arch Biochem Bioph 270:99–113

    Article  CAS  Google Scholar 

  • Sanchez-Amate MC, Carrasco MP, Marco C et al (1996) Adaptative changes induced by chronic ethanol ingestion on hepatic mitochondrial and microssomal enzyme activities. Intern J Biochem Cel Biol 28:23–27

    Article  CAS  Google Scholar 

  • Schetinger MR, Morsch VM, Bonan C et al (2007) NTPDase and 5′-nucleotidase activities in physiological and disease conditions: new perspectives for human health. Biofactors 31:77–98

    Article  PubMed  CAS  Google Scholar 

  • Sévigny J, Sundberg C, Braun N et al (2002) Differential catalytic properties and vascular topography of murine nucleoside triphosphate diphosphohydrolase 1 (NTPDase 1) and NTPDase 2 have implicatios for thromboregulation. Blood 99:2801–2809

    Article  PubMed  Google Scholar 

  • Soslau G, Youngprapakorn D (1997) A possible dual physiological role of extracellular ATP in the modulation of platelet aggregation. Biochim Bioph Acta 1355:131–140

    Article  CAS  Google Scholar 

  • Soslau G, McKenzie RJ, Brodsky I et al (1995) Extracellular ATP inhibits agonist-induced mobilization of internal calcium in human platelets. Biochim Bioph Acta 1268:73–80

    Article  Google Scholar 

  • Soslau G, Schechner AJ, Alcasid LPJ et al (2000) Influence of vortex speed on fresh versus stored platelet aggregation in the absence and presence of extracellular ATP. Thromb Res 97:15–27

    Article  PubMed  CAS  Google Scholar 

  • Swann A (1987) (Na+, K+)-ATPase and noradrenergic function: effects of chronic ethanol. Eur J Pharmacol 134:145–153

    Article  PubMed  CAS  Google Scholar 

  • Takahashi O (2000) Characteristics of rat platelets and relative contributions of platelets and blood coagulation to haemostasis. Food Chem Toxicol 38:203–218

    Article  PubMed  CAS  Google Scholar 

  • Yegutkin G (2008) Nucleotide and nucleoside converting ectoenzymes: important modulators of purinergic signaling cascade. Biochim Biophys Acta 1783:673–694

    Article  PubMed  CAS  Google Scholar 

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Acknowledgment

This study was supported by CNPq, FAPERGS, CAPES and the Federal University of Santa Maria, RS, Brazil.

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Correspondence to Vera Maria Morsch.

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Dias, G.R.M., Schetinger, M.R.C., Spanevello, R. et al. Hormetic acute response and chronic effect of ethanol on adenine nucleotide hydrolysis in rat platelets. Arch Toxicol 83, 263–269 (2009). https://doi.org/10.1007/s00204-008-0395-6

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  • DOI: https://doi.org/10.1007/s00204-008-0395-6

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