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Suppression of inositol phosphate release by cardiac myocytes isolated from fish oil-fed pigs

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Abstract

Fatty acid composition of cardiac myocytes and release of inositol phosphates in pigs fed a fish oil supplemented diet was examined. Two groups of female pigs were fed diets supplemented with either 50 g/kg diet beef tallow (as control) or 50 g/kg diet fish oil (MaxEPA) rich in n-3 fatty acids. After 6 weeks of supplementation, the pigs were anesthetized and hearts were removed. Cardiac myocytes were isolated, lipid extracted and separated into non-polar and polar lipids by thin-layer chromatography. Fatty acid composition of individual neutral and polar lipid classes were examined by gas chromatography. To study the effect of membrane phospholipid modification on the phospholipase C (PLC) mediated release of inositol phosphates, cardiac myocytes were labelled with 4 μCi/mL myo-[2-3H]inositol for 48 h. After stimulation with epinephrine and phenylephrine, the water soluble [3H]inositol products were extracted, separated from [3H]inositol and [3H]glycerophosphoinositol by chromatography on Dowex AG 1-X8 and quantitated by scintillation counting. Cardiac myocytes isolated from fish oil-fed pigs had higher levels of n-3 polyunsaturated fatty acid in the non-esterified fatty acid and phospholipid fraction. Similarly, these cardiac myocytes had increased level of n-3 fatty and decreased n-6 fatty acids in all the phospholipid fractions, PE, PC, PI and PS (p < 0.05). After stimulation, the levels of [3H]inositol trisphosphate (IP3) and [3H]inositol tetrakisphosphate (IP4) in cardiac myocytes isolated from fish oil-fed pigs were significantly reduced (p < 0.05) compared to myocytes isolated from beef tallow fed-pigs. This study for the first time has utilised adult cardiac myocytes to demonstrate the effect of n-3 PUFA supplementation on cardiac myocyte phospholipid fatty acid composition and release of second messengers.

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References

  1. McLennan PL, Abeywardena MY, Charnock J: Dietary fish oil prevents ventricular fibrillation following coronary artery occlusion and reperfusion. Am Heart J 116: 709–717, 1988

    Google Scholar 

  2. McLennan PL: Relative effects of dietary saturated, monounsaturated and polyunsaturated fatty acids on cardiac arrhythmias in rats. Am J Clin Nutr 57: 207–212, 1993

    Google Scholar 

  3. Billman GE, Hallaq H, Leaf A: Prevention of ischemia-induced ventricular fibrillation by omega 3 fatty acids. Proc Natl Acad Sci USA 91: 4427–4430, 1994

    Google Scholar 

  4. Billman GE, Kang J X, Leaf A: Prevention of ischemia induced cardiac sudden death by n-3 polyunsaturated fatty acids in dogs. Lipids 32: 1161–1168, 1997

    Google Scholar 

  5. Kang JX, Leaf A: Effects of long chain polyunsaturated fatty acids on the contraction of neonatal rat cardiac myocytes. Proc Natl Acad Sci USA 91: 9886–9890, 1994

    Google Scholar 

  6. Kang JX, Leaf A: Prevention and termination of β-adrenergic agonistinduced arrhythmias by free polyunsaturated fatty acids in neonatal rat cardiac myocytes. Biochem Biophy Res Commun 208: 629–636, 1995

    Google Scholar 

  7. Hallaq H, Smith TW, Leaf A: Modulation of dihydropyridine-sensitive calcium channels in heart cells by fish oil fatty acids. Proc Natl Acad Sci USA 89: 1760–1764, 1992

    Google Scholar 

  8. Kinsella JE, Lokesh B, Stone RA: Dietary n-3 polyunsaturated fatty acids and amelioration of cardiovascular disease: Possible mechanisms. Am J Clin Nutr 52: 1–28, 1990

    Google Scholar 

  9. Charnock JS: Lipids and cardiac arrhythmias. Prog Lipid Res 33: 355–385, 1994

    Google Scholar 

  10. Nair SSD, Leitch JW, Falconer J, Garg ML: Prevention of cardiac arrhythmia by dietary n-3 polyunsaturated fatty acids and their mechanism of action. J Nutr 127: 383–393, 1997

    Google Scholar 

  11. Nair SSD, Leitch JW, Garg ML: Cardiac n-3 non-esterified fatty acids are selectively increased in fish oil-fed pigs following myocardial ischemia. J Nutr 129: 1518–1523, 1999

    Google Scholar 

  12. Choy PC, Tran K, Hatch GM, Kroeger EA: Phospholipid metabolism in the mammalian heart. Prog Lipid Res 36: 85–101, 1997

    Google Scholar 

  13. Berridge MJ, Irvine RF: Inositol phosphates and cell signalling. Nature 341: 197–205, 1989

    Google Scholar 

  14. Nishizuka Y: Studies and perspectives of protein kinase C. Science 29: 305–312, 1986

    Google Scholar 

  15. Williams S, Meij JTA, Panagia V: Membrane phospholipids and adrenergic receptor function. Mol Cell Biochem 149/150: 217–222, 1995

    Google Scholar 

  16. Manning AS, Hearse DJ: Reperfusion induced arrhythmias: Mechanisms and prevention. J Mol Cell Cardiol 16: 497–518, 1988

    Google Scholar 

  17. de Chaffoy de Courcelles D: Is there evidence of a role of the phosphoinositol-cycle in the myocardium? Mol Cell Biochem 88: 65–72, 1989

    Google Scholar 

  18. Chapman MJ: Animal lipoproteins: Chemistry, structure and comparative aspects. J Lipid Res 231: 789–853, 1980

    Google Scholar 

  19. Harris SW: n-3 fatty acids and serum lipoproteins: Animal studies. Am J Clin Nutr 65(suppl): 1611S–1616S, 1997

    Google Scholar 

  20. Spinale FG, Mukherjee R, Fulbright MB, Hu J, Crawford FA, Zile MR: Contractile properties of isolated porcine ventricular myocytes. Cardiovasc Res 27: 304–311, 1993

    Google Scholar 

  21. Nair SSD, Leitch J, Garg ML: Specific modifications of phosphatidylinositol and non-esterified fatty acid fractions in cultured porcine cardiomyocytes supplemented with n-3 polyunsaturated fatty acids. Lipids 34: 697–704, 1999

    Google Scholar 

  22. Lepage G, Roy GC: Direct transesterification of all classes of lipid in a one step reaction. J Lipid Res 27: 114–120, 1986

    Google Scholar 

  23. Folch J, Lees M, Sloane-Stanley GHS: A simple method for the isolation and purification of total lipids from animal tissues. J Biol Chem 226: 497–509, 1957

    Google Scholar 

  24. Christie WW: Lipid Analysis. Pergamon Press, Oxford, 1982

    Google Scholar 

  25. Metcalfe LD, Schmitz AA: The rapid preparation of fatty acid esters for gas chromatographic analysis. Anal Chem 33: 363–364, 1961

    Google Scholar 

  26. Touchstone JC, Chen JC, Beaver KM: Improved separation of phospholipids in thin layer chromatography. Lipids 15: 61–62, 1980

    Google Scholar 

  27. Garg ML, Blake RJ: Cholesterol dynamics in rats fed diets containing either canola oil or sunflower oil. Nutr Res 17: 485–492, 1997

    Google Scholar 

  28. Bird IM: Analysis of cellular phosphoinositides and phosphoinositols by extraction and simple analytical procedures. Meth Mol Biol 27: 227–248, 1994

    Google Scholar 

  29. Morisaki N, Kauzaki T, Fujiyama Y, Osawa I, Shirai K, Matsuoko N, Saito Y, Yoshida S: Metabolism of n-3 polyunsaturated fatty acids and modification of phospholipids in cultured rabbit aortic smooth muscle cells. J Lipid Res 26: 930–939, 1985

    Google Scholar 

  30. Lamers JMJ, Dekkers DHW, De Jonge N, Meij JTA: Modification of fatty acid composition of the phospholipids of cultured rat ventricular myocytes and the rate of phosphatidylinositol 4,5-bisphosphate hydrolysis. J Mol Cell Cardiol 24: 605–618, 1992

    Google Scholar 

  31. Corr PB, Heathers GP, Yamada KA: Mechanisms contributing to the arrhythmogenic influences of alpha 1-adrenergic stimulation in the ischemic heart. Am J Med 87: 19S–25S, 1989

    Google Scholar 

  32. Woodcock EA, Matkovitch ST, Binah O: Ins(1,4,5)P3 and cardiac dysfunction. Cardiovasc Res 40: 251–256, 1998

    Google Scholar 

  33. Woodcock EA, Anderson KE, Du XJ, Dart AM: Effects of dietary fat supplementation on inositol phosphate release and metabolism in rat left atria. J Mol Cell Cardiol 27: 867–871, 1995

    Google Scholar 

  34. Karmazyn M, Horackova M, Murphy MG: Effects of dietary cod liver oil on fatty acid composition and calcium transport in isolated adult rat ventricular myocytes and on the response of isolated hearts to ischemia and reperfusion. Can J Physiol Pharmacol 65: 201–209, 1987

    Google Scholar 

  35. Meij JTA, Lamers JMJ: Alpha-1-adrenergic stimulation of phosphoinositide breakdown in cultured neonatal rat ventricular myocytes. Mol Cell Biochem 88: 73–75, 1989

    Google Scholar 

  36. Steinfath M, Chen YY, Lavicky J, Magnussen O, Nose M, Rosswag S, Schmitz W, Scholz H: Cardiac ?1-adrenoceptor densities in different mammalian species. Br J Pharmacol 107: 185–188, 1992

    Google Scholar 

  37. Terzic A, Puceat M, Vassort G, Vogel SM: Cardiac ?1-adrenoreceptors: An overview. Pharmacol Rev 45: 147–175, 1993

    Google Scholar 

  38. Heathers GP, Yamada KA, Pogwidz SM, Corr PB: The contribution of α-and β-adrenergic mechanisms in the genesis of arrhythmias during myocardial ischemia and reperfusion. In: H.E. Kulbertus, G. Franck (eds). Neurocardiology. Futura Publishing Co. Inc., NY, 1988

    Google Scholar 

  39. Bird GS, Putney JW: Effect of inositol 1,3,4,5-tetrakisphosphate on inositol trisphosphate-activated Ca2+ signaling in mouse lacrimal acinar cells. J Biol Chem 271: 6766–6770, 1996

    Google Scholar 

  40. Ponsard B, Durot I, Fournier A, Oudot F, Athias A, Grynberg A: Long chain polyunsaturated fatty acids influence both α-and β-adrenergic receptors. J Am Oil Chem Soc 75: 247–254, 1998

    Google Scholar 

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Authors and Affiliations

  1. Discipline of Nutrition and Dietetics, Faculty of Medicine and Health Sciences, University of Newcastle, Callaghan, NSW, 2308, Australia

    Sudheera S.D. Nair

  2. Department of Cardiology, John Hunter Hospital, New Lambton, NSW, 2299, Australia

    James Leitch

  3. Discipline of Nutrition and Dietetics, Faculty of Medicine and Health Sciences, University of Newcastle, Callaghan, NSW, 2308, Australia

    Manohar L. Garg

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  1. Sudheera S.D. Nair
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  2. James Leitch
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  3. Manohar L. Garg
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Nair, S.S., Leitch, J. & Garg, M.L. Suppression of inositol phosphate release by cardiac myocytes isolated from fish oil-fed pigs. Mol Cell Biochem 215, 57–64 (2000). https://doi.org/10.1023/A:1026538932590

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