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Effect of methionine feeding on oxidative stress, intracellular calcium and contractility in cardiomyocytes isolated from male and female rats

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Abstract

Homocysteine (Hcy) is a breakdown product of methionine metabolism. The risk of cardiovascular disease (CVD) correlates with an increase in plasma Hcy levels. The aim of this study was to investigate whether 1% methionine supplementation of adult rats altered intracellular reactive oxygen species (ROS) generation, intracellular Ca2+ content, and contractile activity in freshly isolated cardiomyocytes. This was measured under normal conditions and during oxidative stress in freshly isolated cardiomyocytes. Single rat cardiomyocytes from both sexes were isolated by enzymatic and mechanical dispersion techniques. Fluorescence microscopy was used to measure ROS production and intracellular Ca2+ concentration. Cell contraction was measured using a video camera. During exposure to 200 μM, H2O2 female cardiomyocytes produced significantly fewer ROS and had a higher intracellular Ca2+ concentration compared to male cardiomyocytes in control and methionine-fed conditions. The contractility of cardiomyocytes isolated from male rats was insignificantly decreased after methionine feeding compared to control, while the contractility of cardiomyocytes from female rats insignificantly reduced after methionine feeding and acute exposure to oxidative stress. These findings provide evidence that during exposure to 200 μM H2O2, cardiomyocytes from female rats produce less ROS and have higher intracellular Ca2+ levels. There were no significant effects on contractility in cardiomyocytes from either gender and under any of the different conditions.

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References

  1. Dubois-Deruy E, Peugnet V, Turkieh A, Pinet F (2020) Oxidative stress in cardiovascular diseases. Antioxidants 9:864

    Article  CAS  Google Scholar 

  2. Wang R, Wang M, He S, Sun G, Sun X (2020) Targeting calcium homeostasis in myocardial ischaemia/reperfusion injury: an overview of regulatory mechanisms and therapeutic reagents. Front Pharmacol 11:872

    Article  CAS  Google Scholar 

  3. King N, McGivan JD, Griffiths EJ, Halestrap AP, Suleiman M-S (2003) Glutamate loading protects freshly isolated and perfused adult cardiomyocytes against intracellular ROS generation. J Mol Cell Cardiol 35:975–984

    Article  CAS  Google Scholar 

  4. British Heart Foundation UK factsheet, July 2020

  5. Lloyd-Jones DM, Larson MG, Beiser A, Levy D (1999) Lifetime risk of developing coronary heart disease. Lancet 353:89–92

    Article  CAS  Google Scholar 

  6. Austin R, Lentz S, Werstuck G (2004) Role of hyperhomocysteinemia in endothelial dysfunction and atherothrombotic disease. Cell Death Differ 11:S56–S64

    Article  CAS  Google Scholar 

  7. Carmel R, Jacobsen DW (2001) Homocysteine in health and disease. Cambridge University Press, New York

    Google Scholar 

  8. Selhub J (1999) Homocysteine metabolism. Ann Rev Nutr 19:217–246

    Article  CAS  Google Scholar 

  9. Wang X, Cui L, Joseph J, Jiang B, Pimental D, Handy DE, Liao R, Loscalzo J (2012) Homocysteine induces cardiomyocyte dysfunction and apoptosis through p38 MAPK-mediated increase in oxidant stress. J Mol Cell Cardiol 52:753–760

    Article  CAS  Google Scholar 

  10. Almashhadany A, Shackebaei D, Van der Touw T, Jones G, Suleiman M-S, King N (2015) Homocysteine exposure impairs myocardial resistance to ischaemia reperfusion and oxidative stress. Cell Physiol Biochem 37:2265–2274

    Article  CAS  Google Scholar 

  11. Fukada S-I, Shimada Y, Morita T, Sugiyama K (2006) Suppression of methionine-induced hyperhomocysteinemia by glycine and serine in rats. Biosci Biotechnol Biochem 70:2403–2409

    Article  CAS  Google Scholar 

  12. Norsidah K-Z, Asmadi AY, Azizi A, Faizah O, Kamisah Y (2013) Palm tocotrienol-rich fraction reduced plasma homocysteine and heart oxidative stress in rats fed with a high-methionine diet. J Physiol Biochem 69:441–449

    Article  CAS  Google Scholar 

  13. Demerchi SA, McFarlane JR, Moens PDJ, King N (2020) Effect of L-methionine feeding on serum homocysteine and glutathione levels in male and female Wistar rats. Adv Biochem 8:21–25

    Article  CAS  Google Scholar 

  14. King N, Korolchuk S, McGivan J, Suleiman M-S (2004) A new method of quantifying glutathione levels in freshly isolated single superfused rat cardiomyocytes. J Pharmacol Toxicol Methods 50(3):215–222

    Article  CAS  Google Scholar 

  15. King N, Lin H, McGivan JD, Suleiman M-S (2004) Aspartate transporter expression and activity in hypertrophic rat heart and ischaemia–reperfusion injury. J Physiol 556:849–858

    Article  CAS  Google Scholar 

  16. Kurebayashi N, Harkins A, Baylor S (1993) Use of fura red as an intracellular calcium indicator in frog skeletal muscle fibers. Biophys J 64:1934–1960

    Article  CAS  Google Scholar 

  17. Grill HP, Zweier JL, Kuppusamy P, Weisfeldt ML, Flaherty JT (1992) Direct measurement of myocardial free radical generation in an in vivo model: effects of postischemic reperfusion and treatment with human recombinant superoxide dismutase. J Am Coll Cardiol 20:1604–1611

    Article  CAS  Google Scholar 

  18. Zweier JL, Flaherty JT, Weisfeldt ML (1987a) Direct measurement of free radical generation following reperfusion of ischemic myocardium. Proc Natl Acad Sci 84:1404–1407

    Article  CAS  Google Scholar 

  19. Zweier JL, Rayburn BK, Flaherty JT, Weisfeldt ML (1987b) Recombinant superoxide dismutase reduces oxygen free radical concentrations in reperfused myocardium. J Clin Invest 80:1728

    Article  CAS  Google Scholar 

  20. Ihaka R, Gentleman R (1996) R: a language for data analysis and graphics. J Comput Graph Stat 5:299–314

    Google Scholar 

  21. Camper-Kirby D, Welch S, Walker A, Shiraishi I, Setchell KD, Schaefer E, Kajstura J, Anversa P, Sussman MA (2001) Myocardial Akt activation and gender increased nuclear activity in females versus males. Circ Res 88:1020–1027

    Article  CAS  Google Scholar 

  22. Guerra S, Leri A, Wang X, Finato N, Di Loreto C, Beltrami CA, Kajstura J, Anversa P (1999) Myocyte death in the failing human heart is gender dependent. Circ Res 85:856–866

    Article  CAS  Google Scholar 

  23. Mallat Z, Fornes P, Costagliola R, Esposito B, Belmin J, Lecomte D, Tedgui A (2001) Age and gender effects on cardiomyocyte apoptosis in the normal human heart. J Gerontol A: Biol Sci Med Sci 56:M719–M723

    Article  CAS  Google Scholar 

  24. Setnik B, de Souza FG, d'Almeida V, Nobrega JN (2004) Increased homocysteine levels associated with sex and stress in the learned helplessness model of depression. Pharmacol Biochem Behav 77:155–161

    Article  CAS  Google Scholar 

  25. Curl C, Wendt I, Kotsanas G (2001) Effects of gender on intracellular [Ca2+] in rat cardiac myocytes. Pflugers Arch 441:709–716

    Article  CAS  Google Scholar 

  26. Gen W, Tani M, Takeshita J, Ebihara Y, Tamaki K (2001) Mechanisms of Ca2+ overload induced by extracellular H2O2 in quiescent isolated rat cardiomyocytes. Basic Res Cardiol 96:623–629

    Article  CAS  Google Scholar 

  27. Zivkovic V, Jakovljevic V, Djordjevic D, Vuletic M, Barudzic N, Djuric D (2012) The effects of homocysteine-related compounds on cardiac contractility, coronary flow, and oxidative stress markers in isolated rat heart. Mol Cell Biochem 370:59–67

    Article  CAS  Google Scholar 

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Acknowledgements

We would like to thank the technical staff for their excellent assistance.

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This work was supported by Iraqi Ministry of Higher Education and University of New England.

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Correspondence to Nicola King.

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Demerchi, S.A., King, N., McFarlane, J.R. et al. Effect of methionine feeding on oxidative stress, intracellular calcium and contractility in cardiomyocytes isolated from male and female rats. Mol Cell Biochem 476, 2039–2045 (2021). https://doi.org/10.1007/s11010-020-04011-2

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  • DOI: https://doi.org/10.1007/s11010-020-04011-2

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