Abstract
It has been demonstrated that a wide variety of white blood cells and macrophages (i.e. Kupffer cells, alveolar and peritoneal macrophages and neutrophils) contain glycine-gated chloride channels. Binding of glycine on the receptor stimulates Cl− influx causing membrane hyperpolarization that prevents agonist-induced influx of calcium. Since platelet-aggregation is calcium-dependent, this study was designed to test the hypothesis that glycine would inhibit platelet aggregation. Rats were fed diets rich of glycine for 5 days, while controls received isonitrogenous valine. The bleeding time and ADP- and collagen-induced platelet aggregation were measured. Dietary glycine significantly increased bleeding time about twofold compared to valine-treated controls. Furthermore, the amplitude of platelet aggregation stimulated with ADP or collagen was significantly decreased in whole blood drawn from rats fed 2.5 or 5 % dietary glycine by over 50 %. Addition of glycine in vitro (1–10 mM) also blunted rat platelet aggregation in a dose-dependent manner. Strychnine, a glycine receptor antagonist, abrogated the inhibitory effect of glycine on platelet-aggregation in vitro suggesting the glycine works via a glycine receptor. Glycine also blunted aggregation of human platelets. Further, the glycine receptor was detected in both rat and human platelets by western blotting. Based on these data, it is concluded that glycine prevents aggregation of platelets in a dose-dependent manner via mechanisms involving a glycine receptor.
Similar content being viewed by others
References
Abrahamsen AF, Eika C, Godal HC, Lorentsen E (1974) Effect of acetylsalicylic acid and dipyridamole on platelet survival and aggregation in patients with atherosclerosis obliterans. Scand J Haematol 13:241–245
Aono J, Sakai K (1986) Effects of a new antihypertensive agent, SGB-1534, on rat platelet aggregation. Jpn J Pharmacol 42:493–500
Becker RC (1991) Seminars in thrombosis, thrombolysis and vascular biology. 3. Platelet activity in cardiovascular disease. Cardiology 79:49–63
Bertuccio P et al (2011) Coronary heart disease and cerebrovascular disease mortality in young adults: recent trends in Europe. Eur J Cardiovasc Prev Rehabil 18:627–634
Betz H, Becker CM (1988) The mammalian glycine receptor: biology and structure of a neuronal chloride channel protein. Neurochem Int 13:137–146
Blednov YA, Bleck V, Harris RA (1996) Measurement of glycine receptor function by radioactive chloride uptake. J Neurosci Methods 68:253–257
Bruschi G et al (1985) Cytoplasmic free [Ca2+] is increased in the platelets of spontaneously hypertensive rats and essential hypertensive patients. Clin Sci (Lond) 68:179–184
Choi IS et al (2009) Cyclic AMP-mediated long-term facilitation of glycinergic transmission in developing spinal dorsal horn neurons. J Neurochem 110:1695–1706
Cooper RS, Shamsi N, Katz S (1987) Intracellular calcium and sodium in hypertensive patients. Hypertension 9:224–229
Cronberg S, Caen JP (1970) Mg plus plus- and Ca plus plus-induced platelet aggregation and ADP. Thromb Diath Haemorrh 24:432–437
Curtis DR, Duggan AW (1969) The depression of spinal inhibition by morphine. Agents Actions 1:14–19
Curtis DR, Hosli L, Johnston GA (1968a) A pharmacological study of the depression of spinal neurones by glycine and related amino acids. Exp Brain Res 6:1–18
Curtis DR, Hosli L, Johnston GA, Johnston IH (1968b) The hyperpolarization of spinal motoneurons by glycine and related amino acids. Exp Brain Res 5:235–258
Danober L, Pape HC (1998) Strychnine-sensitive glycine responses in neurons of the lateral amygdala: an electrophysiological and immunocytochemical characterization. Neuroscience 85:427–441
Dejana E, Villa S, de Gaetano G (1982) Bleeding time in rats: a comparison of different experimental conditions. Thromb Haemost 48:108–111
Ferri A, Calza R, Pellegrini A, Cattani L (1994) Two distinct mechanisms of inhibition of platelets aggregation by acetylsalicylic acid. Biochem Mol Biol Int 32:1101–1107
Flores NA (1996) Platelet activation during myocardial ischaemia: a contributory arrhythmogenic mechanism. Pharmacol Ther 72:83–108
Froh M, Thurman RG, Wheeler MD (2002) Molecular evidence for a glycine-gated chloride channel in macrophages and leukocytes. Am J Physiol Gastrointest Liver Physiol 283:G856–G863
Fuster V, Badimon L, Badimon JJ, Chesebro JH (1992) The pathogenesis of coronary artery disease and the acute coronary syndromes (2). N Engl J Med 326:310–318
Gori T (2011) Viscosity, platelet activation, and hematocrit: progress in understanding their relationship with clinical and subclinical vascular disease. Clin Hemorheol Microcirc 49:37–42
Groseclose SL (2001) Mortality from coronary heart disease and acute myocardial infarction–United States, 1998. MMWR Morb Mortal Wkly Rep 50:90–93
Gulati S, Khullar M, Sharma BK, Ganguly NK (1996) Platelet calcium pump activity in essential hypertensives and their first-degree relatives. Mol Cell Biochem 156:37–42
Hondo M, Furutani N, Yamasaki M, Watanabe M, Sakurai T (2011) Orexin neurons receive glycinergic innervations. PLoS ONE 6:e25076
Ikejima K, Qu W, Stachlewitz RF, Thurman RG (1997) Kupffer cells contain a glycine-gated chloride channel. Am J Physiol 272:G1581–G1586
Jancinova V, Nosal R, Petrikova M (1992) Evidence on the role of three calcium pools in Ca-ionophore A23187-stimulated rat blood platelet aggregation. Cell Signal 4:525–530
Kaliman J, Sinzinger H, Joskowicz G (1983) Clinical value of platelet migration test in patients with peripheral vascular disease. Vasa 12:357–362
Lynch JW (2004) Molecular structure and function of the glycine receptor chloride channel. Physiol Rev 84:1051–1095
Mills DG, Philp RB, Hirst M (1974) The effects of some salicylate analogues on human blood platelets. 1. Structure activity relationships and the inhibition of platelet aggregation. Life Sci 14:659–672
O’Brien JR (1964) The mechanism and prevention of platelet adhesion and aggregation considered in relation to arterial thrombosis. Blood 24:309–314
Pfeiffer F, Simler R, Grenningloh G, Betz H (1984) Monoclonal antibodies and peptide mapping reveal structural similarities between the subunits of the glycine receptor of rat spinal cord. Proc Natl Acad Sci USA 81:7224–7227
Poch E et al (1993) Intracellular calcium mobilization and activation of the Na+/H+ exchanger in platelets. Biochem J 290(Pt 2):617–622
Poch E et al (1994) Intracellular calcium concentration and activation of the Na+/H+ exchanger in essential hypertension. Kidney Int 45:1037–1043
Qi RB et al (2007) Glycine receptors contribute to cytoprotection of glycine in myocardial cells. Chin Med J (Engl) 120:915–921
Rosse RB et al (1989) Glycine adjuvant therapy to conventional neuroleptic treatment in schizophrenia: an open-label, pilot study. Clin Neuropharmacol 12:416–424
Shechter M et al (1999) Oral magnesium supplementation inhibits platelet-dependent thrombosis in patients with coronary artery disease. Am J Cardiol 84:152–156
Siffert W (1995) Regulation of platelet function by sodium–hydrogen exchange. Cardiovasc Res 29:160–166
Stachlewitz RF et al (2000) Glycine inhibits growth of T lymphocytes by an IL-2-independent mechanism. J Immunol 164:176–182
Werman R, Davidoff RA, Aprison MH (1967) Inhibition of motoneurones by iontophoresis of glycine. Nature 214:681–683
Wheeler MD et al (1999) Glycine: a new anti-inflammatory immunonutrient. Cell Mol Life Sci 56:843–856
Wheeler MD et al (2000) Dietary glycine blunts lung inflammatory cell influx following acute endotoxin. Am J Physiol Lung Cell Mol Physiol 279:L390–L398
Young AB, Snyder SH (1973) Strychnine binding associated with glycine receptors of the central nervous system. Proc Natl Acad Sci USA 70:2832–2836
Zahavi M, Zahavi J, Kakkar VV (1984) The effect of pyridoxal-5-phosphate on the inhibition of platelet aggregation and adenosine-3′-5′-cyclic monophosphate accumulation in human platelets. Life Sci 35:1497–1503
Zhong Z, Jones S, Thurman RG (1996) Glycine minimizes reperfusion injury in a low-flow, reflow liver perfusion model in the rat. Am J Physiol 270:G332–G338
Zucker HD (1949) Platelet thrombosis in human hemostasis; a histologic study of skin wounds in normal and purpuric individuals. Blood 4:631–645
Acknowledgments
This work was supported, in part, by grant from National Institution of Health and Peter Schemmer was supported by Deutsche Forschungsgemeinschaft (SCHE 521/7-1).
Author information
Authors and Affiliations
Corresponding author
Additional information
P. Schemmer and Z. Zhong contributed equally to this work.
Ronald G. Thurman: deceased.
Rights and permissions
About this article
Cite this article
Schemmer, P., Zhong, Z., Galli, U. et al. Glycine reduces platelet aggregation. Amino Acids 44, 925–931 (2013). https://doi.org/10.1007/s00726-012-1422-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00726-012-1422-8