Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Paper
  • Published:

Identifying pathways involved in leptin-dependent aggregation of human platelets

International Journal of Obesity volume 28, pages 979–984 (2004)Cite this article

Abstract

OBJECTIVE: To investigate the role of phospholipase C (PLC), phospholipase A2 (PLA2), calcium, and protein kinase C (PKC) in mediating leptin-enhanced aggregation of human platelets.

DESIGN: In vitro, ex vivo study.

SETTING: Outpatient's Service for Prevention and Treatment of Obesity at the University Hospital of Messina, Italy.

SUBJECTS: In total, 14 healthy normal-weight male (age 31.4±1.9 y; body mass index 22.7±0.6 kg/m2) subjects.

MEASUREMENTS: Adenosine diphosphate-(ADP-) induced platelet aggregation and platelet free calcium were measured after incubation of platelets with leptin alone (5–500 ng/ml), or leptin (50 and 100 ng/ml) in combination with anti-human leptin receptor long form antibody (anti-ObRb-Ab, 1:800–1:100 dilutions), PLC inhibitor U73122 (3.125–25 μM), PLA2 inhibitor AACOCF3 (1.25–10 μM), or PKC inhibitor Ro31-8220 (1.25–10 μM).

RESULTS: Platelet stimulation with leptin leads to a significant and dose-dependent increase in ADP-induced platelet aggregation and platelet free calcium concentrations. Leptin effects on both platelet aggregation and calcium mobilization were completely abated by the co-incubation with leptin and anti-ObRb-Ab. Leptin-induced platelet aggregation was dose-dependently inhibited by U73122, AACOCF3, or Ro31-8220. The effect of leptin on intracellular calcium was inhibited in a dose-dependent manner by incubation with U73122 and AACOCF3, but not with Ro31-8220.

CONCLUSIONS: Our study confirms that leptin is able to enhance ADP-induced aggregation of human platelets, and raise the possibility that PLC, PKC, PLA2, and calcium could play a relevant role in mediating the proaggregating action of leptin.

Access through your institution
Buy or subscribe

This is a preview of subscription content, access via your institution

Access options

Access through your institution

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2

Similar content being viewed by others

References

  1. Zhang Y, Proenca R, Maffei M, Barone M, Leopold L, Friedman JM . Positional cloning of the mouse ob gene and its human homologue. Nature 1994; 372: 425–432.

    Article  CAS  Google Scholar 

  2. Campfield LA, Smith FJ, Guisez Y, Devos R, Burn P . Recombinant mouse ob protein: evidence for a peripheral signal linking adiposity and central neural networks. Science 1995; 269: 546–548.

    Article  CAS  Google Scholar 

  3. Lord GM, Matarese G, Howard JK, Baker RJ, Bloom SR, Lechler RI . Leptin modulates the T-cell immune response and reverses starvation-induced immunosuppression. Nature 1998; 394: 897–901.

    Article  CAS  Google Scholar 

  4. Sierra-Honigmann MR, Nath AK, Murakami C, Garcia-Cardena G, Papapetropoulos A, Sessa WC, Madge LA, Schechner JS, Schwabb MB, Polverini PJ, Flores-Riveros JR . Biological activation of leptin as an angiogenic factor. Science 1998; 281: 1683–1686.

    Article  CAS  Google Scholar 

  5. Ducy P, Amling M, Takeda S, Priemel M, Schilling AF, Beil FT, Shen J, Vinson C, Rueger JM, Karsenty G . Leptin inhibits bone formation through a hypothalamic relay: a central control of bone mass. Cell 2000; 100: 197–207.

    Article  CAS  Google Scholar 

  6. Chehab FF, Lim ME, Lu R . Correction of the sterility defect in homozigous obese female mice by treatment with the human recombinant leptin. Nat Genet 1996; 12: 318–320.

    Article  CAS  Google Scholar 

  7. Lee GH, Proenca R, Montez JM, Carroll KM, Darvishzadeh JG, Lee JI, Friedman JM . Abnormal splicing of the leptin receptor in diabetic mice. Nature 1996; 379: 632–635.

    Article  CAS  Google Scholar 

  8. Tartaglia LA, Dembski M, Weng X, Deng N, Culpepper J, Devos R, Richards GJ, Campfield LA, Clark FT, Deeds J . Identification and expression cloning of a leptin receptor, OB-R. Cell 1995; 83: 1263–1271.

    Article  CAS  Google Scholar 

  9. Haynes WG, Morgan DA, Walsh SA, Sivitz WI, Mark AL . Cardiovascular consequences of obesity: role of leptin. Clin Exp Pharmacol Physiol 1998; 25: 65–69.

    Article  CAS  Google Scholar 

  10. Wallace AM, McMahon AD, Packard CJ, Kelly A, Shepherd J, Gaw A, Sattar N . Plasma leptin and the risk of cardiovascular disease in the west of Scotland coronary prevention study (WOSCOPS). Circulation 2001; 104: 3052–3056.

    Article  CAS  Google Scholar 

  11. Konstantinides S, Schafer K, Koschnick S, Loskutoff DJ . Leptin-dependent platelet aggregation and arterial thrombosis suggests a mechanism for atherothrombotic disease in obesity. J Clin Invest 2001; 108: 1533–1540.

    Article  CAS  Google Scholar 

  12. Bodary PF, Westrick RJ, Wickenheiser KJ, Shen Y, Eitzman DT . Effect of leptin on arterial thrombosis following vascular injury in mice. JAMA 2002; 287: 1706–1709.

    Article  CAS  Google Scholar 

  13. Nakata M, Yada T, Soejima N, Maruyama I . Leptin promotes aggregation of human platelets via the long form of its receptor. Diabetes 1999; 48: 426–429.

    Article  CAS  Google Scholar 

  14. Corsonello A, Malara A, Ientile R, Corica F . Leptin enhances adenosine diphosphate-induced platelet aggregation in healthy subjects. Obes Res 2002; 10: 306.

    Article  Google Scholar 

  15. Puri RN . Phospholipase A2: its role in ADP- and thrombin-induced platelet activation mechanisms. Int J Biochem Cell Biol 1998; 30: 1107–1122.

    Article  CAS  Google Scholar 

  16. Corica F, Corsonello A, Perticone F . Effects of leptin on platelet function in obese patients. JAMA 2002; 288: 313–314.

    Article  Google Scholar 

  17. Corsonello A, Perticone F, Malara A, De Domenico D, Loddo S, Buemi M, Ientile R, Corica F . Leptin-dependent platelet aggregation in healthy, overweight and obese subjects. Int J Obes Relat Metab Disord 2003; 27: 566–573.

    Article  CAS  Google Scholar 

  18. Trenchard PM, Morgan RA . Sub-ambient aggregometry: a tool for examining platelet storage at any temperature between 4–22°C. Haemostasis 1986; 16: 352–361.

    CAS  PubMed  Google Scholar 

  19. Lockhart LK, McNicol A . The phospholipase C inhibitor U73122 inhibits phorbol ester-induced platelet activation. J Pharmacol Exp Ther 1999; 289: 721–728.

    CAS  PubMed  Google Scholar 

  20. Lockhart LK, Pampolina C, Nickolaychuk BR, McNicol A . Evidence for a role for phospholipase C, but not phospholipase A2, in platelet activation in response to low concentrations of collagen. Thromb Haemost 2001; 85: 882–889.

    Article  CAS  Google Scholar 

  21. Trovati M, Anfossi G . Insulin, insulin resistance and platelet function: similarities with insulin effects on cultured vascular smooth cells. Diabetologia 1998; 41: 609–622.

    Article  CAS  Google Scholar 

  22. Takekoshi K, Ishii K, Kawakami Y, Isobe K, Nanmoku T, Nakai T . Ca2+ mobilization, tyrosine hydoxylase activity, and signaling mechanisms in cultured porcine adrenal medullary chromaffin cells: effects of leptin. Endocrinology 2001; 142: 290–298.

    Article  CAS  Google Scholar 

  23. Siess W . Molecular mechanisms of platelet activation. Physiol Rev 1989; 69: 58–178.

    Article  CAS  Google Scholar 

  24. Brass LF, Manning DR, Cichowski K, Abrahams CS . Signaling through G proteins in platelets: to the integrins and beyond. Thromb Haemostasis 1997; 78: 581–589.

    Article  CAS  Google Scholar 

  25. Banno Y, Nakashima S, Ozhawa M, Nozawa Y . Differential translocation of phospholipase C isozymes to integrin-medicated cytoskeletal complexes in thrombin-stimulated human platelets. J Biol Chem 1996; 271: 14989–14994.

    Article  CAS  Google Scholar 

  26. Nishizuka Y . The role of protein kinase C in cell surface signal transdiction and tumor promotion. Nature (London) 1984; 308: 693–698.

    Article  CAS  Google Scholar 

  27. Berridge MJ . Inositol triphosphate and calcium signaling. Nature (London) 1993; 361: 315–325.

    Article  CAS  Google Scholar 

  28. Kaibuchi K, Takay Y, Swamura M, Hoshijima M, Fujikura T, Nishizuka Y . Synergistic functions of protein phosphorylation and calcium mobilization in platelet activation. J Biol Chem 1983; 258: 6701–6704.

    CAS  Google Scholar 

  29. Brune B, Ullrich V . Different calcium pools in human platelets and their role in thromboxane A2 formation. J Biol Chem 1991; 266: 19232–19237.

    CAS  Google Scholar 

  30. Brune B, von Appen F, Ullrich V . Receptor occupancy regulates Ca2+ entry and intracellular Ca2+ redistribution in activated human platelets. Biochem J 1994; 304: 993–999.

    Article  CAS  Google Scholar 

  31. Reilly M, Fitzgerald GA . Cellular activation by thromboxane A2 and other eicosanoids. Eur Heart J 1993; 14 (Suppl K): 88–93.

    CAS  PubMed  Google Scholar 

  32. Li Z, Xi X, Du X . A mitogen activated protein kinase dependent signaling pathways in the activation of platelets integrine a IIIb B3. J Biol Chem 2001; 276: 42226–42232.

    Article  CAS  Google Scholar 

  33. Quinton TM, Kim S, Dangelmaier C, Dorsam RT, Jin J, Daniel JL, Kunapuli SP . Protein Kinase C- and calcium-regulated pathways independently sinergize with Gi pathways in agonist induced fibrinogen receptor activation. Biochem J 2002; 368: 535–543.

    Article  CAS  Google Scholar 

  34. Bhalla US, Ram PI, Iyenger R . MAP Kinase phosphates as a locus of flexibility in a mitogen activated protein kinase signalling network. Science 2002; 297: 1018–1023.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work is partially supported by grants from the Italian Ministry for Education, University and Scientific Research (MIUR).

This paper is dedicated to the memory of Francesco Corsonello, MD, who spent all of his time in research on Geriatric Medicine at the Italian National Research Center on Aging (INRCA).

Author information

Authors and Affiliations

  1. Italian National Research Centres on Aging (INRCA), Cosenza, I-87100, Italy

    A Corsonello

  2. Department of Internal Medicine, University of Messina, Messina, Italy

    A Malara, D De Domenico, M Buemi & F Corica

  3. Department of Clinical and Experimental Medicine, University Magna Graecia, Catanzaro, Italy

    F Perticone

  4. Department of Experimental Pathology and Microbiology, University of Messina, Messina, Italy

    A Valenti

  5. Department of Biochemical, Physiological and Nutritional Science, University of Messina, Messina, Italy

    R Ientile

Authors
  1. A Corsonello
    View author publications

    Search author on:PubMed Google Scholar

  2. A Malara
    View author publications

    Search author on:PubMed Google Scholar

  3. D De Domenico
    View author publications

    Search author on:PubMed Google Scholar

  4. F Perticone
    View author publications

    Search author on:PubMed Google Scholar

  5. A Valenti
    View author publications

    Search author on:PubMed Google Scholar

  6. M Buemi
    View author publications

    Search author on:PubMed Google Scholar

  7. R Ientile
    View author publications

    Search author on:PubMed Google Scholar

  8. F Corica
    View author publications

    Search author on:PubMed Google Scholar

Corresponding author

Correspondence to A Corsonello.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Corsonello, A., Malara, A., De Domenico, D. et al. Identifying pathways involved in leptin-dependent aggregation of human platelets. Int J Obes 28, 979–984 (2004). https://doi.org/10.1038/sj.ijo.0802722

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue date:

  • DOI: https://doi.org/10.1038/sj.ijo.0802722

Keywords

This article is cited by

Search

Advanced search

Quick links