Skip to main content
SpringerLink
Log in

Inhibition of collagen, and thrombin-induced platelet aggregation by Lansberg’s hognose pit viper (Porthidium lansbergii hutmanni) venom

  • Published:
Journal of Thrombosis and Thrombolysis Aims and scope Submit manuscript

Abstract

The Porthidium genus is represented by the P. lansbergii rozei and P. lansbergii hutmanni (Plh) subspecies in Venezuela. The venom components of these have been little studied, probably due to the low incidence of reported accidents, although acute and serious local effects such as invasive edema and disseminated ecchymosis are present during human envenonation. The aim of this work was to characterize the in vitro effects of crude P. l. hutmanni venom, and its fractions, on platelet aggregation triggered by two physiologic agonists: thrombin and collagen. The effects of thrombin and collagen were observed on a platelet-rich plasma (PRP) solution (3 × 105 platelets/μL) using serial dilutions of P. l. hutmanni venom (0.625–40 μg). The crude venom was fractionated by anionic exchange chromatography and two peaks obtained. Crude venom and both fractions were highly inhibitory on platelet aggregation mediated by the two agonists. The anti-aggregating dose (AD50) for both agonists was determined. PRP collagen-triggered aggregation was most inhibited by the crude venom (AD50 = 0.67 μg) when compared with PRP thrombin-triggered aggregation (AD50 = 4.92 μg). Collagen-induced aggregation was more intensely inhibited by venom than thrombin-induced aggregation. In conclusion, to specify the inhibition mechanisms involved for each of the active components in the venom from these subspecies, we must characterize and purify the inhibitors of aggregation from P. l. hutmanni venom, with the purpose of suggesting new pharmacological substances to be incorporated into the therapeutic arsenal to treat hemostatic pathologies related to high levels of platelet aggregation.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Boquet P (1992) History of snake venom research. Handb Exp Pharmacol 52:3–14

    Google Scholar 

  2. Andrews R, Berndt M (2000) Snake venom modulators of platelet adhesion receptors and their ligands. Toxicon 38:775–791

    Article  PubMed  CAS  Google Scholar 

  3. Bryan J, Dangelmaier C, Selak M (1991) Identification of 50 kDa snake venom protein which specifically inhibit platelet aggregation on collagen. FEBS 283:307–310

    Article  Google Scholar 

  4. Fujimura Y, Ikeda Y, Miura S et al (1995) Isolation and characterization of jararaca GPIb-BP, a snake venom antagonist specific to platelet glycoprotein Ib. Thromb Haemost 74:743–750

    PubMed  CAS  Google Scholar 

  5. Fuly A, Machado O, Alves E, Carlini C (1997) Mechanism of inhibitory action on platelet activation of a phospholipase A2 isolated from Lachesis muta (Bushmaster) snake venom. Thromb Haemost 78:1372–1380

    PubMed  CAS  Google Scholar 

  6. Huang T, Wu Y, Ouyang C (1987) Characterization of a potent platelet aggregation inhibitor from Agkistrodon rhodostoma snake venom. Biochim Biophys Acta 925:248–257

    PubMed  CAS  Google Scholar 

  7. Fuentes O, Rodríguez-Acosta A (1997) Sobre los generos Bothriechis, Bothriopsis, Bothrops y Porthidium (Serpentes: Crotalidae) existentes en Venezuela. Claves para su identificacion de interes biomedico. Acta Biol Ven 17:31–38

    Google Scholar 

  8. Otero R, Osorio R, Valderrama R, Giraldo C (1992) Efectos Farmacológicos y Enzimáticos de los Venenos de las Serpientes de Antioquia y Choco (Colombia). Toxicon 30:611–620

    Article  PubMed  CAS  Google Scholar 

  9. Otero R, Gutiérrez J, Rojas G et al (1999) A randomized blinded clinical trial of two antivenoms, prepared by caprilic acid or ammonium sulphate fraction of IgG, in Bothrops and Porthidium snake bites in Colombia: correlation between safety and biochemical characteristics of antivenoms. Toxicon 37:895–908

    Article  PubMed  CAS  Google Scholar 

  10. Otero R, Fonnegra R, Jiménez S et al (2000) Snakebites and ethnobotany in the northwest region of Colombia. J Ethnopharmacol 71:493–504

    Article  PubMed  CAS  Google Scholar 

  11. Laemmli UK (1970) Cleavage of structural protein during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  PubMed  CAS  Google Scholar 

  12. Lowry OH, Rosebrough NJ, Farr AL, Randal JH (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275

    PubMed  CAS  Google Scholar 

  13. Kini M, Evans H (1990) Effects of snake venom proteins on blood platelet. Toxicon 28:1387–1422

    Article  PubMed  CAS  Google Scholar 

  14. Kemparaju K, Krishnakanth T, Gowda T (1999) Purification and characterization of a platelet aggregation inhibitor acidic phospholipase A2 from Indian saw-scaled viper (Echis carinatus). Toxicon 37:1659–1671

    Article  PubMed  CAS  Google Scholar 

  15. Huang T, Chiang H (1994) Effect on human platelet aggregation of phospholipase A2 purified from Heloderma horridum (beaded lizard) venom. Biochim Biophys Acta 1211:61–68

    PubMed  CAS  Google Scholar 

  16. Kamiguti A, Laing G, Lowe G, Zuzel M, Warrell D, Theakston R (1994) Biological properties of the venom of the Papuan black snake (Pseudechis papuanus): presence of a phospholipase A2 platelet inhibitor. Toxicon 32:915–925

    Article  PubMed  CAS  Google Scholar 

  17. Satish S, Tejaswini J, Krishnakantha TP, Gowda TV (2004) Purification of a class B1 platelet aggregation inhibitor phospholipase A2 from Indian cobra (Naja naja). Biochimie 86:203–210

    Article  PubMed  CAS  Google Scholar 

  18. Colman RB, Hirsh J, Mader VJ, Salzman EW (1982) Hemostasis and thrombosis. Philadelphia, PA: Lippincott

  19. Ouyang C, Huang T (1983) Potent platelet aggregation inhibitor from Trimeresurus gramineus snake venom. Biochem Biophys Acta 757:332–341

    PubMed  CAS  Google Scholar 

  20. Watson S (1999) Collagen receptor signaling in platelets and megakaryocytes. Thromb Haemost 82:365–376

    PubMed  CAS  Google Scholar 

  21. Roberts H, Monroe D III, Hoffman M (2001) Williams, hematology, 6th edn. McGraw-Hill, Inc, USA

    Google Scholar 

  22. Parise L, Smyth S, Coller B (2001) Williams, hematology, 6th edn. McGraw-Hill, Inc, USA

    Google Scholar 

  23. Navarrete LF, Vargas AM, Scannone H, Rodríguez-Acosta A (2004) Actividades hemorrágicas y necrosantes de los venenos de Porthidium l. hutmanni y P. l. rozei de dos regiones de Venezuela. IX Jornades Herpetologiques Catalanes. Soc Catalana D¨Herpetol. Barcelona, España

  24. Uma B, Veerabasappa G (2000) Molecular mechanism of lung hemorrhage induction by VRV-PL-VIIIa from Russel’s viper (Vipera russelli) venom. Toxicon 38:1129–1147

    Article  PubMed  CAS  Google Scholar 

  25. Ouyang C, Huang T (1986) Platelet aggregation inhibitors from Agkistrodon acutus snake venom. Toxicon 34:1099–1106

    Article  Google Scholar 

  26. Smith JB, Dangelmaier C, Selak M (1991) Identification of 50 kDa snake venom protein which specifically inhibit platelet aggregation on collagen. FEBS 283:307–310

    Article  CAS  Google Scholar 

  27. Soto JG, White SA, Reyes SR, Regalado R, Sanchez EE, Perez JC (2007) Molecular evolution of PIII-SVMP and RGD disintegrin genes from the genus Crotalus. Gene 389:66–72

    Article  PubMed  CAS  Google Scholar 

  28. Liu C, Peng H, Huang T (1995) Crotavirin, a potent platelet aggregation inhibitor purified from the venom of the snake Crotalus viridis. Toxicon 33:1289–1298

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

The authors recognize financial support from the Science and Technology Fund (FONACIT) program (PG-2002000447 and PG-2005000400) of the Ministry of Science and Technology, Caracas, Venezuela.

Author information

Authors and Affiliations

  1. Laboratorio de Investigaciones, Facultad de Farmacia, Universidad Central de Venezuela, Caracas, Venezuela

    Juan C. López-Johnston & Héctor Scannone

  2. Banco Municipal de Sangre de Caracas, Caracas, Venezuela

    Norma de Bosch

  3. Sección de Inmunoquímica, Instituto de Medicina Tropical, Universidad Central de Venezuela, Apartado 47423, Caracas, 1041, Venezuela

    Alexis Rodríguez-Acosta

Authors
  1. Juan C. López-Johnston
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Norma de Bosch
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Héctor Scannone
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Alexis Rodríguez-Acosta
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Alexis Rodríguez-Acosta.

Rights and permissions

Reprints and permissions

About this article

Cite this article

López-Johnston, J.C., de Bosch, N., Scannone, H. et al. Inhibition of collagen, and thrombin-induced platelet aggregation by Lansberg’s hognose pit viper (Porthidium lansbergii hutmanni) venom. J Thromb Thrombolysis 24, 275–282 (2007). https://doi.org/10.1007/s11239-007-0040-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11239-007-0040-x

Keywords

Search

Navigation