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Honey Bee Venom (Apis mellifera) Contains Anticoagulation Factors and Increases the Blood-clotting Time

  • Zolfagharian, Hossein (Department of Venomous Animals and Antivenom Production, Razi Vaccine and Serum Research Institute) ;
  • Mohajeri, Mohammad (Department of Toxicology, School of Pharmacy, Shahreza Branch, Islamic Azad University) ;
  • Babaie, Mahdi (Young Researchers and Elites Club, Science and Research Branch, Islamic Azad University)
  • Received : 2015.09.26
  • Accepted : 2015.10.29
  • Published : 2015.12.31

Abstract

Objectives: Bee venom (BV) is a complex mixture of proteins and contains proteins such as phospholipase and melittin, which have an effect on blood clotting and blood clots. The mechanism of action of honey bee venom (HBV, Apis mellifera) on human plasma proteins and its anti-thrombotic effect were studied. The purpose of this study was to investigate the anti-coagulation effect of BV and its effects on blood coagulation and purification. Methods: Crude venom obtained from Apis mellifera was selected. The anti-coagulation factor of the crude venom from this species was purified by using gel filtration chromatography (sephadex G-50), and the molecular weights of the anti-coagulants in this venom estimated by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Blood samples were obtained from 10 rabbits, and the prothrombin time (PT) and the partial thromboplastin time (PTT) tests were conducted. The approximate lethal dose (LD) values of BV were determined. Results: Crude BV increased the blood clotting time. For BV concentrations from 1 to 4 mg/mL, clotting was not observed even at more than 300 seconds, standard deviations $(SDs)={\pm}0.71$; however, clotting was observed in the control group 13.8 s, $SDs={\pm}0.52$. Thus, BV can be considered as containing anti-coagulation factors. Crude BV is composed 4 protein bands with molecular weights of 3, 15, 20 and 41 kilodalton (kDa), respectively. The $LD_{50}$ of the crude BV was found to be $177.8{\mu}g/mouse$. Conclusion: BV contains anti-coagulation factors. The fraction extracted from the Iranian bees contains proteins that are similar to anti-coagulation proteins, such as phospholipase $A_2(PLA_2)$ and melittin, and that can increase the blood clotting times in vitro.

Keywords

References

  1. Son DJ, Lee JW, Lee YH, Song HS, Lee CK, Hong JT. Therapeutic application of anti-arthritis, pain-releasing and anti-cancer effects of bee venom and its constituent compounds. Pharmacol Ther. 2007;115(2):246-70. https://doi.org/10.1016/j.pharmthera.2007.04.004
  2. Habermann E, Jentsch J. Uber die struktur des toxischen bienengiftpeptids melittin und deren beziehung zur pharmakologischen wirkung. Naunyn Schmiedebergs Arch Pharmacol. 1966;253(1):40-1.
  3. Habermann E, Zeuner G. Comparative studies of native and synthetic melittins. Naunyn Schmiedebergs Arch Pharmakol. 1971;270(1):1-9. https://doi.org/10.1007/BF00997294
  4. Shkenderov S, Ivanov T. Pcelni produkti, the bee products (in Bulgarian). Bulgaria: Zemizdat; 1983. 1-238 p.
  5. Kim JI, Yang EJ, Lee MS, Kim YS, Huh Y, Cho IH, et al. Bee venom reduces neuroinflammation in the MPTP-induced model of Parkinson's disease. Int J Neurosci. 2011;121(4):209-17. https://doi.org/10.3109/00207454.2010.548613
  6. Munstedt K, Bogdanov S. Bee products and their potential use in modern medicine. JAAS. 2009;1(3):57-63.
  7. Lee SM, Yang EJ, Choi SM, Kim SH, Baek MG, Jiang JH. Effects of bee venom on glutamate-induced toxicity in neuronal and glial cells. Evid Based Complement Alternat Med. 2012;2012:ID368196.
  8. Orslic N. Bee venom in cancer therapy. Cancer Metastasis Rev. 2012;31(1-2):173-94. https://doi.org/10.1007/s10555-011-9339-3
  9. Mcdonald JA, Li FP, Mehta CR. Cancer mortality among beekeepers. J Occup Med. 1979;21(12):811-3.
  10. Kim JM, Jeon HJ, Kim HJ, Cho CK, Yoo HS. Bee venom pharmacopuncture: an effective treatment for complex regional pain syndrome. J Pharmacopuncture. 2014;17(4):66-9. https://doi.org/10.3831/KPI.2014.17.039
  11. Park JW, Jeon JH, Yoon J, Jung TY, Kwon KR, Cho CK, et al. Effects of sweet bee venom pharmacopuncture treatment for chemotherapy-induced peripheral neuropathy: a case series. Integr Cancer Ther. 2012;11(2):166-71. https://doi.org/10.1177/1534735411413265
  12. Abdu Al-Samie MA. Studies on bee venom and its medical uses. IJOART. 2012;1(2):1-15.
  13. Rizzo F, Papasouliotis K, Crawford E, Dodkin S, Cue S. Measurement of prothrombin time (PT) and activated partial thromboplastin time (APTT) on canine citrated plasma samples following different storage conditions. Res Vet Sci. 2008;85(1):166-70. https://doi.org/10.1016/j.rvsc.2007.10.001
  14. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951;193(1):265-75.
  15. Meier J, Theakston RGD. Approximate LD50 determination of snake venoms using eight to ten experimental animals. Toxicon. 1986;24(4):395-401. https://doi.org/10.1016/0041-0101(86)90199-6
  16. Maulet Y, Brodbeck U, Fulpius BW. Purification from bee venom of melittin devoid of phospholipase A2 contamination. Anal Biochem. 1982;127(1):61-7. https://doi.org/10.1016/0003-2697(82)90144-0
  17. Babaie M, Zolfagharian H, Salmanizadeh H, Mirakabadi AZ, Alizadeh H. Isolation and partial purification of anti-coagulant fractions from the venom of the Iranian snake Echis carinatus. Acta Biochim Pol. 2013;60(1):17-20.
  18. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227(5259):680-5. https://doi.org/10.1038/227680a0
  19. Moreno M, Giralt E. Three valuable peptides from bee and wasp venoms for therapeutic and biotechnological use: melittin, apamin and mastoparan. Toxins. 2015;7(4):1126-50. https://doi.org/10.3390/toxins7041126
  20. Schmidt JO, Blum MS, Overal WL. Comparative enzymology of venoms from stinging hymenoptera. Toxicon. 1986;24(9):907-21. https://doi.org/10.1016/0041-0101(86)90091-7
  21. Shipolini RA, Callewaert GL, Cottrell RG, Vernon CA. The amino-acid sequence and carbohydrate content of phospholipase A2 from bee venom. Eur J Biochem. 1974;48(2):465-76. https://doi.org/10.1111/j.1432-1033.1974.tb03787.x
  22. Hoffman DR, Wood CL. Allergens in hymenoptera venom XI: isolation of protein allergens from Vespula maculifrons (yellow jacket) venom. J Allergy Clin Immunol. 1984;74(1):93-103. https://doi.org/10.1016/0091-6749(84)90094-0
  23. Hoffman DR. Allergens in hymenoptera venom XIII: Isolation and purification of protein components from three species of vespid venoms. J Allergy Clin Immunol. 1985;75(5):599-605. https://doi.org/10.1016/0091-6749(85)90036-3
  24. Hoffman DR, Dove DE, Jacobson RS. Allergens in hymenoptera venom XX: isolation of four allergens from imported fire ant (Solenopsis invicta) venom. J Allergy Clin Immunol. 1988;82(5):818-27. https://doi.org/10.1016/0091-6749(88)90084-X
  25. Edstrom A. Venomous and poisonous animals. Malabar: Krieger Publishing Company; 1992. p. 210-20.
  26. Habermann E. Bee and wasp venoms. Science. 1972;177(4046):314-22. https://doi.org/10.1126/science.177.4046.314
  27. Owen MD, Pfaff LA. Melittin synthesis in the venom system of the honey bee (Apis mellifera L.). Toxicon. 1995;33(9):1181-8. https://doi.org/10.1016/0041-0101(95)00054-P
  28. Benton AW, Morse RA. Venom toxicity and proteins of the genus Apis. J Api Res. 1968;7(3):113-8. https://doi.org/10.1080/00218839.1968.11100200

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