The spermicidal effect of the honeybee Apis mellifera venom and its major component melittin

Document Type : Original Article

Authors

1 Molecular biology research lab., Department of Zoology, Faculty of Science (Assiut), Al-Azhar University, Egypt.

2 Department of Zoology, Faculty of Science, Suez Canal University, Egypt.

3 Department of Obstetrics and Gynaecology, Faculty of Medicine, Cairo University, Egypt.

Abstract

Several spermicide products are currently used containing detergent as the active ingredient. Those molecules showed a bad effect on normal vaginal epithelial cells and microflora. They also increase the possibility of cervical or vaginal infections or ulceration. In this study, the possible effects of crude bee venom (BV) of the honeybee (Apis mellifera) and its common bioactive peptides on human sperm motility were evaluated in vitro. Serially diluted BV and its bioactive peptides at noncytotoxic concentrations were tested for sperm immobilization by Sander-Cramer assay. The sperm viability test and the hypo-osmotic swelling were used to check the vitality and the integrity of the sperm membrane. Sperm morphological alterations were evaluated by a light microscope and scanning electron microscope. Results showed that BV at 3.0 μg/ml concentration was significantly immobilized and killed 100% (1.0 million) of the human sperm within 20 seconds. Similarly, the small peptide MLT showed immediate and complete sperm immobilization at 6.7 µg/ml. Moreover, apamin induced a moderate effect on sperm motility at 10.0 μg/ml, whereas mast cell degranulating peptide showed no effect on sperm motility up to 10.0 μg/ml. In conclusion, the current study indicates that BV and MLT have a spermicidal effect and exert their spermicidal activity through a direct lysis effect on the plasma membranes of sperm heads.

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Main Subjects


[1]           Haub C, Kaneda T. World Population Data Sheet–Population Reference Bureau. 2019. Population Reference Bureau (PRB). 2013.
[2]           Doncel GF. Exploiting common targets in human fertilization and HIV infection: development of novel contraceptive microbicides. Human reproduction update. 2006;12(2):103-17.
[3]           Mprah WK. Perceptions about barriers to sexual and reproductive health information and services among deaf people in Ghana. 2021.
[4]           Singh KK, Parmar S, Tatke PA. Contraceptive efficacy and safety of HerbOshield™ vaginal gel in rats. Contraception. 2012;85(1):122-7.
[5]           D’Cruz OJ, Uckun FM. Gel-microemulsions as vaginal spermicides and intravaginal drug delivery vehicles. Contraception. 2001;64(2):113-23.
[6]           Krebs FC, Miller SR, Catalone BJ, Welsh PA, Malamud D, Howett MK, et al. Sodium dodecyl sulfate and C31G as microbicidal alternatives to nonoxynol 9: comparative sensitivity of primary human vaginal keratinocytes. Antimicrobial Agents and Chemotherapy. 2000;44(7):1954-60.
[7]           D'Cruz OJ, Shih M-J, Yiv SH, Chen C-L, Uckun FM. Synthesis, characterization and preclinical formulation of a dual-action phenyl phosphate derivative of bromo-methoxy zidovudine (compound WHI-07) with potent anti-HIV and spermicidal activities. Molecular human reproduction. 1999;5(5):421-32.
[8]        D'Cruz OJ, Uckun FM. Novel derivatives of phenethyl-5-bromopyridylthiourea and dihydroalkoxybenzyloxopyrimidine are dual-function spermicides with potent anti-human immunodeficiency virus activity. Biology of reproduction. 1999;60(6):1419-28.
[9]           Klebanoff S. Effects of the spermicidal agent nonoxynol-9 on vaginal microbial flora. Journal of Infectious Diseases. 1992;165(1):19-25.
[10]         Martin Jr HL, Richardson BA, Nyange PM, Lavreys L, Hillier SL, Chohan B, et al. Vaginal lactobacilli, microbial flora, and risk of human immunodeficiency virus type 1 and sexually transmitted disease acquisition. Journal of Infectious Diseases. 1999;180(6):1863-8.
[11]         Martinez J, Sasse F, Brönstrup M, Diez J, Meyerhans A. Antiviral drug discovery: broad-spectrum drugs from nature. Natural product reports. 2015;32(1):29-48.
[12]         El-Bitar AM, Sarhan M, Abdel-Rahman MA, Quintero-Hernandez V, Aoki-Utsubo C, Moustafa MA, et al. Smp76, a scorpine-like peptide isolated from the venom of the scorpion Scorpio maurus palmatus, with a potent antiviral activity against hepatitis C virus and dengue virus. International Journal of Peptide Research and Therapeutics. 2020;26(2):811-21.
[13]         Cunha RdS, Vigerelli H, Jared C, Antoniazzi MM, Chaves LB, Silva AdCRd, et al. Synergic effects between ocellatin-F1 and bufotenine on the inhibition of BHK-21 cellular infection by the rabies virus. Journal of Venomous Animals and Toxins including Tropical Diseases. 2015;21.
[14]         Rivero J, de Castro F, Stival A, Magalhães M, Carmo Filho J, Pfrimer I. Mechanisms of virus resistance and antiviral activity of snake venoms. Journal of Venomous Animals and Toxins including Tropical Diseases. 2011;17(4):387-93.
[15]         Vigerelli H, Sciani JM, Jared C, Antoniazzi MM, Caporale GMM, Silva AdCRd, et al. Bufotenine is able to block rabies virus infection in BHK-21 cells. Journal of Venomous Animals and Toxins including Tropical Diseases. 2014;20:02-8.
[16]         Hmed B, Serria HT, Mounir ZK. Scorpion peptides: potential use for new drug development. Journal of toxicology. 2013;2013.
[17]         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.
[18]         Park HJ, Lee SH, Son DJ, Oh KW, Kim KH, Song HS, et al. Antiarthritic effect of bee venom: Inhibition of inflammation mediator generation by suppression of NF‐κB through interaction with the p50 subunit. Arthritis & rheumatism. 2004;50(11):3504-15.
[19]         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. Pharmacology & therapeutics. 2007;115(2):246-70.
[20]         Rasul A, Millimouno FM, Ali Eltayb W, Ali M, Li J, Li X. Pinocembrin: a novel natural compound with versatile pharmacological and biological activities. BioMed research international. 2013;2013.
[21]         Schnitzler P, Neuner A, Nolkemper S, Zundel C, Nowack H, Sensch KH, et al. Antiviral activity and mode of action of propolis extracts and selected compounds. Phytotherapy Research. 2010;24(S1):S20-S8.
[22]         Falco A, Barrajón-Catalán E, Menéndez-Gutiérrez MP, Coll J, Micol V, Estepa A. Melittin-loaded immunoliposomes against viral surface proteins, a new approach to antiviral therapy. Antiviral research. 2013;97(2):218-21.
[23]         Lee J-A, Kim Y-M, Hyun P-M, Jeon J-W, Park J-K, Suh G-H, et al. Honeybee (Apis mellifera) venom reinforces viral clearance during the early stage of infection with porcine reproductive and respiratory syndrome virus through the up-regulation of Th1-specific immune responses. toxins. 2015;7(5):1837-53.
[24]         Uddin MB, Lee B-H, Nikapitiya C, Kim J-H, Kim T-H, Lee H-C, et al. Inhibitory effects of bee venom and its components against viruses in vitro and in vivo. Journal of Microbiology. 2016;54(12):853-66.
[25]         Liu H, Han Y, Fu H, Liu M, Wu J, Chen X, et al. Construction and expression of sTRAIL–melittin combining enhanced anticancer activity with antibacterial activity in Escherichia coli. Applied microbiology and biotechnology. 2013;97(7):2877-84.
[26]         Mataraci E, Dosler S. In vitro activities of antibiotics and antimicrobial cationic peptides alone and in combination against methicillin-resistant Staphylococcus aureus biofilms. Antimicrobial agents and chemotherapy. 2012;56(12):6366-71.
[27]         Billingham M, Morley J, HANSON JM, Shipolini R, Vernon C. An anti-inflammatory peptide from bee venom. Nature. 1973;245(5421):163-4.
[28]         Habermann E. Bee and wasp venoms. Science. 1972;177(4046):314-22.
[29]         Hwang D-S, Kim SK, Bae H. Therapeutic effects of bee venom on immunological and neurological diseases. Toxins. 2015;7(7):2413-21.
[30]         Protocol–BVT L, Blog H, Charts MS, Page V, Extremities SY, vs Remission R, et al. Bee Venom in Cancer Therapy. Cancer Metastasis Rev. 2012;31:173-94.
[31]         Organisation WH. WHO laboratory manual for the examination of human semen and sperm-cervical mucus interaction: Cambridge university press; 1999.
[32]         Das N, Chandran P, Chakraborty S. Potent spermicidal effect of oleanolic acid 3-beta-D-glucuronide, an active principle isolated from the plant Sesbania sesban Merrill. Contraception. 2011;83(2):167-75.
[33]         Green TR, Fellman JH, Wolf DP. Human spermicidal activity of inorganic and organic oxidants. Fertility and sterility. 2001;76(1):157-62.
[34]         Paul CR. Introduction to electromagnetic compatibility: John Wiley & Sons; 2006.
[35]         Nussdorfer P, Cilenšek I, Zorn B, Petrovič D. Adapted methods for scanning electron microscopy (SEM) in assessment of human sperm morphology. Bosnian journal of basic medical sciences. 2018;18(1):43.
[36]         Pucca MB, Cerni FA, Oliveira IS, Jenkins TP, Argemí L, Sørensen CV, et al. Bee updated: Current knowledge on bee venom and bee envenoming therapy. Frontiers in immunology. 2019;10:2090.
[37]         Jo M, Park MH, Kollipara PS, An BJ, Song HS, Han SB, et al. Anti-cancer effect of bee venom toxin and melittin in ovarian cancer cells through induction of death receptors and inhibition of JAK2/STAT3 pathway. Toxicology and applied pharmacology. 2012;258(1):72-81.
[38]         Rady I, Siddiqui IA, Rady M, Mukhtar H. Melittin, a major peptide component of bee venom, and its conjugates in cancer therapy. Cancer letters. 2017;402:16-31.
[39]         Choi KE, Hwang CJ, Gu SM, Park MH, Kim JH, Park JH, et al. Cancer cell growth inhibitory effect of bee venom via increase of death receptor 3 expression and inactivation of NF-kappa B in NSCLC cells. Toxins. 2014;6(8):2210-28.
[40]         Hood JL, Jallouk AP, Campbell N, Ratner L, Wickline SA. Cytolytic nanoparticles attenuate HIV-1 infectivity. Antivir Ther. 2013;18(1):95-103.41][     Sarhan M, El-Bitar AM, Hotta H. Potent virucidal activity of honeybee “Apis mellifera” venom against Hepatitis C Virus. Toxicon. 2020;188:55-64.
[42]         Flesch FM, Gadella BM. Dynamics of the mammalian sperm plasma membrane in the process of fertilization. Biochimica et Biophysica Acta (BBA)-Reviews on Biomembranes. 2000;1469(3):197-235.
[43]         Tapia J, Macias‐Garcia B, Miro‐Moran A, Ortega‐Ferrusola C, Salido G, Pena F, et al. The membrane of the mammalian spermatozoa: much more than an inert envelope. Reproduction in domestic animals. 2012;47:65-75.
[44]         Zairi A, Belaïd A, Gahbiche A, Hani K. Spermicidal activity of dermaseptins. Contraception. 2005;72(6):447-53.
[45]         Escoffier J, Couvet M, De Pomyers H, Ray PF, Seve M, Lambeau G, et al. Snake venoms as a source of compounds modulating sperm physiology: Secreted phospholipases A2 from Oxyuranus scutellatus scutellatus impact sperm motility, acrosome reaction and in vitro fertilization in mice. Biochimie. 2010;92(7):826-36.
[46]         Parodi J, Navarrete P, Marconi M, Gutiérrez RS, Martínez‐Torres A, Mejías FR. Tetraethylammonium-sensitive K+ current in the bovine spermatozoa and its blocking by the venom of the Chilean Latrodectus mactans. Systems biology in reproductive medicine. 2010;56(1):37-43.
[47]         DeGrado W, Musso G, Lieber M, Kaiser E, Kezdy F. Kinetics and mechanism of hemolysis induced by melittin and by a synthetic melittin analogue. Biophysical journal. 1982;37(1):329-38.
[48]         Terwilliger TC, Weissman L, Eisenberg D. The structure of melittin in the form I crystals and its implication for melittin's lytic and surface activities. Biophysical journal. 1982;37(1):353-61.
[49]         Dufourc EJ, Smith IC, Dufourcq J. Molecular details of melittin-induced lysis of phospholipid membranes as revealed by deuterium and phosphorus NMR. Biochemistry. 1986;25(21):6448-55.
[50]         Tosteson M, Holmes S, Razin M, Tosteson D. Melittin lysis of red cells. The Journal of membrane biology. 1985;87(1):35-44.
[51]         Cuppoletti J, Blumenthal KM, Malinowska DH. Melittin inhibition of the gastric (H++ K+) ATPase and photoaffinity labeling with [125I] azidosalicylyl melittin. Archives of biochemistry and biophysics. 1989;275(1):263-70.
[52]         Cuppoletti J, Abbott AJ. Interaction of melittin with the (Na++ K+) ATPase: evidence for a melittin-induced conformational change. Archives of biochemistry and biophysics. 1990;283(2):249-57.
[53]         Clague MJ, Cherry RJ. Comparison of p25 presequence peptide and melittin. Red blood cell haemolysis and band 3 aggregation. Biochemical Journal. 1988;252(3):791-4.
[54]         Voss J, Birmachu W, Hussey DM, Thomas DD. Effects of melittin on molecular dynamics and calcium-atpase activity in sarcoplasmic reticulum membranes: Time-resolved optical anisotropy. Biochemistry. 1991;30(30):7498-506.
[55]         Kuchinka E, Seelig J. Interaction of melittin with phosphatidylcholine membranes. Binding isotherm and lipid head-group conformation. Biochemistry. 1989;28(10):4216-21.
[56]         Ekici BB, Aksoy UT. Prediction of building energy needs in early stage of design by using ANFIS. Expert Systems with Applications. 2011;38(5):5352-8.
[57]         Am-In N, Kirkwood R, Techakumphu M, Tantasuparuk W. Lipid profiles of sperm and seminal plasma from boars having normal or low sperm motility. Theriogenology. 2011;75(5):897-903.
[58]         Beer-Ljubić B, Aladrović J, Marenjak T, Laškaj R, Majić-Balić I, Milinković-Tur S. Cholesterol concentration in seminal plasma as a predictive tool for quality semen evaluation. Theriogenology. 2009;72(8):1132-40.
[59]         Cerolini S, Maldjian A, Surai P, Noble R. Viability, susceptibility to peroxidation and fatty acid composition of boar semen during liquid storage. Animal Reproduction Science. 2000;58(1-2):99-111.