Cytotoxic Activities of the Crude Venoms of Macrovipera lebetina lebetina from Cyprus and M. l. obtusa from Turkey (Serpentes: Viperidae) on Human Umbilical Vein Endothelial Cells

Year 2019, Volume: 3 Issue: 2, 110 - 113, 20.12.2019

Naşit İğci , F. Duygu Özel , Mehmet Zülfü Yıldız

https://doi.org/10.31594/commagene.655929

Cited By: 4

Abstract

In this study, we used human umbilical vein endothelial cells (HUVEC) as an in vitro model to compare the cytotoxic activities of the venoms of two Macrovipera lebetina subspecies, M. l. obtusa from southern Anatolia and M. l. lebetina from northern Cyprus. Well-established 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay was preferred to assess the cytotoxicity. Our results showed that venom reduced cell viability both in a time and dose-dependent manner. The cytotoxic effect of M. l. lebetina venom on HUVEC is reported for the first time in the present study.

Keywords

hemorrhage, HUVEC, levantine viper, toxinology

Thanks

We greatly acknowledge and dedicate this article to late Prof. Dr. Bayram GÖÇMEN who passed away tragically on 22 March 2019, for his great contributions to the field herpetology and his invaluable support to the studies related to snake venoms in Turkey by providing venom samples. We also thank Ankara University Biotechnology Institute for providing laboratory access and to Dr. Erkan YILMAZ, Ankara University Biotechnology Institute for providing Human umbilical vein endothelial cells.

Kıbrıs’tan Macrovipera lebetina lebetina ve Türkiye’den M. l. obtusa (Serpentes: Viperidae) Ham Zehirlerinin İnsan Kordon Veni Endotel Hücreleri Üzerindeki Sitotoksik Etkileri

Abstract

Bu çalışmada, Kuzey Kıbrısʼtan Macrovipera lebetina lebetina ve güneydoğu Anadoluʼdan M. l. obtusa olmak üzere iki M. lebetina alt türünün zehirlerinin (venomlarının) sitotoksik aktivitelerini karşılaştırmak için in vitro bir model olarak insan kordon veni endotel hücreleri (HUVEC) kullandık. Sitotoksik etkinin değerlendirilmesinde iyi bilinen bir yöntem olan 3-(4,5-dimethyl-2- hiazolyl)-2,5-diphenyl-2H-tetrazolium bromür (MTT) testi tercih edilmiştir. Çalışmamızın sonuçları her iki zehrin de hücre canlılığını zamana ve doza bağlı olarak azalttığını göstermiştir. M. l. lebetina zehrinin HUVEC üzerindeki sitotoksik etkisi ilk kez bu çalışmada rapor edilmiştir.

Keywords

hemoraji, HUVEC, koca engerek, toksinoloji

References

• Alape-Girón, A., Sanz, L., Escolano, J., Flores-Díaz, M., Madrigal, M., Sasa, M., & Calvete, J.J. (2008). Snake venomics of the Lancehead Pitviper Bothrops asper: Geographic, individual and ontogenetic variations. Journal of Proteome Research, 7(8), 3556-3571.
• Baldo, C., Jamora, C., Yamanouye, N., Zorn, T.M., & Moura-da-Silva, A.M. (2010). Mechanisms of vascular damage by hemorrhagic snake venom metalloproteinases: Tissue distribution and in situ hydrolysis. PLoS Neglected Tropical Diseases, 4(6), e727. https://doi.org/10.1371/journal.pntd.0000727
• Borkow, G., Lomonte, B., Gutiérrez, J.M., & Ovadia, M. (1994). Effect of various Viperidae and Crotalidae Snake Venoms on Endothelial Cells In Vitro. Toxicon, 32, 12, 1689-1695. https://doi.org/10.1016/0041-0101(94)90330-1
• Budak, A., & Göçmen, B. (2008). Herpetoloji. Izmir, Türkiye, Ege Üniversitesi Yayınları, Fen Fakültesi Yayın No: 194, 226 pp.
• Calderon, L.A., Sobrinho, J.C., Zaqueo, K.D., de Moura, A.A., Grabner, A.N., Mazzi, M.V., Marcussi, S., ... Soares, A.M. (2014). Antitumoral activity of snake venom proteins: New trends in cancer therapy. BioMed Reserach International, 2014, 203639, 1-19. http://dx.doi.org/10.1155/2014/203639
• Chippaux J-P. (2006). Snake Venoms and Envenomations. Florida, USA, Krieger Publishing Company, 287 pp.
• Glenn, J.L., Straight, R.C., Wolfe, M.C., & Hardy, D.L. (1983). Geographical variation in Crotalus scutulatus scutulatus (Mojave rattlesnake) venom properties. Toxicon, 21(1), 119-130. https://doi.org/10.1016/0041-0101(83)90055-7
• Göçmen, B., Arıkan, H., Özbel, Y., Mermer, A., & Çiçek, K. (2006). Clinical, Physiological and Serological Observations of a Human Following a Venomous Bite by Macrovipera lebetina lebetina (Reptilia: Serpentes). Acta Parasitologica Turcica, 30(2), 158-162.
• Gutiérrez, J.M., & Rucavado, A. (2000). Snake venom metalloproteinases: Their role in the pathogenesis of local tissue damage. Biochimie, 82, 841-850. https://doi.org/10.1016/S0300-9084(00)01163-9
• Igci, N., & Demiralp, D.O. (2012). A preliminary investigation into the venom proteome of Macrovipera lebetina obtusa (Dwigubsky, 1832) from Southeastern Anatolia by MALDI-TOF mass spectrometry and comparison of venom protein profiles with Macrovipera lebetina lebetina (Linnaeus, 1758) from Cyprus by 2DPAGE. Archives of Toxicology, 86, 441-451. https://doi: 10.1007/s00204-011-0763-5
• Jahromi, H.E., Mirakabadi, A.Z., & Kamalzadeh, M. (2016). Evaluation of Iranian snake “Macrovipera lebetina” venom cytotoxicity in kidney cell line HEK-293. Asia Pacific Journal of Medical Toxicology, 5(2), 49-54.
• Kakanj, M., Ghazi-Khansari, M., Mirakabadi, A.Z., Daraei, B., & Vatanpour, H. (2015). Cytotoxic effect of Iranian Vipera lebetina snake venom on HUVEC cells. Iranian Journal of Pharmaceutical Research, 14, 109-114.
• Mackessy, S.P. (2010). The field of reptile toxinology: Snakes, lizards, and their venoms. In S.P. Mackessy (Ed.), (pp. 3-23). Handbook of Venoms and Toxins of Reptiles. Florida, USA, CRC Press Taylor & Francis Group, 528 pp.
• Mallow, D., Ludwig, D., & Nilson, G. (2003). True Vipers: Natural History and Toxinology of Old World Vipers. Florida, USA, Krieger Publishing Company, 359 pp.
• Moreira, L., Gutiérrez, J.M., Borkow, G., & Ovadia, M. (1992). Ultrastructural alterations in mouse capillary blood vessels after experimental injection of venom from the snake Bothrops asper (Terciopelo). Experimental and Molecular Pathology, 57, 124-133. https://doi.org/10.1016/0014-4800(92)90004-U
• Mosmann, T. (1983). Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. Journal of Immunological Methods, 65, 55-63. https://doi.org/10.1016/0022-1759(83)90303-4
• Nalbantsoy, A., Karabay-Yavasoglu, N.U., Sayım, F., Deliloglu-Gurhan, I., Gocmen, B., Arıkan, H., & Yildiz, M.Z. (2012). Determination of in vivo toxicity and in vitro cytotoxicity of venom from the Cypriot blunt-nosed viper Macrovipera lebetina lebetina and antivenom production. The Journal of Venomous Animals and Toxins including Tropical Diseases, 18(2), 208-216. http://dx.doi.org/10.1590/S1678-91992012000200011
• Oghalaie, A., Kazemi-Lomedasht, F., Zareinejad, M.R., & Shahbazzadeh, D. (2017). Antiadhesive and cytotoxic effect of Iranian Vipera lebetina snake venom on lung epithelial cancer cells. Journal of Family Medicine and Primary Care, 6, 780-783. https://doi.org/10.4103/jfmpc.jfmpc_208_17
• Ozen, M.O., İğci, N., Yalçin, H.T., Goçmen, B., & Nalbantsoy, A. (2015). Screening of cytotoxic and antimicrobial activity potential of Anatolian Macrovipera lebetina obtusa (Ophidia: Viperidae) crude venom. Frontiers in Life Science, 8(4), 363-370. https://doi.org/10.1080/21553769.2015.1055862
• Panfoli, I., Calzia, D., Ravera, S., & Morelli, A. (2010). Inhibition of Hemorragic Snake Venom Components: Old and New Approaches. Toxins, 2, 417-427. https://doi.org/10.3390/toxins2040417
• Samel, M., Trummal, K., Siigur, E., & Siigur, J. (2012). Effect of HUVEC apoptosis inducing proteinase from Vipera lebetina venom (VLAIP) on viability of cancer cells and on platelet aggregation. Toxicon, 60, 648-655. https://doi.org/10.1016/j.toxicon.2012.03.023
• Sanz, L., Ayvazyan, N., & Calvete, J.J. (2008). Snake venomics of the Armenian mountain vipers Macrovipera lebetina obtusa and Vipera raddei. Journal of Proteomics, 71, 198-209. https://doi.org/10.1016/j.jprot.2008.05.003
• Sarıkaya, B., Yıldız, M.Z., & Sezen, G. (2017). The herpetofauna of Adana province (Turkey). Commagene Journal of Biology, 1(1), 1-11. https://doi.org/10.31594/commagene.391784
• Siigur, J., Aaspõllu, A., & Siigur E. (2019). Biochemistry and pharmacology of proteins and peptides purified from the venoms of the snakes Macrovipera lebetina subspecies. Toxicon, 158, 16-32. https://doi.org/10.1016/j.toxicon.2018.11.294
• Stümpel, N., & Joger, U. (2009). Recent advances in phylogeny and taxonomy of near and Middle Eastern Vipers‒an update. ZooKeys, 39, 179-191. https://doi.org/10.3897/zookeys.31.261
• Süzergöz, F., İğci, N., Çavus, C., Yıldız, M.Z., Coşkun, M.B., & Göçmen, B. (2016). In vitro cytotoxic and proapoptotic activities of Anatolian Macrovipera lebetina obtusa (Dwigubski, 1832) crude venom on cultured K562 human chronic myelogenous leukemia cells. UHOD International Journal of Hematology and Oncology, 26(1), 37-46. https://doi.org/10.4999/uhod.161104
• Trummal, K., Tõnismagi, K., Siigur, E., Aaspõllu, A., Lopp, A., Sillat, T., Saat, R., ... Siigur, J. (2005). A novel metalloprotease from Vipera lebetina venom induces human endothelial cell apoptosis. Toxicon, 46(1), 46-61. https://doi.org/10.1016/j.toxicon.2005.03.008
• Yücel Ağan, A.F., & Hayretdağ, S. (2019). The effects of Macrovipera lebetina venom on mice. Toxin Reviews, 38(2), 87-92. https://doi.org/10.1080/15569543.2017.1419266
• Williams, D., Gutiérrez J.M., Harrison, R., Warrell, D.A., White, J., Winkel, K.D., & Gopalakrishnakone, P. (2010). The Global Snakebite Initiative: an antidote for snake bite. Lancet, 375, 89-91. https://doi.org/10.1016/s0140-6736(09)61159-4