Hemolytic activity of skin secretions of amphibians that inhabit the Ukraine territory

Y. Kyriachenko, O. Oskyrko, I. Udovychenko, T. Halenova
Taras Shevchenko National University of Kyiv, Kyiv; Taras Shevchenko National University of Kyiv, Kyiv; Taras Shevchenko National University of Kyiv, Kyiv; Taras Shevchenko National University of Kyiv, Kyiv


Secretions derived from amphibian skin glands serve as a potential reservoir of various valuable active molecules. Currently, the
multiple substances with diverse therapeutic activities among the components of glandular secretions of different species of
amphibians have been found. It has been proven that they have antibacterial, antifungal, antiprotozoal, antidiabetic, antineoplastic,
analgesic, and sleep-inducing properties. Taking this into consideration, to get the basic knowledge about the properties of the
components of skin secretions of some Anura species that inhabit the territory of Ukraine is crucial for further investigation of the
most potential ones. The red blood cell hemolysis assay is a prevalent test to study the cytotoxicity of studied samples. The aim of
the present study was to analyze the hemolytic activity of skin secretions of Bombina bombina, Bombina variegata, Bufotes viridis,
Rana temporaria, Pelophylax ridibundus, and Pelobates fuscus, and to obtain the primary data on the possible mechanism of their
toxicological action on the blood cells membranes. The skin secretions of six amphibian species mentioned above were incubated
with erythrocyte suspension in different concentrations. Eminently active B.variegata skin secretions, having the HD HD50 value at
0.5 µg/ml, were taken for the subsequent researches, where the effects of osmotic protectants, divalent cations, antioxidants,
chelating agent, and serine protease inhibitor on the cell lysis ability of B. variegata skin secretions was studied. All studied cations
inhibited the hemolytic activity of B. variegata secretions in a dose-depend manner. While the serine protease inhibitor,
phenylmethylsulfonyl fluoride (PMSF), markedly decreased the hemolytic activity of studied skin secretions. We can assume that the
bioactive peptides in these skin secretions have an enzymatic mechanism of action.


biologically active molecules, amphibians, hemolytic activity, skin gland secretions

Full Text:



Gomes A., Giri B., Saha A. [et al.]. Bioactive molecules from amphibian skin: Their biological activities with reference to therapeutic potentials for possible drug development. Indian Journal of Experimental Biology. 2007; 45:579-593.

Clarke B. T. The natural history of amphibian skin secretions, their normal functioning and potential medical applications. Biol Rev CambPhilos Soc. 1997; 72(3):365-379.

Lu C. X., Nan K. J., Lei Y. Agents from amphibians with anticancer properties. Anticancer Drugs. 2008; 19(10):931-939. doi: 10.1097/CAD.0b013e3283139100.

Conlon J. M., Kolodziejek J., Nowotny N. Antimicrobial peptides from the skins of North American frogs. https://doi.org/10.1016/j.bbamem.2008.09.

García-Sáez A. J., Buschhorn S. B., Keller H., Anderluh G., Simons K., Schwille P. Oligomerization and pore formation by equinatoxin II inhibit endocytosis and lead to plasma membrane reorganization. J Biol Chem. 2011;19(43):37768–37777.

Savva C. G., Fernandes da Costa S. P., Bokori-Brown M. [et al.]. Molecular architecture and functional analysis of NetB, a pore-forming toxin from Clostridium perfringens. J. Biol Chem. 2013;19(5):3512–3522.

Bradford M. M. A rapid and sensitive for the quantitation of microgram quantitites of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72: 248-254; 1976.

Conlon J. M., Kolodziejek J., Nowotny N. Antimicrobial peptides from ranid frogs: Taxonomic and phylogenetic markers and a potential source of new therapeutic agents. Biochim. Biophys. Acta. 2004;1696:1–14.

You D., Hong J., Rong M. [et al.]. The first gene-encoded amphibian neurotoxin. J. Biol. Chem. 2009;284:22079–22086.

Bevins C. L., Zasloff M. Peptides from frog skin. Annu. Rev. Biochem. 1990;59:395–414.

Daly J. W., Spande T. F., Garraffo H. M. Alkaloids from amphibian

skin: A tabulation of over eight-hundred compounds. J. Nat. Prod.


Mills J. W., Prum B. E. Morphology of the exocrine glands of the frog skin. Am. J. Anat. 1984;171:91–106.

Giovannini M. G., Poulter L., Gibson B. W., Williams D. H. Biosynthesis and degradation of peptides derived from Xenopus laevis prohormones. Biochem J. 1987; 243:113-120.

Darby N. J., Smyth D. G. Characterization of a proteolytic enzyme in the skin secretions of Xenopus laevis. Biochem. Biophys. Res. Commun.1988; 153:1193-1200.

NikolaievaI., Dudkina Yu., Oliinyk D. [et al.]. Amphibian skin secretions: a potential source of proteolytic enzymes. Biotechnologia Acta. 2018; 11(5):42-48.

Jilek A., Mollay Ch., Tippelt Ch. [et al.]. Biosynthesis of a D-amino acid in peptide linkage by an enzyme from frog skin secretions. PNAS. 2005; 102(12):4235-4239; https://doi.org/10.1073/pnas.0500789102.

Received: 16.01.2020

Received a revised version: 17.02.2020

Signed for the press: 17.02.2020

DOI: http://dx.doi.org/10.17721/1728_2748.2020.80.6-10


  • There are currently no refbacks.

Лицензия Creative Commons
This journal is available according to the Creative Commons License «Attribution» («Атрибуція») 4.0 Global (CC-BY).