Evaluation of the biological activity of venom extracts from the toads Rhinella marina and Rhaebo guttatus on peripheral blood monocytes from healthy human subjects
DOI:
https://doi.org/10.36560/19320262222Palavras-chave:
Rhinella marina, Rhaebo guttatus, human monocytes, cytotoxicity, immunomodulationResumo
Skin secretions from toads of the Bufonidae family contain immunomodulatory and antioxidant substances and have pharmaceutical potential for treating cancer, degenerative diseases, and cardiovascular conditions. Previous studies have demonstrated that the methanolic extracts of Rhinella marina and Rhaebo guttatus have an immunomodulatory effect on murine macrophages, stimulating the production of pro-inflammatory cytokines and hydrogen peroxide. However, little is known about its effect on human monocytes. Therefore, the aim of this study was to evaluate the in vitro effect of these extracts on the function of human peripheral blood monocytes by assessing cytotoxicity and the production of nitric oxide and hydrogen peroxide. The amphibians were collected in Mato Grosso, Brazil. In the laboratory, their paratoid glands were compressed to extract the venom. Voucher specimens were deposited in the Biological Collection of the Southern Amazon (Sinop/MT). To obtain monocytes, peripheral blood samples were collected from 16 healthy individuals (aged 30–45 years) who had not used any class of medication for the previous 15 days. The monocytes were cultured (5% CO2 at 37°C) with different extract concentrations for 18 h (for cytotoxicity assessment) or 24 h (for activity assessment). For R. marina venom, the most toxic concentration was 1000 µL mL-1, whereas non-toxic concentrations were 62.5 µL mL-1, 15.63 µL mL-1, and 7.8 µL mL-1. For R. guttatus, concentrations ranging from 1000 µL mL-1 to 31.25 µL mL-1 were the most toxic, and the non-toxic concentrations were 15.63 µL mL-1 and 7.8 µL mL-1. The cells did not produce nitric oxide or hydrogen peroxide in response to stimulation with the extracts. We conclude that extracts from the skin secretions of R. marina and R. guttatus exhibit a concentration-dependent cytotoxic effect on human monocytes but do not modulate the cells’ in vitro ability to produce nitric oxide and hydrogen peroxide.
Referências
ABDELFATAH, S.; LU, X.; SCHMEDA-HIRSCHMANN, G.; EFFERTH, T. Cytotoxicity and antimitotic activity of Rhinella schneideri and Rhinella marina venoms. Journal of Ethnopharmacology, 242: 112049, 2019. doi:10.1016/j.jep.2019.112049.
BALDO, M.A.; CUNHA, A.O.S.; GODOY, L.D.; LIBERATO, J.L.; YONEDA, J.S.; FORNARI-BALDO, E.C.; CIANCAGLINI, P.; SANTOS, W.F.; ARANTES, E.C. Assessment of neuropharmacological potential of low molecular weight components extracted from Rhinella schneideri toad poison. The Journal of Venomous Animals and Toxins including Tropical Diseases,25: e148418, 2019. doi:10.1590/1678-9199-JVATITD-1484-18.
BANFI, F.F.; GUEDES, K.S.; ANDRIGHETTI, C.R.; AGUIAR, A.C.; DEBIASI, B.W.; NORONHA, J.C.; RODRIGUES, D.J.; VIEIRA JÚNIOR, G.M.; SANCHEZ, B.A.M. Antiplasmodial and cytotoxic activities of toad venoms from Southern Amazon, Brazil, Brasil. Korean Journal of Parasitology, 54: 415-421, 2016. doi:10.3347/kjp.2016.54.4.415.
CAO, Y.; SONG, Y.U.; AN, N.; ZENG, S.; WANG, D.; YU, L.; ZHU, T.; ZHANG, T.; CUI, J.; ZHOU, C.; DENG, X. The effects of telocinobufagin isolated from Chan Su on the activation and cytokine secretion of immunocytoes in vitro. Fundamental & Clinical Pharmacology, 23: 457-464, 2009. doi:10.1111/j.1472-8206.2009.00696.x.
CARVALHO, D.C.M.; CAVALCANTE-SILVA, L.H.A.; LIMA, E.A.; GALVÃO, J.G.F.M.; ALVES, A.K.A.; FEIJÓ, P.R.O.; QUINTAS, L.E.M.; RODRIGUES-MASCARENHAS, S. Marinobufagenin inhibits neutrophil migration and proinflammatory cytokines. Journal of Immunology Research, 2019: 1094520, 2019. doi: 10.1155/2019/1094520.
CHEN, Y.; HUANG, W.; YANG, M.; XIN, G.; CUI, W.; XIE, Z.; SILVERSTEIN, R.L. Cardiotonic steroids stimulate macrophage inflammatory responses through a pathway involving CD36, TLR4, and Na/K-ATPase. Arteriosclerosis, Thrombosis, and Vascular Biology, 37: 1462-1469, 2017. doi:10.1161/ATVBAHA.117.309444.
CUNHA FILHO, G.A.; SCHWARTZ, C.A.; RESCK, I.S.; MURTA, M.M.; LEMOS, S.; CASTRO, M.S.; KYAW, C.; PIRES JR, O.R.; LEITE, J.R.S.; BLOCH, C.; SCHWARTZ, E.F. Antimicrobial activity of bufadienolides marinobufagin and telocinobufagin isolated as major components from skin secretion of toad Buforubescens. Toxicon, 45: 777-782, 2005. doi: 10.1016/j.toxicon.2005.01.017.
DENG, L.J.; QI, M.; LI, N.; LEI, Y.H.; ZHANG, D.M.; CHEN, J.X. Natural products and their derivatives: promising modulators of tumor immunotherapy, J Leukoc Biol, 108: 493-508, 2020. doi:10.1002/JLB.3MR0320-444R
DIAS-MELICIO, L.A.; CALVI, S.A.; PERAÇOLI, M.T.S.; SOARES, A.M.V.C. Inhibitory effect of deferoxamine on Paracoccidioides brasiliensis survival in human monocytes: reversal by holotransferrin not by apotransferrin. Rev Inst Med Trop S. Paulo, 47: 263-266, 2005. doi:10.1590/s0036-46652005000500005.
FERREIRA, P.M.P.; LIMA, D.J.B.; DEBIASI, B.W.; SOARES, B.M.; MACHADO, K.C.; NORONHA, J.C.; RODRIGUES, D.J.; SINHORIN, A.P.; PESSOA, C.; VIEIRA JÚNIOR, G.M. Antiproliferative activiry of Rhinella marina and Rhaebo guttatus venom extracts from Southern Amazon. Toxicon, 72: 43-51, 2013. doi:10.1016/j.toxicon.2013.06.009
FILHO, E.S.M.; CHAVES, M.H.; FERREIRA, P.M.P.; PESSOA, C.; LIMA, D.J.B.; MARANHÃO, S.S.A.; RODRIGUES, D.J.; JÚNIOR, G.M.V. Cytotoxicity potencial of chemical constituents isolated and derivatised from Rhinella marina venon. Toxicon, 194: 37-43, 2021. doi:10.1016/j.toxicon.2021.02.006
GARCIA, I.J.P.; OLIVEIRA, G.C.; VALADARES, J.M.M.; BANFI, F.F.; ANDRADE, S.N.; FREITAS, T.R.; FILHO, E.S.M.; SANTOS, H.L.; JÚNIO, G.M.V.; CHAVES, M.H.; RODRIGUES, D.J.; SANCHEZ, B.A.M.; VAROTTI, F.P.; BARBOSA, L.A. New bufadienolides extracted from Rhinella marina inhibit Na,K-ATPase and induce apoptosis by activating caspases 3 and 9 in human breast and ovarian cancer cells. Steroids, 152: 108490, 2019. doi:10.1016/j.steroids.2019.108490.
GOMES, V.J.; NUNES, P.R.; MATIAS, M.L.; RIBEIRO, V.R.; DEVIDES, A.C.; BANNWART-CASTRO, C.F.; ROMAGNOLI, G.G.; PERAÇOLI, J.C.; PERAÇOLI, M.T.S.; ROMAO-VEIGA, M. Silibinin induces in vitro M2-like phenotype polarization in monocytes from preeclamptic women. International Immunopharmacology, 89: 107062, 2020. doi:10.1016/j.intimp.2020.107062.
GREEN, L.C.; DE LUZURIAGA, K.R.; WAGNER, D.A.; RAND, W.; ISTFAN, N.; YOUNG, V.R.; TANNENBAUM, S.R. Nitrate biosynthesis in man. Proceedings of the National Academy of Sciences, 78: 7764-7768, 1981. doi:10.1073/pnas.78.12.7764.
HENSON, P.M.; JOHNSON, H.B.; SPIEGELBERG, H.L. The release of granule enzymes from human neutrophils stimulated by aggregated immunoglobulins of different classes and subclasses. J Immunol, 109:1182-92, 1972. https://pubmed.ncbi.nlm.nih.gov/4628704/.
IBARRA-VEGA, R.; GALVAN-HERNÁNDEZ, A.R.; SALAZAR-MONGE, H.; ZATARAÍN-PALACIOS, R.; GARCÍA-VILLALVAZO, P.E.; ZAVALZA-GALVEZ, D.I.; VALDEZ-VELAZQUEZ, L.L.; JIMÉNEZ-VARGAS, J.M. Antimicrobial compounds from skin secretions of species that belong to the Bufonidae Family. Toxins, 15: 145, 2023. doi:10.3390/toxins15020145.
KERKHOFF, J.; NORONHA, J.C.; BONFILIO, R.; SINHORIN, A.P.; RODRIGUES, D.J.; CHAVES, M.H.; JÚNIOR, G.M.V. Quantification of bufadienolides in the poisons of Rhinela marina and Rhaebo guttatus by HPLC-UV. Toxicon, 119: 311 – 318, 2016. doi: 10.1016/j.toxicon.2016.07.003.
LEAL. A.; KARNOPP, E.; BARRETO, Y.C.; OLIVEIRA, R.S.; ROSA, M.E.; BORGES, B.T.; GOULART, F.L.; SOUZA, V.Q.; LAIKOWSKI, M.M.; MOURA, S.; VINADÉ, L.; ROCHA, J.B.T.; BELO, C.A.D. The insecticidal activity of Rhinella schneideri (Werner, 1894) paratoid secretion in Nauphoeta cinerea Cocroaches. Toxins, 12: 630, 2020. doi:10.3390/toxins12100630
LIU, J.; LILLY, M.N.; SHAPIRO, J.I. Targeting Na/K-ATPase signaling: a new approach to control oxidative stress. Current Pharmacology Des., 24: 359-364, 2018. doi:10.2174/1381612824666180110101052.
LUCISANO, Y.M.; MANTOVANI, B. Lysosomal-enzyme release from polymorphonuclear leukocytes induced by immune-complexes of IgM and IgG. Journal of Immunology, 132: 2015-2020, 1984. https://pubmed.ncbi.nlm.nih.gov/6699405/.
MARTINS, P.R.; SOARES, A.M.V.C.; DOMENEGHINI, A.V.S.P.; GOLIM, M.A.; KANENO, R. Agaricus brasiliensis polysaccharides stimulate human monocytes to capture Candida albicans, express toll-like receptors 2 and 4, and produce pro-inflammatory cytokines. Journal of Venomous Animals and Toxins including Tropical Diseases, 23: 17, 2017. doi:10.1186/s40409-017-0102-2.
MATIAS, M.L.; GOMES, V.J.; ROMAO-VEIGA, M.; RIBEIRO, V.R.; NUNES, P.R.; ROMAGNOLI, G.G.; PERACOLI, J.C.; PERACOLI, M.T.S. Silibinin downregulates the NF-kB pathway and NLRP1/NLRP3 inflammasones in monocytes from pregnant women with preeclampsia. Molecules, 24: 1548, 2019. doi:10.3390/molecules24081548.
MEDEIROS, D.S.S.; REGO, T.B.; SANTOS, A.P.A.; PONTES, A.S.; MOREIRA-DILL, L.S.; MATOS, N.B.; ZULIANI, J.P.; STÁBELI, R.G.; TELES, C.B.G.; SOARES, A.M.; SPEROTTO, A.R.M.; MOURA, D.J.; SAFFI, J.; CALDEIRA, C.A.S.; PIMENTA, D.C.; CALDERON, L.A. Biochemical and biological profile of parotoid secretion of the Amazonian Rhinella marina (Anura: Bufonidae). Biomed Research International, 2019: 2492315, 2019. doi: 10.1155/2019/2492315.
OLIVEIRA, A.F.; CASTOLDI, L.; VIEIRA JR, G.M.; MONÇÃO FILHO, E.S.; CHAVES, M.H.; RODRIQUES, D.J.; SUGUI, M.M. Evaluation of antimutagenic and cytotoxic activity of skin secretion extract of Rhinella marina and Rhaebo guttatus (Anura, Bufonidae). Acta Amazonica, 49: 145 – 151, 2019. doi:10.1590/1809-4392201801751.
OZANSKA, A.; SZYMCZACK, D.; RYBKA, J. Pattern of human monocyte subpopulations in health and disease. Scandinavian Journal of Immunology, 92: e12883, 2020. doi: 10.1111/sji.12883.
PELISSARI, S.N.; SOUZA, E.R.; SINHORIN, V.G.; RODRIGUES, D.D.; CASTOLDI, L.; SINHORIN, A.P. A crude methanolic extract from the parotoid gland secretion of Rhaebo guttatus stimulates the production of reactive species and pro inflammatory cytokines by peritoneal macrophages. World Acad Sci J, 5: 15, 2023. doi:10.3892/wasj.2023.192.
PELISSARI, S.R.N.; SINHORIN, V.D.G.; CASTOLDI, L.; VASCONCELOS, L.G.; RODRIGUES, D.J.; RIBEIRO, E.B.S.; KERKHOFF, J.; SINHORIN, A.P. Methanolic Extract of Rhinella marina Poison: Chemical Composition, Antioxidant and Immunomodulatory Activities. J Braz Chem Soc, 32: 1584-1597, 2021. doi:10.21577/0103-5053.20210057.
PERAÇOLI, M.T.S.; BANNWART, C.F.; CRISTOFALO, R.; BORGES, V.T.M.; COSTA, R.A.A.; WITKIN, S.S.; PERAÇOLI, J.C. Increased reactive oxygen species and tumor necrosis factor-alpha production by monocytes are associated with elevated levels of uric acid in pre-eclamptic women. American Journal of Reproductive Immunlogy, 66: 460-467, 2011. doi:10.1111/j.1600-0897.2011.01016.x.
PICK, E.; MIZEL, D. Rapid microassays for the measurement of superoxide and hydrogen peroxide production by macrophages in culture using an automatic enzyme immunoassay reader. J Immunol Methods, 46: 211-226, 1981. doi:10.1016/0022-1759(81)90138-1.
PRASSAS, I.; DIAMANDIS, E.P. Novel therapeutic applications of cardiac glycosides. Nature Reviews Drug Discovery, 7: 926-935, 2008. doi:10.1038/nrd2682.
RAASCH-FERNANDES, L.D.; BONALDO, S.M.; RODRIGUES, D.J.; FERRARINI, S.R.; VERÇOSA, A.G.A.; OLIVEIRA, D.L. In vitro antimicrobial activity of methanolic extracts from cutaneous secretions of Amazonian amphibians against phytopathogens of agricultural interest. Acta Amazonica, 51: 145-155, 2021. doi:10.1590/1809-4392201904462.
RODRIGUEZ, C.; IBÁÑEZ, R.; OLMEDO, D.A.; NG, M.; SPADAFORA, C.; DURANT-ARCHIBOLD, A.A.; GUTIÉRREZ, M. Anti-Trypanosomal Bufadienolides from the Oocytes of the Toad Rhinella alata (Anura, Bufonidae). Molecules, 29: 196, 2024. doi:10.3390/molecules29010196.
SANTOS, C.V.; KERKHOFF, J.; TOMAZELLE, C.A.; WENCESLAU, C.F.; SINHORIN, A.P.; RODRIGUES, D.J.; CARNEIRO, F.S.; BOMFIM, G.F. Vasoconstrictor and hemodynamic effects of a methanolic extract from Rhinella marina toad poison. Toxicon, 218, 57 – 65, 2022. doi:10.1016/j.toxicon.2022.08.018.
SCHMEDA-HIRSCHMANN, G.; BURGOS-EDWARDS, A.; ARIAS, A.R.; LÓPES-TORRES, C.; PALOMINOS, C.; FUENTES-RETAMAL, S.; HERRERA, Y.; DUBOIS-CAMACHO, K.; URRA, F.A. A Paraguayan toad Rhinella schneideri preparation based on Mbya tradition increases mitochondrial bioenergetics with migrastatic effects dependent on AMPK in breast cancer cells. Journal of Ethnopharmacology, 294: 115344, 2022. doi:10.1016/j.jep.2022.115344.
SHIH, Y.L.; CHOU, J.S.; CHEN, Y.L.; HSUEH, S.C.; CHUNG, H.Y.; LEE, M.H.; CHEN, C.P.; LEE, M.Z.; HOU, H.T.; LU, H.F.; CHEN, K.W.; CHUNG, J.G. Bufalin enhances immune responses in leukemic mice through enhancing phagocytosis of macrophage in vivo. In Vivo, 32:1129-1136, 2018. doi:10.21873/invivo.11355.
SOUSA, L.Q.; MACHADO, K.C.; OLIVEIRA, S.F.C.; ARAÚJO, L.S.; MONÇÃO-FILHO, E.S.; MELO-CAVALCANTE, A.A.C.; VIEIRA-JÚNIO, G.M.; FERREIRA, P.M.P. Bufadienolides from amphibians: A promising source of anticancer prototypes for radical innovation, apoptosis triggering and Na+/K+ - ATPase inhibition. Toxicon, 127: 63-76, 2017. doi:10.1016/j.toxicon.2017.01.004
SOUZA, E.B.R.; JÚNIOR, P.T.S.; VASCONCELOS, L.G.; RODRIGUES, D.J.; SINHORIN, V.D.G.; KERKHOFF, J.; PELISSARI, S.R.N.; SINHORIN, A.P. Comparative study of the chemical profile of the parotoid gland secretions from Rhaebo guttatus from diferente regions of the Brazilian Amazon. Toxicon, 179: 101-106, 2020. doi:10.1016/j.toxicon.2020.03.005.
SOUZA, L.P.F.; SOUZA, M.E.P.; CASTOLDI, L. Natural products: the contribution of research developed by a university in Sinop, Brazil. Scientific Electronic Archives,16: np, 2023. doi:10.36560/16820231769.
SPINELLI, R.; RIETMANN, A.; SANCHIS, I.; SIANO, A.S. Natural multi-target modulators of pathological pathways in Alzheimer’s disease isolated from the Rhinella arenarum skin. Natural Product Research, 36: 3193 – 3197, 2022. doi:10.1080/14786419.2021.1953022.
WANG, D.L.; QI, F.H.; TANG, W.; WANG, F.S. Chemical constituents and bioactivities of the skin of Bufo bufo gargarizans Cantor. Chemistry & Biodiversity, 8: 559–567, 2011. doi:10.1002/cbdv.201000283.
XIE, S.; SPELMINK, L.; CODEMO, M.; SUBRAMANIAN, K.; PÜTSEP, K.; HENRIQUES-NORMARK, B.; OLLIVER, M. Cinobufagin modulates human innate immune responses and triggers antibacterial activity. Plos One, 11: e0160734, 2016. doi:10.1371/journal.pone.0160734.
YU, Y.; WANG, H.; MENG, X.; HAO, L.; FU, Y.; FANG, L.; SHEN, D.; YU, X.; LI, J. Immunomodulatory effects of cinobufagin on murine lymphocytes and macrophages. Evidence-based Complementary and Alternative Medicine, 2015: 835263, 2015. doi: 10.1155/2015/835263.
ZHANG, J.; CHANG, J.; BEG, M.A.; HUANG, W.; ZHAO, Y.; DAI, W.; WU, X.; CUI, W.; PILLAI, S.S.; LAKHANI, H.V.; SODHI, K.; SHAPIRO, J.I.; SAHOO, D.; ZHENG, Z.; SILVERSTEIN, R.L.; CHEN, Y. Na/K-ATPase suppress LPS-induced pro-inflammatory signaling through Lyn. iScience, 25: 104963, 2022. doi: 10.1016/j.isci.2022.104963
Downloads
Publicado
Como Citar
Edição
Seção
Licença
Copyright (c) 2026 Scientific Electronic Archives
Este trabalho está licenciado sob uma licença Creative Commons Attribution 4.0 International License.
A revista se reserva o direito de fazer alterações nas regras originais, na ortografia e na ordem gramatical, a fim de manter o idioma de culto padrão, respeitando, no entanto, o estilo dos autores. Os artigos publicados são de propriedade da revista Scientific Electronic Archives, tornando-se sua reimpressão total ou parcial, sujeitos à autorização expressa da direção da revista. A fonte original da publicação deve ser mantida. Os originais não serão devolvidos aos autores. As opiniões expressas pelos autores dos artigos são de sua exclusiva responsabilidade.
The journal reserves the right to make changes to the original rules, spelling and grammatical order, in order to keep the language of worship default, respecting, however, the style of the authors. Articles published are the property of Scientific Electronic Archives magazine, becoming its total or partial reprint, subject to the express authorization of the direction of the journal. The original source of publication should be retained. The originals will not be returned to the authors. Opinions expressed by authors of articles are solely your responsibility.
This journal uses the License Creative Commons Atribuição 4.0 Internacional.
Accepted 2026-04-30
Published 2026-05-08
