Vellozia flavicans Mart. ex Schult. hydroalcoholic extract inhibits the neuromuscular blockade induced by Bothrops jararacussu venom

FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOBackground: Snakebite is a significant public health issue in tropical countries. In Brazil, some of the most common snake envenomations are from Bothrops. Bothrops bites trigger local and systemic effects including edema, pain, erythema, cyanosis, infections, and necrosis. Vellozia flavicans is a plant from the Brazilian " cerrado" (savanna) that is popularly used as an anti-inflammatory medicine. Since inflammation develops quickly after Bothrops bites, which can lead to infection, the aim of the present study was to observe possible anti-snake venom and antimicrobial activities of V. flavicans (Vf). Methods: The chromatographic profile of the main constituents from the Vf leaf hydroalcoholic extract was obtained by thin-layer chromatography (TLC). The anti-snake venom activity was measured by Vf's ability to neutralize the in vitro neuromuscular blockade caused by Bothrops jararacussu venom (Bjssu) in a mouse phrenic nerve-diaphragm model (PND). After a 20 min incubation, preparations of PND were added to Tyrode's solution (control); Vf (0.2, 0.5, 1, and 2 mg/mL); 40 μg/mL Bjssu; pre-incubation for 30 min with Bjssu and 1 mg/mL Vf; and a Bjssu pretreated preparation (for 10 min) followed by 1 mg/mL Vf. Myographic recording was performed, and the contractile responses were recorded. The antimicrobial activity (minimum inhibitory concentration [MIC] and minimum bactericidal concentration [MBC]) was obtained for Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, and Enterococcus faecalis, using gentamicin and vancomycin as positive controls. Results: TLC analysis yielded several compounds from Vf, such as flavonoids (quercetin) and phenolic acids (chlorogenic acid). Bjssu completely blocked the contractile responses of PND preparations, while Vf preserved 97% (±10%) of the contractile responses when incubated with Bjssu. In the PND pretreated with Bjssu, Vf was able to inhibit the neuromuscular blockade progress. MIC and MBC of Vf ranged from 2.5 to 5.0 mg/mL for P. aeruginosa and S. aureus strains, while no antimicrobial activity was observed for E. coli and E. faecalis.Conclusions: The hydroalcoholic extract from Vf leaves was able to neutralize and decrease the in vitro neuromuscular blockade caused by Bjssu. However, it did not show significant antimicrobial activity against the tested bacteria. © 2014 Tribuiani et al.; licensee BioMed Central Ltd.Snakebite is a significant public health issue in tropical countries. In Brazil, some of the most common snake envenomations are from Bothrops. Bothrops bites trigger local and systemic effects including edema, pain, erythema, cyanosis, infections, and necrosis. Vellozia flavicans is a plant from the Brazilian "cerrado" (savanna) that is popularly used as an anti-inflammatory medicine. Since inflammation develops quickly after Bothrops bites, which can lead to infection, the aim of the present study was to observe possible anti-snake venom and antimicrobial activities of V. flavicans (Vf). The chromatographic profile of the main constituents from the Vf leaf hydroalcoholic extract was obtained by thin-layer chromatography (TLC). The anti-snake venom activity was measured by Vf's ability to neutralize the in vitro neuromuscular blockade caused by Bothrops jararacussu venom (Bjssu) in a mouse phrenic nerve-diaphragm model (PND). After a 20 min incubation, preparations of PND were added to Tyrode's solution (control); Vf (0.2, 0.5, 1, and 2 mg/mL); 40 μg/mL Bjssu; pre-incubation for 30 min with Bjssu and 1 mg/mL Vf; and a Bjssu pretreated preparation (for 10 min) followed by 1 mg/mL Vf. Myographic recording was performed, and the contractile responses were recorded. The antimicrobial activity (minimum inhibitory concentration [MIC] and minimum bactericidal concentration [MBC]) was obtained for Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, and Enterococcus faecalis, using gentamicin and vancomycin as positive controls. TLC analysis yielded several compounds from Vf, such as flavonoids (quercetin) and phenolic acids (chlorogenic acid). Bjssu completely blocked the contractile responses of PND preparations, while Vf preserved 97% (±10%) of the contractile responses when incubated with Bjssu. In the PND pretreated with Bjssu, Vf was able to inhibit the neuromuscular blockade progress. MIC and MBC of Vf ranged from 2.5 to 5.0 mg/mL for P. aeruginosa and S. aureus strains, while no antimicrobial activity was observed for E. coli and E. faecalis. The hydroalcoholic extract from Vf leaves was able to neutralize and decrease the in vitro neuromuscular blockade caused by Bjssu. However, it did not show significant antimicrobial activity against the tested bacteria14FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO2004/09705-8; 2007/53883-6; 2008/52643-4; 2008/11005-5; 2012/08271-0Neglected tropical diseases: Snakebite, , http://www.who.int/bloodproducts/animal_sera/Rabies.pdf?ua=1, WHO - World Health Organization(2009) Textos Básicos de Saúde (Cadernos de Atenção Básican. 22), , Health surveillance:zoonoses, Brasil. Ministério da SaúdeTeixeira, R., Forma grave do acidente por ofídios da sub-família Crotalinae (1979) An Acad Med Bahia, 2, pp. 109-135Vellard, J.A., Serpentes venenosas (1945) Terapêutica Clínica, pp. 265-273. , Buenos Aires: Libreria y Editorial 'El Ateneo, Cardini C, Beretervide JJAlves, E., (1956) Medicina de Urgência, pp. 1052-1055. , Rio de Janeiro: Livraria AtheneuBrazil, V., (1901) Do envenenamento ophidico e seu tratamento, pp. 31-55. , Colletanea dos trabalhos Instituto ButantanRodrigues-Simioni, L., Borgese, N., Ceccarelli, B., The effects of Bothrops jararacussu venom and its components on frog nerve-muscle preparation (1983) Neuroscience, 10, pp. 475-489Souza, C.D., Felfili, J.M., The utilization of medicinal plants in the region of Alto Paraíso of Goiás, GO, Brazil (2006) Acta Bot Bras, 20, pp. 133-142Stannard, B.L., (1995) Flora of the Pico das Almas: Chapada da Diamantina - Bahia, Brazil, pp. 43-78. , Great Britain: Whitstable Litho LtdBranco, A., Pinto, A.C., Braz Filho, R., Chemical constituents from Vellozia graminifolia (Velloziaceae) (2004) An Acad Bras Cienc, 76, pp. 505-518Harborne, J.B., Williams, C.A., Greenham, J., Eagles, J., Variations in the lipophilic and vacuola flavonoids of the genus Vellozia (1994) Phytochemistry, 35, pp. 1475-1480Williams, C.A., Harborne, J.B., Greenham, J., Eagles, J., Differences in flavonoids patterns between genera within the Velloziaceae (1994) Phytochemistry, 36, pp. 931-940Pinto, A.C., Scofield, T.C.V., Braz-Filho, R., Two new diterpenes with a rosane skeleton from Velloziaceae (1983) Tetrahedron Lett, 24, pp. 5043-5046Pinto, A.C., Queiroz, P.P.S., Garcez, W., Diterpenes from Vellozia bicolor (1991) J Braz Chem Soc, 2, pp. 25-30Pinto, A.C., Rezende, C.M., Antunes, O.A.C., Correia, C.R.D., Three isomeric diterpenes from Vellozia flavicans (1996) Phytochemistry, 42, pp. 767-769Peixoto, E.M., Pinchin, R., Pinto, A.C., Constituintes químicos de Vellozia piresiana (1979) Ciênc Cult, 32, pp. 125-127Barnes, R.A., Pereira, A.L., Scofield, T.C.V., Braz-Filho, R., Pinto, A.C., A new triterpene from Vellozia compacta (1984) Chem Pharm Bull, 32, pp. 3674-3677Dharmappa, K.K., Kumar, R.V., Nataraju, A., Mohamed, R., Shivaprasad, H.V., Vishwanath, B.S., Anti-inflammatory activity of oleanolic acid by inhibition of secretory phospholipase A2 (2009) Planta Med, 75, pp. 211-215Magalhães, A., Santos, G.B., Verdam, M.C., Fraporti, L., Malheiro, A., Lima, E.S., Dos-Santos, M.C., Inhibition of the inflammatory and coagulant action of Bothrops atrox venom by the plant species Marsypianthes chamaedrys (2011) J Ethnopharmacol, 134, pp. 82-88Sampaio, S.C., Sousa-e-Silva, M.C.C., Borelli, P., Curi, R., Cury, Y., Crotalus durissus terrificus snake venom regulates macrophage metabolism and function (2001) J Leukoc Biol, 70, pp. 551-558(2002) Portuguese Pharmacopoeia, , Lisboa: Infarmed Editors, Portugal, Portuguese Pharmacopoeia CommitteeHarborne, J.B., (1998) Phytochemical Methods: A Guide to Modern Techniques of Plants Analysis, , London: Chapman & HallCintra Francischinelli, M., Silva, M.G., Andréo Filho, N., Cintra, A.C.O., Leite, G.B., Cruz Höfling, M.A.D.A., Rodrigues Simioni, L., Oshima Franco, Y., Effects of commonly used solubilizing agents on a model nerve-muscle synapse (2008) Lat Am J Pharm, 27, pp. 721-726Siles Villaroel, M., Rolim Rosa, R., Zelante, F., Furlanetto, R.S., Padronização da avaliação da potência de antivenenos botrópicos, em camundongos (1978) Mem Inst Butantan, 42-43, pp. 325-336Bülbring, E., Observation on the isolated phrenic nerve diaphragm preparation of the rat (1946) Br J Pharmacol, 1, pp. 38-61Ferraz, M.C., Parrilha, L.A.C., Moraes, M.S.D., Amaral Filho, J., Cogo, J.C., Dos Santos, M.G., Franco, L.M., Oshima Franco, Y., The effect of lupane triterpenoids (Dipteryx alata Vogel) in the in vitro neuromuscular blockade and myotoxicity of two snake venoms (2012) Curr Org Chem, 16, pp. 2717-2723Puebla, P., Oshima-Franco, Y., Franco, L.M., Santos, M.G., Silva, R.V., Rubem-Mauro, L., Feliciano, A.S., Chemical constituents of the bark of Dipteryx alata Vogel, an active species against Bothrops jararacussu venom (2010) Molecules, 15, pp. 8193-8204Gottlieb, O., Kaplan, M.A., Das plantas medicinais aos fármacos naturais (1993) Cienc Hoje, 15, pp. 51-54Souza Brito, A.R.M., How to study the pharmacology of medicinal plants in underdeveloped countries (1996) J Ethnopharmacol, 54, pp. 131-138Martini, N.D., Katerere, D.R., Eloff, J.N., Biological activity of five antibacterial flavonoids from Combretum erythrophyllum (Combretaceae) (2004) J Ethnopharmacol, 93, pp. 207-212Rates, S.M.K., Plants as source of drugs (2001) Toxicon, 39, pp. 603-613Soares, A.M., Ticli, F.K., Marcussi, S., Lourenço, M.V., Januário, A.H., Sampaio, S.V., Giglio, J.R., Pereira, O.S., Medicinal plants with inhibitory properties against snake venoms (2005) Curr Med Chem, 12, pp. 2625-2641Mors, W.B., Do Nascimento, M.C., Pereira, B.M.R., Pereira, N.A., Plant natural products active against snake bite - the molecular approach (2000) Phytochemistry, 55, pp. 627-642Barbosa, A.M., Villaverde, A.B., Guimarães Souza, L., Ribeiro, W., Cogo, J.C., Zamuner, S.R., Effect of low-level laser therapy in the inflammatory response induced by Bothrops jararacussu snake venom (2008) Toxicon, 1 (51), pp. 1236-1244Oshima-Franco, Y., Hyslop, S., Cintra, A.C., Giglio, J.R., da Cruz-Höfling, M.A., Rodrigues-Simioni, L., Neutralizing capacity of commercial bothropic antivenom against Bothrops jararacussu venom and bothropstoxin-I. (2000) Muscle Nerve, 23, pp. 1832-1839Oshima Franco, Y., Rosa, L.J.R., Silva, G.A.A., Amaral Filho, J., Silva, M.G., Lopes, P.S., Cogo, J.C., Da Cruz Höfling, M.A., Antibothropic action of Camellia sinensis extract against the neuromuscular blockade by Bothrops jararacussu snake venom and its mais toxin, bothropstoxin-I (2012) Pharmacology, pp. 469-489. , Croatia: Intech, Gallelli LMilani Júnior, R., Jorge, M.T., de Campos, F.P., Martins, F.P., Bousso, A., Cardoso, J.L., Ribeiro, L.A., Warrell, D.A., Snake bites by the jararacuçu (Bothrops jararacussu): clinic pathological studies of 29 proven cases in São Paulo State (1997) Brazil. QJM, 90, pp. 323-334Petricevich, V.L., Teixeira, C.F.P., Tambourghi, D.V., Gutiérrez, J.M., Increments in serum cytokine and nitric oxide levels in mice injected with Bothrops asper and Bothrops jararaca snake venom (2000) Toxicon, 38, pp. 1253-1266(2007) Rabies and envenomings: a neglected public health issue, , http://www.who.int/bloodproducts/animal_sera/Rabies.pdf, Available from: WHO - World Health OrganizationMalomouzh, A.I., Mukhtarov, M.R., Nikolsky, E.E., Vyskocil, F., Lieberman, E.M., Urazaev, A.K., Glutamate regulation of non-quantal release of acetylcholine in the rat neuromuscular junction (2003) J Neurochem, 85, pp. 206-213Rubem Mauro, L., Rocha, D.S., Barcelos, C.C., Varca, G.H., Andréo Filho, N., Barberato Filho, S., Oshima Franco, Y., Phenobarbital pharmacological findings on the nerve-muscle basis (2009) Lat Am J Pharm, 28, pp. 211-218. , Vila MMDCCintra-Francischinelli, M., Silva, M.G., Andréo-Filho, N., Gerenutti, M., Cintra, A.C.O., Giglio, J.R., Leite, G.B., Oshima-Franco, Y., Antibothropic action of Casearia sylvestris Sw. (Flacourtiaceae) extracts (2008) Phytother Res, 22, pp. 784-790Camargo, T.M., Nazato, V.S., Silva, M.G., Cogo, J.C., Groppo, F.C., Oshima-Franco, Y., Bothrops jararacussu venom-induced neuromuscular blockade inhibited by Casearia gossypiosperma Briquet hydroalcoholic extract (2010) J Venom Anim Toxins incl Trop Dis, 16, pp. 432-441Nazato, V.S., Rubem Mauro, L., Vieira, N.A.G., Rocha Junior, D., Dos, S., Silva, M.G., Lopes, P.S., Oshima Franco, Y., vitro antiophidian properties of Dipteryx alata Vogel bark extracts (2010) Molecules, 15, pp. 5956-5970Melo, R.F., Farrapo, N.M., Rocha Junior, D.S., Silva, M.G., Cogo, J.C., Dal Belo, C.A., Rodrigues Simioni, L., Oshima Franco, Y., Antiophidian mechanisms of medicinal plants (2009) Flavonoids: Biosynthesis, Biological Effects and Dietary Sources, pp. 249-262. , New York: Nova Science Publishers, Keller RBFarrapo, N.M., Silva, G.A.A., Costa, K.N., Silva, M.G., Cogo, J.C., Dal Belo, C.A., Dos Santos, M.G., Oshima Franco, Y., Inhibition of Bothrops jararacussu venom activities by Plathymenia reticulata Benth extracts (2011) J Venom Res, 2, pp. 52-58Cos, P., Vlietinc, A.J., Berghe, D.V., Maes, L., Anti-infective potential of natural products: how to develop a stronger in vitro 'proof-of-concept' (2006) J Ethnopharmacol, 103, pp. 290-302Mishra, U.S., Mishra, A., Kumari, R., Murthy, P.N., Naik, B.S., Antibacterial activity of ethanol extract of Andrographis paniculata (2009) Indian J Pharm Sci, 71, pp. 436-438Premendran, S.J., Salwe, K.J., Pathak, S., Brahmane, R., Manimekalai, K., Anti-cobra venom activity of plant Andrographis paniculata and its comparison with polyvalent anti-snake venom (2011) J Nat Sci Biol Med, 2, pp. 198-204Mahadeswaraswamy, Y.H., Nagaraju, S., Girish, K.S., Kemparaju, K., Local tissue destruction and procoagulation properties of Echis carinatus venom: inhibition by Vitis vinifera seed methanol extract (2008) Phytother Res, 22, pp. 963-969Oliveira, D.A., Salvador, A.A., Smânia, A., Smânia, E.F., Maraschin, M., Ferreira, S.R., Antimicrobial activity and composition profile of grape (Vitis vinifera) pomace extracts obtained by supercritical fluids (2013) J Biotechnol, 164, pp. 423-43

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