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Evid Based Complement Alternat Med. 2015;2015:939523. doi: 10.1155/2015/939523. Epub 2015 Nov 8.

The Triterpenoid Betulin Protects against the Neuromuscular Effects of Bothrops jararacussu Snake Venom In Vivo.

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Post-Graduate Program in Pharmaceutical Sciences and Pharmacy Course, University of Sorocaba (UNISO), Rodovia Raposo Tavares, Km 92,5, 18023-000 Sorocaba, SP, Brazil.
Serpentarium of the Center for Nature Studies and Institute for Research and Development (IP&D), Vale do Paraíba University (UNIVAP), Avenida Shishima Hifumi 291, 12244-000 São José dos Campos, SP, Brazil.
Post-Graduate Program in Environmental Sciences, Tocantins Federal University, Avenida NS15, ALC NO14, 109 Norte, 77001-090 Palmas, TO, Brazil.
Pharmacy Course, Methodist University of Piracicaba (UNIMEP), Rodovia do Açúcar, Km 156, 13423-170 Piracicaba, SP, Brazil.
Department of Pharmaceutical Chemistry, Salamanca University, Campus "Miguel de Unamuno", 37007 Salamanca, Spain.
Pharmacy Course, Federal University of São Paulo (UNIFESP), Rua Prof. Artur Riedel 275, 09972-270 Diadema, SP, Brazil.
Department of Pharmacy, Faculty of Pharmaceutical Sciences, University of São Paulo (USP), Avenida Prof. Lineu Prestes 580, 05434-070 São Paulo, SP, Brazil.
Laboratory of Herpetology, Instituto Butantan, Avenida Vital Brasil 1500, 05503-900 São Paulo, SP, Brazil.
Department of Pharmacology, Faculty of Medical Sciences, State University of Campinas (UNICAMP), Rua Tessália Vieira de Camargo 126, Cidade Universitária Zeferino Vaz, 13083-870 Campinas, SP, Brazil.


We confirmed the ability of the triterpenoid betulin to protect against neurotoxicity caused by Bothrops jararacussu snake venom in vitro in mouse isolated phrenic nerve-diaphragm (PND) preparations and examined its capability of in vivo protection using the rat external popliteal/sciatic nerve-tibialis anterior (EPSTA) preparation. Venom caused complete, irreversible blockade in PND (40 μg/mL), but only partial blockade (~30%) in EPSTA (3.6 mg/kg, i.m.) after 120 min. In PND, preincubation of venom with commercial bothropic antivenom (CBA) attenuated the venom-induced blockade, and, in EPSTA, CBA given i.v. 15 min after venom also attenuated the blockade (by ~70% in both preparations). Preincubation of venom with betulin (200 μg/mL) markedly attenuated the venom-induced blockade in PND; similarly, a single dose of betulin (20 mg, i.p., 15 min after venom) virtually abolished the venom-induced decrease in contractility. Plasma creatine kinase activity was significantly elevated 120 min after venom injection in the EPSTA but was attenuated by CBA and betulin. These results indicate that betulin given i.p. has a similar efficacy as CBA given i.v. in attenuating the neuromuscular effects of B. jararacussu venom in vivo and could be a useful complementary measure to antivenom therapy for treating snakebite.

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