Bárbara Ricciardi Verrastro1, Ana Maria Torres2, Gabriela Ricciardi3, Pamela Teibler4, Silvana Maruñak5, Chiara Barnaba6, Roberto Larcher7, Giorgio Nicolini8, Eduardo Dellacassa9. 1. Facultad de Ciencias Exactas y Naturales y Agrimensura, Universidad Nacional del Nordeste, Av. Libertad 5470, 3400 Corrientes, Argentina. Electronic address: barb_ricciardi@hotmail.com. 2. Facultad de Ciencias Exactas y Naturales y Agrimensura, Universidad Nacional del Nordeste, Av. Libertad 5470, 3400 Corrientes, Argentina. Electronic address: amtorres@exa.unne.edu.ar. 3. Facultad de Ciencias Exactas y Naturales y Agrimensura, Universidad Nacional del Nordeste, Av. Libertad 5470, 3400 Corrientes, Argentina. Electronic address: gricciardi@exa.unne.edu.ar. 4. Facultad de Veterinaria, Universidad Nacional del Nordeste, Sargento Cabral 2139, 3400 Corrientes, Argentina. Electronic address: serviciofarmacia@vet.unne.ed.ar. 5. Facultad de Veterinaria, Universidad Nacional del Nordeste, Sargento Cabral 2139, 3400 Corrientes, Argentina. Electronic address: patmed@vet.unne.edu.ar. 6. Facultad de Química, Universidad de la República, Av General Flores 2124, 11800 Montevideo, Uruguay. Electronic address: chiara.barnaba@fmach.it. 7. Facultad de Química, Universidad de la República, Av General Flores 2124, 11800 Montevideo, Uruguay. Electronic address: roberto.larcher@fmach.it. 8. Facultad de Química, Universidad de la República, Av General Flores 2124, 11800 Montevideo, Uruguay. Electronic address: giorgio.nicolini@fmach.it. 9. Centro Trasferimento Tecnologico, Fondazione Edmund Mach di San Michele all'Adige, Via E. Mach 1, 38010-S Michele all'Adige, TN, Italy. Electronic address: edellac@fq.edu.uy.
Abstract
ETHNOPHARMACOLOGICAL RELEVANCE: Ophidian accidents are a serious public health problem in Argentina; the Bothrops species is responsible for 97% of these accidents, and in particular, B. diporus is responsible for 80% of them. In the northeast of the country (Corrientes Provinces), Cissampelos pareira L. (Menispermaceae) is commonly used against the venom of B. diporus; its use is described in almost all ethnobotanical literature from countries where the plant grows. AIM OF THE STUDY: In this study, the in vitro and in vivo antivenom activities of C. pareira extracts were evaluated against B. diporus venom, with a particular focus on the local effects associated with envenoming. The seasonal influence on the chemical composition of the active extracts was also studied, in order determine the associated range of variability and its influence on the antivenom activity. MATERIALS AND METHODS: This research was conducted using aerial parts (leaves, flowers, tender stems) and roots of Cissampelos pareira collected from two different phytogeographic regions of Corrientes (Argentina); Paso de la Patria and Lomas de Vallejos. In addition, to perform a seasonal analysis and to evaluate the metabolic stability, material was collected at three different growth stages. In vivo and in vitro anti-snake venom activities were tested, and a bio-guided chromatographic separation was performed in order to determine the active chemicals involved. The fractions obtained were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and the chemical profile of the most active constituent was analyzed by ultra high-performance liquid chromatography coupled to quadrupole/high-resolution mass spectrometry (Q-Orbitrap). (UHPLC-MS). RESULTS: The alcoholic extract was found to be the most active The bio-guided fractionation allowed selection one fraction to be analyzed by UHPLC-MS in order to identify the components responsible for the activities found; this identified five possible flavonoids. CONCLUSIONS: Our studies of the activity of C. pareira against the venom of B. diporus have confirmed that this species possesses inhibitory effects in both in vitro and in vivo models. Moreover, the present data demonstrate that certain flavonoids may mitigate some of the venom-induced local tissue damage.
ETHNOPHARMACOLOGICAL RELEVANCE: Ophidian accidents are a serious public health problem in Argentina; the Bothrops species is responsible for 97% of these accidents, and in particular, B. diporus is responsible for 80% of them. In the northeast of the country (Corrientes Provinces), Cissampelos pareira L. (Menispermaceae) is commonly used against the venom of B. diporus; its use is described in almost all ethnobotanical literature from countries where the plant grows. AIM OF THE STUDY: In this study, the in vitro and in vivo antivenom activities of C. pareira extracts were evaluated against B. diporus venom, with a particular focus on the local effects associated with envenoming. The seasonal influence on the chemical composition of the active extracts was also studied, in order determine the associated range of variability and its influence on the antivenom activity. MATERIALS AND METHODS: This research was conducted using aerial parts (leaves, flowers, tender stems) and roots of Cissampelos pareira collected from two different phytogeographic regions of Corrientes (Argentina); Paso de la Patria and Lomas de Vallejos. In addition, to perform a seasonal analysis and to evaluate the metabolic stability, material was collected at three different growth stages. In vivo and in vitro anti-snake venom activities were tested, and a bio-guided chromatographic separation was performed in order to determine the active chemicals involved. The fractions obtained were analyzed by sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) and the chemical profile of the most active constituent was analyzed by ultra high-performance liquid chromatography coupled to quadrupole/high-resolution mass spectrometry (Q-Orbitrap). (UHPLC-MS). RESULTS: The alcoholic extract was found to be the most active The bio-guided fractionation allowed selection one fraction to be analyzed by UHPLC-MS in order to identify the components responsible for the activities found; this identified five possible flavonoids. CONCLUSIONS: Our studies of the activity of C. pareira against the venom of B. diporus have confirmed that this species possesses inhibitory effects in both in vitro and in vivo models. Moreover, the present data demonstrate that certain flavonoids may mitigate some of the venom-induced local tissue damage.