CONICET | Buscador de Institutos y Recursos Humanos

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. Materialsandmethods: Thisresearchwasconductedusingaerialparts(leaves, ﬂowers,tenderstems)androotsof 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: ThealcoholicextractwasfoundtobethemostactiveThebio-guidedfractionationallowedselectionone 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.