Toxicity and genotoxicity evaluation of Passiflora alata Curtis (Passifloraceae)
Graphical abstract
An Passiflora alata aerial parts extract (2.4% total flavonoids) induces DNA damage assessed by comet assay in mice peripheral blood leukocytes.
Introduction
The pharmacological knowledge of the genus Passiflora indicates its potential to the development of anxiolytic and sedative drugs (Dhawan et al., 2004). Passiflora incarnata is approved in Europe for treating nervous restlessness (Blumenthal et al., 2000) and also has been claimed to be useful in the treatment of obesity due to its anxiolytic properties (Moro and Basile, 2000). This specie did not show genotoxic effects in the in vitro assay with the Aspergillus nidulans as well as in the in vivo micronucleus induction test in mice (Parra et al., 2002). Nevertheless there is notice about genotoxicity of harman and harmine that are alkaloids present in the genus Passiflora (Boeira et al., 2001, Boeira et al., 2002).
Passiflora alata is the official species of Brazilian Pharmacopoeia and its aerial parts are used as medicinal plant by local population as well as constitutes several phytomedicines commercialized in Brazil as sedatives (ANVISA, 2008). Previous studies demonstrated anxiolytic and/or hypnotic activity for aqueous and hidroethanolic extracts of Passiflora alata (100–600 mg/kg i.p. and v.o.) in rodents (Oga et al., 1984, Petry et al., 2001, De Paris et al., 2002, Barbosa et al., 2008, Provensi et al., 2008). The aqueous leaves extracts of Passiflora alata (100–300 mg/kg, i.p.) showed a significant anti-inflammatory activity on carrageenan-induced pleurisy in mice (Vargas et al., 2007). Amaral et al. (2001) showed that Passiflora alata aqueous lyophilized extract (800 mg/kg/day, p.o.) did not present reproductive toxicity in female rats. As far as we know studies about genotoxicity of Passiflora alata extracts were not reported.
The most used genotoxicity tests in mammals are the comet and micronucleus. The former is a genotoxicity assay, which detects DNA strand breaks, alkali-labile sites and incomplete excision repair events in individual cells. It is a rapid and sensitive method for the detection of DNA damage induced in vitro or in vivo by various agents in a variety of cells (Tice et al., 2000, Collins, 2004, Moller, 2006). The later provides a simple and rapid indirect measure of induced structural and numerical chromosome aberrations and it is scientifically and regulatory accepted by supranational authorities like the Organization for Economic Cooperation and Development (OECD), International Conference on Harmonization (ICH) and European Union (EU) (Mavournin et al., 1990).
The aim of this study was to evaluate rodent acute and sub-acute toxicity and genotoxicity of a spray-dried extract of Passiflora alata leaves (PA) by using in vivo comet assay and micronucleus test in mice.
Section snippets
Plant material
The leaves of Passiflora alata were collected in the city of Canoas, State of Rio Grande do Sul (RS), Brazil (November 2002), and identified by Lilian Auler Mentz (Programa de Pós-Graduação em Botânica, UFRGS), voucher specimen ICN133727.
Preparation and characterization of extract
Passiflora alata leaves (560 g) were dried at 40 °C for 14 days. After drying, they were triturated and extracted with distilled water (plant:solvent, 1:10, w/v) under reflux for 1 h. This extract was filtered and then dried in Spray Dryer LMMSD 1.0 yielding 60 g
Characterization of extract and general toxicity
Flavonoids and saponins were detected as major constituents of the extract. Total flavonoid content was determined as 2.60% (w/v). Regarding the saponins, it was evidenced the presence of quadranguloside. Alkaloids were not detected.
No mice death occurrences were registered up to 4800 mg/kg p.o. at the first 24 h after treatment. Consequently it was not possible to determine the LD50 in mice. General toxicity signs were not observed in mice (PA 600–4800 mg/kg) as well as in rats after single and
Discussion
This work was carried out in an effort to evaluate the acute and sub-acute toxicity and genotoxicity of a spray-dried extract of Passiflora alata leaves (PA). Comet assay and micronucleus test were used to determine PA effects concerning DNA damage measured in peripheral blood cells and bone marrow of mice.
The dose range tested was chosen according to those cited for Passiflora alata extracts as effective in animal models of anxiety (Petry et al., 2001, De Paris et al., 2002, Barbosa et al.,
Acknowledgements
This work was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) and Departamento de Ciências da Saúde, URI, Erechim, Brazil. The authors are grateful to CAPES (Brazil) and PROPESQ-UFRGS (Brazil) for the fellowships to Raquel Fenner, Gustavo Provensi and Andresa H. Betti. We thank Carla Totti, Luciani Polidoro and Darieli Alves Moreira from Laboratório de Genética, Departamento de Ciências da Saúde URI, Erechim, Brazil, for technical assistance.
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