Effects of leaf and seed alcoholic extracts from of annona crassiflora mart. (annonaceae) in mice

Annona crassiflora Mart. is a species of the cerrado biome in the family Annonaceae. It is a vegetable with therapeutic potential. However, it should be emphasized that its toxicity when consumed in large doses remains unknown, and clinical studies are strongly suggested for this purpose. The objective of this study was to verify the toxicological effect of leaf and seed extracts of Annona crassiflora Mart. (Annonaceae) in mice. The plant was collected in were collected in cerrado areas in Montes Claros, Minas Gerais, Brazil. Healthy leaves and seeds were selected and dried in a forced air circulating dryer at 40ºC for 72 h. Ethanolic extract (EE) was obtained by maceration of the dried leaves in absolute ethanol. The extracts were filtered and subsequently dehydrated at 40ºC for 48 h. The chromatographic profile was performed using an HPLC system. To evaluate the toxicological effects, 18 Swiss mice, were divided into three groups, composed of three males and three females each as follows: 22 μL PBS (C), 22 μL leaf extract (LE), 22 μL seed extract (SE) that were administered by means of gavage at cumulative doses for four days at the following concentrations: day one, 12.5mg/kg; day two, 25.0mg/kg; day three 50mg/kg and the fourth day, 100mg/kg. To assess the toxicological effects, physical aspects, behavior, and body weight of animals were monitored daily. On the fifth day the mice were euthanized and peripheral blood and target tissues were collected. ALT plasma level was measured. Liver, spleen, heart, and kidney samples were collected, formalin-fisxed, paraffin-embedded, and submited to histopathological analysis. Statistical analysis was performed using Student t tests and ANOVA. The Bonferroni test was applied as post hoc. Valeus of p < 0.05 were considered as statistically significant. Our findings showed that A. crassiflora Mart. extract is rich in flavonoids and flavones. No clinical signs or symptoms that had suggested toxicity were observed in all animals. However LE and SE animals showed decreased body weight, increased size and wight for liver and spleen organs. Moreover, ALT levels were increased in experimental animals. Histological analysis showed morphological findings of discrete hyperemia in liver and spleen tissues of animals treated with SE. In conclusion, the present study evidenced potential toxic effects of A.crassiflora Mart. leaf and seed extract notably in liver and spleen organs. More prospective studies are needed to better understand the cytotoxic effects of A. crassiflora.