In vitro and in vivo anti-inflammatory and antiophidic effects of the extract and fraction of Eugenia uniflora

Eugenia uniflora Linn, popularly known as ‘pitanga’, is a native plant endemic to Brazil that belongs to the Myrtaceae family. Its traditional use (leaves infusion) has been reported for the treatment of different diseases, including hypertension, inflammation, and as a diuretic agent. Considering the snakebite problem and the rich molecule repertoire of this herbal species, studies that evaluate its antiophidic potential are relevant for a broad social impact. This approach aims to evaluate the anti-inflammatory and antiophidic potential in vitro and in vivo of the extract (aqueous) and a fraction (ethyl acetate) of E. uniflora leaves against Bothrops leucurus and Bothrops brazili venoms. Extract and fraction from E. uniflora leaves were obtained by turbo-extraction and partitioning. The cytotoxicity was assayed on normal cell lines (Vero E6 and 3T3) using the 3-methyl-[4-5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide method. The anti-inflammatory activity of the aqueous extract was analyzed in vivo in the zymosan-induced air pouch model, and the leukocytes migration and other molecular inflammatory mediators quantified (myeloperoxidase, total protein, pro-inflammatory cytokine, malondialdehyde, and glutathione). In vitro, the antiophidic effect was evaluated by the ability of the E. uniflora extract and fraction to inhibit the enzymatic action (proteolytic, phospholipase A2, and hyaluronidase) of B. leucurus and B. brazili venoms. In addition, the antiophidic action in vivo was investigated after treatment with E. uniflora extract and fraction (50, 100, and 200 mg/kg) in the B. leucurus venom-induced paw edema with an evaluation of the antiedematogenic effect and quantification of myeloperoxidase (MPO) and pro-inflammatory cytokine levels. The E. uniflora leaves extract (7.8–125 mg/mL) revealed no toxicity in cell culture, but reduced MTT by 47% at the highest concentration (250 mg/mL) in Vero E6 cells. In contrast, the E. uniflora fraction (7.8–250 mg/mL) showed no cytotoxicity for both cell lines. In the air pouch model, E. uniflora leaves extract demonstrated anti-inflammatory activity, reducing cell migration, MPO activity, protein, malondialdehyde, and proinflammatory cytokines, and increased glutathione levels. Evaluating the antiophidic action in vitro, E. uniflora extract and fraction inhibited the proteolytic, phospholipase, and hyaluronidase effects of B. leucurus and B. brazili venoms at low concentrations. In addition, the extract an