Avaliação do potencial anti-inflamatório e antioxidante do extrato seco padronizado (CLAE-DAD) de Amburana cearensis (cumaru) cultivada e constituintes químicos em neutrófilos humano

Abstract

Previous studies performed in the Center of Pharmaceutical and Cosmetical Studies (CEFAC) proved the anti-inflammatory and antioxidant potential of wild Amburana cearensis (cumaru). However, under the risk of extinction of the species due to its use in popular medicine and logging, studies have been conducted by our group, where the cultivated species showed chemical and biological similarity with the wild one. The objective of this present work was to evaluate the anti-inflammatory and antioxidant activity of standardized dry extract of cultivated Amburana cearensis (ESPACC) and chemical constituents, vanillic acid (AV) and coumarin (CM), aiming at its potential in the treatment of inflammatory diseases. The ESPACC was produced from the 7 months old cultivated plant and the marker content was determined by HPLC-DAD (AV: 2.55 mg/g and CM: 14.25 mg/g). The human neutrophils used were isolated from whole blood fraction donated by Hematology and Hemotherapy Center of Ceará (HEMOCE) (CAAE 45668515.4.0000.5054). The effects of ESPACC and AV (1, 10, 25, 50 and 100 μg / mL) over PMA induced neutrophil degranulation were measured by the release of the MPO enzyme which was reduced in up to 51.2% by ESPACC and 87.6% by AV, while indomethacin (36 μg/mL), the standard drug, reduced about 78,1%. The evaluation of the effects of ESPACC, CM and AV on the oxidative metabolism of neutrophils activated by PMA was measured by chemiluminescence assay (luminol and lucigenin probes) to quantify the reactive oxygen species (EROs). The cytotoxicity of the drugs was assessed by the MTT test. The addition of ESPACC at the highest concentration reduced the production of EROs luminol-marked by 62.7% and lucigenin by 15.4%, quercetin (50 μg / mL), standard antioxidant, reduced the amount of 71.3% and 19.73% respectively. The AV (100μg / mL) lowered the production of EROs marked with luminol in 63.8% and by lucigenin in 13.9%, comparing with quercetin that decreased in 63.3% and 12.9%, respectively. The CM reduced to a lesser degree (up to 24%) the production of luminolmarked EROs and did not influence the production of lucigenin-marked EROs. None of the concentrations investigated interfered significantly with neutrophil cell viability in relation to the control group. Thus, the results evidenced that the anti-inflammatory effect is related, at least in part, to the presence of CM and AV, suggesting that they have potential in the treatment of inflammatory diseases.