Evaluation of the Analgesic and Anti-Inflammatory Effects of a Brazilian Green Propolis

Abstract

Phamacological activities of a standard ethanol extract G1 from Brazilian green propolis, typified as BRP1, was evaluated in mouse models of pain and inflammation. Intraperitoneal injection (i. p.) of G1 inhibited acetic acid-induced abdominal constrictions with an ID₅₀ = 0.75 ± 0.05 mg/kg, and in the formalin test the ID₅₀ values were 0.85 ± 0.07 mg/kg and 13.88 ± 1.12 mg/kg, respectively, for the neurogenic and inflammatory phases. The extract was ineffective when assessed in the hot-plate assay. In serotonin-induced paw edema, G1 led to a maximal inhibition (MI) of 51.6 % after 120 min when administered i. p. and of 36 % after 15 min by the oral route (o. r.). When the inflammatory agent was complete Freund’s adjuvant, inhibition of paw edema was also observed after administration of the extract by both routes. In the capsaicin-induced ear edema the ID₅₀ values were 1.09 ± 0.08 mg/kg (i. p.) and 10.00 ± 0.90 mg/kg (o. r.). In the acute carrageenan-induced inflammatory reaction induced by carrageenan, G1 reduced the number of neutrophils in the peritoneal cavity with IC₅₀ values of 0.72 ± 0.08 mg/kg and 4.17 ± 0.50 mg/kg, by i. p. or o. r. administration, with a preferential migration of polymorphonuclear neutrophils. In vitro, G1 decreased nitric oxide production in LPS-stimulated RAW 264.7 cells (IC₅₀ = 41.60 μg/mL), and also the luciferase activity in TNF-α-stimulated HEK 293 cells transfected with NF-κB-luciferase reporter gene driven by the nuclear factor κB (NF-κB) (IC₅₀ = 200 μg/mL). This extract, which at low concentrations induces anti-inflammatory and analgesic effects in mouse models, presents a high content of flavonoids, known to inhibit inducible NOS (iNOS) activity. These data taken together led us to reinforce the hypothesis in the literature that the anti-inflammatory effect of propolis may be a due to inhibition of iNOS gene expression, through interference with NF-κB sites in the iNOS promoter.

Abbreviations

CFA:complete Freund adjuvant

COX-2:inducible isoform of cyclooxygenase

Indo:Indomethacin

iNOS:inducible nitric oxide synthase

LPS:Lipopolysaccharide

MI:maximal inhibition

MTT:3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

NF-κB:nuclear factor κB

NO:nitric oxide

TMB:3,3′,5,5′-tetramethylbenzidine

Key words

Typified propolis - analgesic effect - anti-inflammatory effect - NF-κB factor - nitric oxide

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