Pérez GY, Oyarzábal YA, Ravelo CY, Mas FR, Jiménez DS, Molina CV

Language: Spanish
References: 26
Page: 259-266
PDF size: 54.89 Kb.

ABSTRACT

Introduction: D-002, a mixture of six high molecular weight primary aliphatic alcohols purified from beeswax, has been shown to produce anti-inflammatory effects with no secondary gastrotoxicity in experimental models. Oral treatment with D-002 was effective for lowering the concentrations of B4 leukotriene (LTB4) in pleural exudates of rats with carragenin-induced pleurisy, suggesting that it could inhibit 5-lipooxigenase (5-LOX) enzyme activity. The mechanisms of the antiinflammatory action of D-002, however, had not been explored yet.
Objective: to evaluate the effects of D-002 on 5-LOX enzyme activity in vitro by using the cytosolic preparations from rat liver homogenates.
Methods: testing conditions were as follows: cytosolic fraction (50 μg of protein) dissolved in 0.2 mol/L borate buffer solution (pH 9) and linoleic acid (7.8-250 mmol/L) as substrate. Parallel samples were incubated with Tween-20/H2O (2 %) only (vehicle, control samples), D-002 (0.9-1 000 μg/mL) or Lyprinol (500 μg/mL) (reference substance). The enzyme activity, evaluated through the formation of conjugated dienes, was assessed by the absorbance changes at 234 nm (5-lox) measured in a UV-visible spectrophotometer.
Results: the in vitro addition of D-002 produced a significant, dose-dependent (r=0.980; p< 0.001) (IC50= 95.34 μg/mL) and uncompetitive inhibition of 5-LOX activity, whose maximal inhibition (70 %) was achieved with 500 μg/mL).
Conclusions: this study demonstrates that D-002 effectively inhibits 5-LOX enzymatic activity, an effect that may partially explain the anti-inflammatory effects of D-002 in experimental models in vivo.

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