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Efecto in vitro del D-002 sobre la actividad enzimática de la 5-lipoxigenasa (5-LOX)

Keywords: d-002, anti-inflammatory, 5-lox inhibition, ltb4, lyprinol.

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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 anti-inflammatory 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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