RESUMEN
Aldose reductase (AKR1B1), the first enzyme in the polyol pathway, is likely involved in the onset of diabetic complications. Differential inhibition of AKR1B1 has been proposed to counteract the damaging effects linked to the activity of the enzyme while preserving its detoxifying ability. Here, we show that epigallocatechin gallate (EGCG), one of the most representative catechins present in green tea, acts as a differential inhibitor of human recombinant AKR1B1. A kinetic analysis of EGCG, and of its components, gallic acid (GA) and epigallocatechin (EGC) as inhibitors of the reduction of L-idose, 4-hydroxy2,3-nonenal (HNE), and 3-glutathionyl l-4-dihydroxynonanal (GSHNE) revealed for the compounds a different model of inhibition toward the different substrates. While EGCG preferentially inhibited L-idose and GSHNE reduction with respect to HNE, gallic acid, which was still active in inhibiting the reduction of the sugar, was less active in inhibiting HNE and GSHNE reduction. EGC was found to be less efficient as an inhibitor of AKR1B1 and devoid of any differential inhibitory action. A computational study defined different interactive modes for the three substrates on the AKR1B1 active site and suggested a rationale for the observed differential inhibition. A chromatographic fractionation of an alcoholic green tea extract revealed that, besides EGCG and GA, other components may exhibit the differential inhibition of AKR1B1.
Asunto(s)
Aldehído Reductasa/metabolismo , Catequina/análogos & derivados , Inhibidores Enzimáticos/farmacología , Té/química , Aldehído Reductasa/química , Dominio Catalítico/efectos de los fármacos , Catequina/química , Catequina/farmacología , Inhibidores Enzimáticos/química , Ácido Gálico/química , Ácido Gálico/farmacología , Glutatión/análogos & derivados , Glutatión/metabolismo , Hexosas/metabolismo , Humanos , Modelos Moleculares , Simulación del Acoplamiento Molecular , Simulación de Dinámica Molecular , Extractos Vegetales/química , Extractos Vegetales/farmacologíaRESUMEN
The hyperactivity of aldose reductase (AR) on glucose in diabetic conditions or on glutathionyl-hydroxynonenal in oxidative stress conditions, the source of cell damage and inflammation, appear to be balanced by the detoxifying action exerted by the enzyme. This detoxification acts on cytotoxic hydrophobic aldehydes deriving from membrane peroxidative processes. This may contribute to the failure in drug development for humans to favorably intervene in diabetic complications and inflammation, despite the specificity and high efficiency of several available aldose reductase inhibitors. This paper presents additional features to a previously proposed approach, on inhibiting the enzyme through molecules able to preferentially inhibit the enzyme depending on the substrate the enzyme is working on. These differential inhibitors (ARDIs) should act on glucose reduction catalyzed by AR without little or no effect on the reduction of alkenals or alkanals. The reasons why AR may be an eligible enzyme for differential inhibition are considered. These mainly refer to the evidence that, although AR is an unspecific enzyme that recognizes different substrates such as aldoses and hydrophobic aldehydes, it nevertheless displays a certain degree of specificity among substrates of the same class. After screening on edible vegetables, indications of the presence of molecules potentially acting as ARDIs are reported.
Asunto(s)
Aldehído Reductasa/metabolismo , Inhibidores Enzimáticos/metabolismo , Verduras/química , Aldehído Reductasa/antagonistas & inhibidores , Aldehído Reductasa/genética , Inhibidores Enzimáticos/química , Glucosa/metabolismo , Humanos , Phaseolus/química , Phaseolus/metabolismo , Extractos Vegetales/química , Extractos Vegetales/metabolismo , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/química , Proteínas Recombinantes/aislamiento & purificación , Especificidad por Sustrato , Verduras/metabolismoRESUMEN
Arabinogalactan is a naturally-occurring, densely branched, polysaccharide mainly made-up of galactose and arabinose with variable amounts of uronic acids, which received attention for several industrial and biomedical applications. The ability of Western Larch arabinogalactan to interact with mucins was assessed by both classical gel filtration chromatography and frontal chromatography on Sephacryl S300 resin. The shift of arabinogalactan elution volume in classical gel filtration chromatography induced by both bovine submaxillary mucin and porcine gastric mucin resulted useful for revealing the occurrence of an interaction between arabinogalactan and mucins. A frontal gel chromatography, in which arabinogalactan is used as eluent, enabled a dissociation constant of 5×10(-6)M to be measured for the arabinogalactan-bovine submaxillary mucin complex, with approximately 50 equivalents of arabinogalactan bound per mucin mole. The mucoadhesivity of arabinogalactan may be a relevant feature for its biomedical and industrial applications.