RESUMO
Studies on the interaction of dicholesteroyl diselenide (DCDS) and diphenyl diselenide (DPDS) with hepatic delta-aminolevulinic acid dehydratase (ALA-D) and different isoforms of lactate dehydrogenase (LDH) from different tissues were investigated. In addition, their antioxidant effects were tested in vitro by measuring the ability of the compounds to inhibit the formation of hepatic thiobarbituric acid reactive species (TBARS) induced by both iron (II) and sodium nitroprusside (SNP). The results show that while DPDS markedly inhibited the formation of TBARS induced by both iron (II) and SNP, DCDS did not. Also, the activities of hepatic delta-aminolevulinic acid dehydratase (ALA-D) and different isoforms of lactate dehydrogenase (LDH) were significantly inhibited by both DPDS and DCDS. Moreover, we further observed that the in vitro inhibition of different isoforms of lactate dehydrogenase by DCDS and DPDS likely involves the modification of the groups at the NAD+ binding site of the enzyme. Since organoselenides interacts with thiol groups on proteins, we conclude that the inhibition of different isoforms of lactate dehydrogenase by DPDS and DCDS possibly involves the modification of the thiol groups at the NAD+ binding site of the enzyme.
Assuntos
Antioxidantes/toxicidade , Derivados de Benzeno/toxicidade , Colesterol/análogos & derivados , Inibidores Enzimáticos/toxicidade , L-Lactato Desidrogenase/antagonistas & inibidores , Fígado/efeitos dos fármacos , Compostos Organosselênicos/toxicidade , Sintase do Porfobilinogênio/antagonistas & inibidores , Animais , Colesterol/toxicidade , Relação Dose-Resposta a Droga , Interações Medicamentosas , Coração/efeitos dos fármacos , Isoenzimas , L-Lactato Desidrogenase/metabolismo , Fígado/enzimologia , Masculino , Miocárdio/enzimologia , Nitroprussiato/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Sintase do Porfobilinogênio/metabolismo , Ratos , Ratos Wistar , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismoRESUMO
This paper evaluates the ability of organoselenium compounds [ebselen, selenocystine N-ethyl-carbamate (SeCis), bis-4-isopropyl-2-oxazolinyl phenyl diselenide (AASe)] to prevent HgCl(2) toxicity. Rats were injected with HgCl(2) (0 or 17 micromol/kg, sc) 6 h after organoselenium compounds had been injected (0 or 50 micromol/kg, sc). In vivo, HgCl(2) inhibited renal ALA-D activity ( approximately 48%), increased TBARS level in kidney ( approximately 52%) and reduced the hepatic content of non-protein thiol groups ( approximately 40%), but organoselenium compounds did not prevent such effects. SeCis, per se, increased renal TBARS level ( approximately 42%), while AASe increased hepatic content of ascorbic acid ( approximately 38%). In vitro, renal and hepatic ALA-D activity was inhibited by HgCl(2) (>or=25 microM), ebselen (>or=12 microM) and SeCis (>or=4 microM). HgCl(2) (400 microM) significantly increased TBARS production in renal and hepatic tissue preparations in vitro, and this effect was completely or partially prevented by organoselenium compounds. Ebselen exhibited thiol peroxidase activity in our assay conditions, while SeCis exhibited thiol-oxidizing properties regardless of the presence of peroxide. AASe had no effect on thiol oxidation. Results suggest that organoselenium compounds could not prevent mercury toxicity in vivo. The protective effect of these compounds against mercury-induced increase of TBARS production in vitro is probably related to an antioxidant action rather than to mercury binding.
Assuntos
Antioxidantes/farmacologia , Rim/enzimologia , Fígado/enzimologia , Mercúrio/toxicidade , Compostos Organosselênicos/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Sintase do Porfobilinogênio/metabolismo , Animais , Ácido Ascórbico/sangue , Azóis/farmacologia , Glutationa/metabolismo , Isoindóis , Rim/efeitos dos fármacos , Fígado/efeitos dos fármacos , Masculino , Peroxidases/metabolismo , Ratos , Ratos Wistar , Compostos de Sulfidrila/metabolismo , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismoRESUMO
This study investigated the hemolytic and genotoxic effect of different organoselenium and organotellurium compounds in human blood cells, as simple tests for screening the toxicity of organochalcogenides. For osmotic fragility (OF) test, samples of total blood were incubated with the organochalcogens at 4, 8, 50, 75 and 100 microM or vehicle (DMSO) for 90 min at 37 degrees C. The EC(50) values for hemolysis were significantly increased in erythrocytes exposed to diphenyl selenide (II), diphenyl diselenide (III), diphenyl telluride (IV), diphenyl ditelluride (V), (S)-2-amino-1-diselenide-3-methylpropanyl (IX), butyl(styryl)telluride (XIII) and 2-(butyltellurium)furan (XIV) at higher concentrations tested. The exposure of erythrocytes to organochalcogens diphenyl diselenide (II) and butyl(styryl)telluride (XIII), which had greater hemolytic effect, did not modify catalase activity, reactive oxygen species (ROS) production and -SH content. On the other hand, Na(+)/K(+) ATPase activity of erythrocyte ghosts was significantly inhibited by the compounds diphenyl diselenide (II) and butyl(styryl)telluride (XIII) (P<0.05) in a concentration-dependent manner. The inhibition of Na(+)/K(+) ATPase activity was completely reversed by dithiothreitol (DTT); indicating reaction of these organochalcogens with thiol groups of the enzyme. The thiol oxidase activity of the compounds II and XIII was supported by the fact that the rate of DTT oxidation was increased significantly by both chalcogens. In the higher concentrations, the compounds (II) and (XIII) were strongly genotoxic and cytotoxic to human leukocytes cells, as verified by the DNA damage and cell viability evaluation. Our results suggest that at relatively high concentration organochalcogenides exhibit hemolytic and genotoxic action in human blood cells, which are probably linked to their thiol oxidase activity and preferential interaction with sulfhydryl groups critical to enzymes.