RESUMEN
Two chemotypes were examined in vitro with CYPs 3A4 and 2C19 by molecular docking, metabolic profiles, and intrinsic clearance deuterium isotope effects with specifically deuterated form to assess the potential for enhancement of pharmacokinetic parameters. The results show the complexity of deuteration as an approach for pharmacokinetic enhancement when CYP enzymes are involved in metabolic clearance. With CYP3A4 the rate limiting step was chemotype-dependent. With one chemotype no intrinsic clearance deuterium isotope effect was observed with any deuterated form, whereas with the other chemotype the rate limiting step was isotopically sensitive, and the magnitude of the intrinsic clearance isotope effect was dependent on the position(s) and extent of deuteration. Molecular docking and metabolic profiles aided in identifying sites for deuteration and predicted the possibility for metabolic switching. However, the potential for an isotope effect on the intrinsic clearance cannot be predicted and must be established by examining select deuterated versions of the chemotypes. The results show how in a deuteration strategy molecular docking, in-vitro metabolic profiles, and intrinsic clearance assessments with select deuterated versions of new chemical entities can be applied to determine the potential for pharmacokinetic enhancement in a discovery setting. They also help explain the substantial failures reported in the literature of deuterated versions of drugs to elicit a systemic enhancement on pharmacokinetic parameters.
Asunto(s)
Citocromo P-450 CYP2C19/química , Citocromo P-450 CYP3A/química , Deuterio/química , Farmacocinética , Citocromo P-450 CYP2C19/efectos de la radiación , Citocromo P-450 CYP3A/efectos de la radiación , Deuterio/farmacología , Hemo/química , Hemo/efectos de la radiación , Humanos , Inactivación Metabólica , Cinética , Microsomas/efectos de la radiación , Simulación del Acoplamiento Molecular , Oxidación-Reducción/efectos de la radiación , Especificidad por SustratoRESUMEN
Low-intensity light therapy (LILT) is showing promise in the treatment of a wide variety of medical conditions. Concurrently, our knowledge of LILT mechanisms continues to expand. We are now aware of LILT's potential to induce cellular effects through, for example, accelerated ATP production and the mitigation of oxidative stress. In clinical use, however, it is often difficult to predict patient response to LILT. It appears that cellular reduction/oxidation (redox) state may play a central role in determining sensitivity to LILT and may help explain variability in patient responsiveness. In LILT, conditions associated with elevated reactive oxygen species (ROS) production, e.g. diabetic hyperglycemia, demonstrate increased sensitivity to LILT. Consequently, assessment of tissue redox conditions in vivo may prove helpful in identifying responsive tissues. A noninvasive redox measure may be useful in advancing investigation in LILT and may one day be helpful in better identifying responsive patients. The detection of biophotons, the production of which is associated with cellular redox state and the generation of ROS, represents just such an opportunity. In this review, we will present the case for pursuing further investigation into the potential clinical partnership between biophoton detection and LILT.
Asunto(s)
Terapia por Luz de Baja Intensidad , Oxidación-Reducción/efectos de la radiación , Rayos Ultravioleta , Adenosina Trifosfato/metabolismo , Animales , Humanos , Peroxidación de Lípido , Microsomas/efectos de la radiación , Mitocondrias/efectos de la radiación , Estrés Oxidativo , Fotones , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Methyl paraben (MP) is often used as a preservative in foods, drugs, and cosmetics because of its high reliability in safety based on the rapid excretion and nonaccumulation following administration. Light irradiation sometimes produces unexpected activity from chemicals such as MP; furthermore, there is ample opportunity for MP to be exposed to sunlight. Here, we investigated whether MP shows DNA damage after sunlight irradiation. Two major photoproducts, p-hydroxybenzoic acid (PHBA) and 3-hydroxy methyl paraben (MP-3OH), were detected after sunlight irradiation to an aqueous MP solution. Both photoproducts were inactive in the in vitro DNA damage assay that measures oxidized guanine formed in calf thymus DNA in the presence of divalent copper ion, a known mediator of oxidative DNA damage. Simulated MP metabolism using dermal tissues after light irradiation produced these two photoproducts, which reacted with a microsomal fraction (S9) of the skin. A metabolite from MP-3OH, not PHBA, caused distinct DNA damage in the in vitro assay. This active metabolite was identified as protocatechuic acid, a hydrolyzed MP-3OH product. In addition, NADH, a cellular reductant, enhanced DNA damage by approximately five times. These results suggest that reactive oxygen species generated by the redox cycle via metal ion and catechol autoxidation are participating in oxidative DNA damage. This study reveals that MP might cause skin damage involving carcinogenesis through the combined activation of sunlight irradiation and skin esterases.
Asunto(s)
Daño del ADN , Esterasas/metabolismo , Parabenos/efectos de la radiación , Conservadores Farmacéuticos/efectos de la radiación , Animales , Bovinos , Cromatografía de Gases y Espectrometría de Masas , Hidroxibenzoatos/química , Masculino , Microsomas/metabolismo , Microsomas/efectos de la radiación , Oxidación-Reducción , Parabenos/química , Fotoquímica , Conservadores Farmacéuticos/química , Ratas , Ratas Sprague-Dawley , Especies Reactivas de Oxígeno/química , Piel/química , Piel/enzimología , Piel/efectos de la radiación , Rayos UltravioletaRESUMEN
Gallic acid (3,4,5-trihydroxybenzoic acid, GA) is a naturally occurring plant phenol. In vitro and in vivo studies have shown that this phytochemical protected DNA and membranes against ionizing radiation. Rat liver microsomes and plasmid pBR322 DNA were exposed to various doses of gamma radiation in presence and absence of GA. Exposure of the microsomes to gamma radiation resulted in the formation of peroxides of membrane lipids measured as thiobarbituric acid reactive substances and presence of GA during irradiation prevented the formation of lipid peroxidation. Gamma irradiation of plasmid DNA resulted in induction of strand breaks in DNA resulting in disappearance of the supercoiled (ccc) form. Presence of GA during irradiation protected the DNA from undergoing the strand breaks. In in vivo studies it was found that whole body exposure of mice to gamma radiation (4 Gy) increased the formation of lipid peroxides in various tissues and damage to cellular DNA (as measured by alkaline comet assay) in peripheral blood leucocytes. Administration of GA to mice prior to whole body radiation exposure reduced the peroxidation of lipids and the damage to the cellular DNA indicating in vivo radiation protection of membranes and DNA by GA.
Asunto(s)
ADN/efectos de la radiación , Ácido Gálico/farmacología , Membranas/efectos de la radiación , Animales , Ensayo Cometa , ADN/metabolismo , Relación Dosis-Respuesta a Droga , Flavonoides/antagonistas & inhibidores , Flavonoides/metabolismo , Ácido Gálico/metabolismo , Humanos , Dosificación Letal Mediana , Peroxidación de Lípido/efectos de la radiación , Peróxidos Lipídicos/metabolismo , Membranas/metabolismo , Ratones , Microsomas/metabolismo , Microsomas/efectos de la radiación , Fenoles/antagonistas & inhibidores , Fenoles/metabolismo , Polifenoles , Ratas , Especificidad de la Especie , Sustancias Reactivas al Ácido Tiobarbitúrico/metabolismoRESUMEN
The influence of irradiation and NSE on the microsomal lipid composition of the rat liver and heart was studied. It was shown, that radiation treatment in a dose of 2 Gy had not a significant affect on the heart phospholipid composition, whereas in the liver the amounts of the phosphatidylethnolamine and phosphatidylinositol were increased and the amounts of the phosphatidylcholine and sphyngomieline were decreased. NSE did not impact on these characteristics. The alterations of the plasmalogen/diacyl forms ratio of the PC and PE also took place only in the liver microsome. The analysis of the fatty acids esterified to phospholipids showed similar situation: the fatty acids of the heart microsome less were changed than those of the liver microsome undergo irradiation. The amount of arachidonic acid rise after the adding of NSE to rats. Investigation of the important component of biological membranes--cholesterol and its esters also showed that the liver tissue is more vulnerable to irradiation than the heart. NSE led to insignificant increase of the not esterified cholesterol in the liver. As a result of the present work we may conclude that the liver microsome is more vulnerable than the heart under X-ray radiation and NSE has protective effect in these conditions.
Asunto(s)
Etanolaminas/farmacología , Corazón , Microsomas Hepáticos , Microsomas , Fosfolípidos/metabolismo , Protectores contra Radiación/farmacología , Ácidos Esteáricos/farmacología , Administración Oral , Animales , Etanolaminas/administración & dosificación , Corazón/efectos de los fármacos , Corazón/efectos de la radiación , Microsomas/efectos de los fármacos , Microsomas/metabolismo , Microsomas/efectos de la radiación , Microsomas Hepáticos/efectos de los fármacos , Microsomas Hepáticos/metabolismo , Microsomas Hepáticos/efectos de la radiación , Especificidad de Órganos , Protectores contra Radiación/administración & dosificación , Ratas , Ratas Wistar , Ácidos Esteáricos/administración & dosificación , Irradiación Corporal Total , Rayos XRESUMEN
The radioprotective effect of the root extract of Glycyrrhiza glabra L on lipid peroxidation in rat liver microsomes and plasmid pBR322 DNA was investigated. The extract was found to protect microsomal membranes, as evident from reduction in lipid peroxidation, and could also protect plasmid DNA from radiation-induced strand breaks.
Asunto(s)
Daño del ADN/efectos de los fármacos , Glycyrrhiza , Membranas Intracelulares/efectos de los fármacos , Microsomas/efectos de los fármacos , Extractos Vegetales/farmacología , Animales , Daño del ADN/efectos de la radiación , Rayos gamma , Técnicas In Vitro , Membranas Intracelulares/efectos de la radiación , Masculino , Microsomas/efectos de la radiación , Raíces de Plantas/química , Plásmidos/efectos de los fármacos , Plásmidos/efectos de la radiación , Ratas , Ratas WistarRESUMEN
The properties of microsomal membranes in spring wheat leaves (Triticum aestivum L. cv. Ganlong No. 92-005) exposed to (0) control, 8.64 (T1) and 11.2 kJ m(-2) day(-1) (T2) biologically effective UV-B irradiation (UV-B(BE)) were studied under greenhouse conditions. These irradiance levels correspond to a decrease in the stratospheric ozone of approximately 12.5 and 20%, respectively, for a clear solstice day at Lanzhou (36.04 degrees N, 1550 m), China. Compared with controls, the content of malondialdehyde (MDA) increased by 70.8% in T1 and 83.8% in T2 on the 7th day of the radiation, and the IUFA (index of unsaturated fatty acids) decreased, indicating peroxidation of lipid acids. Simultaneously, a drastic decrease of phospholipid content after 21 days and an increase of membrane lipid microviscosity on UV-B irradiation were also found, suggesting a reduction in the fluidity of membrane lipids. Ethylene emission by the microsomal membrane, in the presence of exogenous 1-aminocyclopropane-1-carboxylic acid was higher in the wheat seedlings after 7, 14 and 21 days' irradiation than in the controls. These changes were correlated with a rise in lipoxygenase activity. Membrane-bound enzymes (Ca2+ -ATPase and Mg2+ -ATPase) were promoted by UV radiation in the first 7 days and significantly decreased after 14 and 21 days' treatment in comparison to control. Our results suggest that UV-B radiation may cause changes in structural complexity and function of microsomal membranes in spring wheat leaves.
Asunto(s)
Membranas Intracelulares/efectos de la radiación , Microsomas/efectos de la radiación , Triticum/efectos de la radiación , Rayos Ultravioleta , Relación Dosis-Respuesta en la Radiación , Etilenos/metabolismo , Membranas Intracelulares/metabolismo , Membranas Intracelulares/ultraestructura , Malondialdehído/análisis , Lípidos de la Membrana/metabolismo , Proteínas de la Membrana/efectos de los fármacos , Proteínas de la Membrana/metabolismo , Microsomas/metabolismo , Microsomas/ultraestructura , Fosfolípidos/metabolismo , Reguladores del Crecimiento de las Plantas/metabolismo , Hojas de la Planta/efectos de la radiación , Hojas de la Planta/ultraestructura , Estaciones del Año , Factores de Tiempo , Triticum/fisiologíaRESUMEN
The widely distributed xanthine oxidoreductase (XOR) system has been shown to be modulated upon exposure of animals to ionizing radiation through the conversion of xanthine dehydrogenase (XDH) into xanthine oxidase (XO). In the present work, radiomodification of the XOR system by phenylmethylsulfonyl fluoride (PMSF) and dithiothreitol (DTT) was examined using female Swiss albino mice which were irradiated with gamma rays at a dose rate 0.023 Gy s(-1). PMSF, a serine protease inhibitor, and DTT, the sulfhydryl reagent, were administered intraperitoneally prior to irradiation. The specific activities of XDH and XO as well as the XDH/XO ratio and the total activity (XDH+XO) were determined in the liver of the mice. The inhibition of XO activity, restoration of XDH activity, and increase in the XDH/XO ratio upon administration of PMSF were suggestive of irreversible conversion of XDH into XO mediated through serine proteases. The biochemical events required for the conversion were probably initiated during the early phase of irradiation, as the treatment with PMSF immediately after irradiation did not have a modulatory effect. Interestingly, DTT was not effective in modulating radiation-induced changes in the XOR system or oxidative damage in the liver of mice. The DTT treatment resulted in inhibition of the release of lactate dehydrogenase. However, the protection appears to be unrelated to the formation of TBARS. On the other hand, the presence of PMSF during irradiation inhibited radiation-induced oxidative damage and radiation-induced increases in the specific activity of lactate dehydrogenase. These findings suggest that a major effect of ionizing radiation is irreversible conversion of xanthine to xanthine oxidase.
Asunto(s)
Ditiotreitol/farmacología , Hígado/efectos de la radiación , Fluoruro de Fenilmetilsulfonilo/farmacología , Xantina Deshidrogenasa/efectos de la radiación , Xantina Oxidasa/efectos de la radiación , Animales , Inhibidores Enzimáticos/farmacología , Femenino , Radicales Libres/metabolismo , Glutatión/metabolismo , L-Lactato Deshidrogenasa/metabolismo , Hígado/enzimología , Ratones , Microsomas/efectos de los fármacos , Microsomas/efectos de la radiación , Xantina Deshidrogenasa/efectos de los fármacos , Xantina Deshidrogenasa/metabolismo , Xantina Oxidasa/efectos de los fármacos , Xantina Oxidasa/metabolismoRESUMEN
Induction of heme oxygenase 1 transcription and enzymatic activity is a common response after exposure of cells to various forms of oxidative stress including ultraviolet A radiation (UVA) and hydrogen peroxide. We now show that UVA irradiation or hydrogen peroxide treatment of human skin fibroblasts leads to an immediate release of the heme oxygenase substrate, heme, from microsomal hemeproteins. The release of heme by UVA apparently involves cyclooxygenase activity because it is inhibited by the cyclooxygenase inhibitor indomethacin. We also demonstrate a high degree of correlation between the amount of heme released and the degree of subsequent induction of heme oxygenase 1 transcription following UVA and hydrogen peroxide treatment. We propose that release of heme from microsomal hemeproteins determines the degree of induction of heme oxygenase 1 transcription in human fibroblasts after oxidative stress.
Asunto(s)
Hemo Oxigenasa (Desciclizante)/genética , Hemo/metabolismo , Hemoproteínas/metabolismo , Peróxido de Hidrógeno/farmacología , Microsomas/metabolismo , Prostaglandina-Endoperóxido Sintasas/metabolismo , Transcripción Genética , Rayos Ultravioleta , Hidroxitolueno Butilado/farmacología , Línea Celular , Inhibidores de la Ciclooxigenasa/farmacología , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Fibroblastos/efectos de la radiación , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Regulación Enzimológica de la Expresión Génica/efectos de la radiación , Hemo Oxigenasa (Desciclizante)/biosíntesis , Hemo-Oxigenasa 1 , Humanos , Indometacina/farmacología , Cinética , Proteínas de la Membrana , Microsomas/efectos de los fármacos , Microsomas/efectos de la radiación , Estrés Oxidativo , Piel/efectos de los fármacos , Piel/metabolismo , Piel/efectos de la radiación , Transcripción Genética/efectos de los fármacos , Transcripción Genética/efectos de la radiaciónRESUMEN
Other laboratories have established that oligosaccharyl transferase (OST) from Saccharomyces cerevisiae can be purified as a protein complex containing eight different subunits. To identify the OST subunit that recognizes the peptide sites that can be glycosylated, we developed photoaffinity probes containing a photoreactive benzophenone derivative, p-benzoylphenylalanine (Bpa), as part of an 125I-labeled peptide that could be expected to be glycosylated. We found that Asn-Bpa-Thr peptides served as substrates for OST and that photoactivation of these probes in the presence of microsomes abolished the OST activity. Photoactivation of 125I-labeled Asn-Bpa-Thr in the presence of microsomes resulted in specific covalent labeling of a protein doublet of molecular mass 62 and 64 kDa. By carrying out the photoactivation of the probe using microsomes containing epitope-tagged Ost1p, we demonstrated that the 125I-labeled protein was Ost1p. Radiolabeling of this protein was dependent on irradiation at 350 nm. No labeling was detected using a probe containing Ala instead of Thr as the third amino acid residue. We conclude that Ost1p is the subunit of the OST complex that recognizes the peptide sites in the nascent chains that are destined to be glycosylated.
Asunto(s)
Proteínas Fúngicas/metabolismo , Hexosiltransferasas , Proteínas de la Membrana , Oligopéptidos/metabolismo , Saccharomyces cerevisiae/enzimología , Transferasas/metabolismo , Secuencia de Bases , Secuencia de Consenso , Cartilla de ADN , Proteínas Fúngicas/química , Glicosilación , Microsomas/enzimología , Microsomas/efectos de la radiación , Oligopéptidos/química , Fotoquímica , Especificidad por Sustrato , Transferasas/químicaRESUMEN
The antioxidative effects of grape seed proanthocyanidins (GSPC) were studied in three in-vitro models in which polyunsaturated fatty acids (PUFAs) in aqueous solution and mice liver or brain microsomes were used as oxidative substrates, and UV-C irradiation as the pro-oxidant system. Analysis of UV-C induced lipid peroxidation was carried out by two methods: gas liquid chromatography of residual PUFAs and release of thiobarbituric acid-reactive substances (TBARs) measured by TBA reaction. Results indicate that PUFAs are more radiosensitive when incorporated in single component micelles than in mixed component micelles or microsomes. In every case, PUFA peroxidation was inhibited by low concentrations of GSPC (2 mg/L) while epigallocatecin (EGC) and epigallocatechin gallate (EGCG) monomers, at an equivalent level of epicatechin, exhibited no efficacy in our experimental conditions. This latter effect might be explained by a synergistic action of flavan-3-ol monomers, dimers and oligomers contained in the grape seed extract.
Asunto(s)
Antocianinas/farmacología , Antioxidantes/farmacología , Catequina/análogos & derivados , Ácidos Grasos Insaturados/química , Peroxidación de Lípido/efectos de los fármacos , Rosales/química , Semillas/química , Animales , Encéfalo/efectos de los fármacos , Encéfalo/metabolismo , Encéfalo/efectos de la radiación , Flavonoides/química , Flavonoides/farmacología , Ácido Gálico/química , Ácido Gálico/farmacología , Técnicas In Vitro , Peroxidación de Lípido/efectos de la radiación , Ratones , Microsomas/efectos de los fármacos , Microsomas/metabolismo , Microsomas/efectos de la radiación , Microsomas Hepáticos/efectos de los fármacos , Microsomas Hepáticos/metabolismo , Microsomas Hepáticos/efectos de la radiación , Soluciones , Rayos Ultravioleta , Agua/químicaRESUMEN
The microsomal triglyceride transfer protein (MTP) is a heterodimeric lipid transfer protein that is required for the assembly and secretion of apolipoprotein B (apoB)-containing lipoproteins. A key unresolved question is whether the MTP-mediated step is rate limiting. To address this, a unique experimental strategy was used that allowed the in situ modulation and measurement of MTP triglyceride transfer activity. In order to accomplish this, an irreversible photoaffinity inhibitor, BMS-192951, was designed and synthesized. When incubated with purified MTP and irradiated with UV light at 360 nm, BMS-192951 inhibits triglyceride transfer by covalently binding to the protein. HepG2 cells were treated with either increasing concentrations of BMS-192951 (0-15 microM) with 5 min of ultraviolet irradiation, or 3.0 microM BMS-192951 with various lengths (0-15 min) of ultraviolet irradiation. Microsomal extracts were prepared exhaustively dialyzed to remove unbound inhibitor, and assayed for MTP-mediated triglyceride transfer activity. BMS-192951 was shown to reduce MTP activity in both a dose- and UV exposure time-dependent fashion. Measurement of apoB concentration in the media showed that apoB secretion was reduced in proportion to the in situ inhibition of MTP activity, while no change was observed in apoA-I secretion. Experiments performed in McArdle RH-7777 rat hepatoma cells and primary rat hepatocytes gave nearly identical results; the decrease in apoB secretion was proportional to the decrease in MTP activity. These results indicate that MTP-mediated lipid transfer is limiting in the assembly and secretion of apoB-containing lipoproteins in hepatic cells under the conditions tested.
Asunto(s)
Apolipoproteínas B/biosíntesis , Proteínas Portadoras/metabolismo , Microsomas Hepáticos/metabolismo , Marcadores de Afinidad/farmacología , Animales , Carcinoma Hepatocelular/metabolismo , Bovinos , Células Cultivadas , Cisteína/metabolismo , Humanos , Indoles/química , Indoles/farmacología , Isoindoles , Cinética , Hígado/efectos de los fármacos , Hígado/metabolismo , Hígado/efectos de la radiación , Neoplasias Hepáticas/metabolismo , Metionina/metabolismo , Microsomas/efectos de los fármacos , Microsomas/metabolismo , Microsomas/efectos de la radiación , Microsomas Hepáticos/efectos de los fármacos , Microsomas Hepáticos/efectos de la radiación , Piperidinas/química , Piperidinas/farmacología , Ratas , Radioisótopos de Azufre , Triglicéridos/metabolismo , Células Tumorales Cultivadas , Rayos UltravioletaRESUMEN
Folding of polypeptides emerging from the protein translocase in the membrane of mammalian microsomes was analyzed after synthesis of corresponding precursor proteins in a mammalian translation system. Firefly luciferase was used as a model protein; the corresponding hybrid precursor contained the preprolactin signal peptide. The rates and efficiencies of folding of luciferase in microsomes were compared with those of folding of luciferase in the cytosol. Furthermore, folding of luciferase in microsomes was compared with that in proteoliposomes, i.e. in the absence of luminal molecular chaperones and folding catalysts. Folding in microsomes was less efficient compared with folding in the cytosol. Folding in the absence of luminal proteins was more efficient compared with folding in their presence and identical to folding in the cytosol. Thus, firefly luciferase emerging from translocase can efficiently fold to its native conformation without chaperoning by any luminal proteins. There may be molecular chaperones present in the microsomal membrane that can efficiently substitute for the cytosolic chaperone machinery comprising Hsp40, Hsp60, and Hsp70 with respect to folding of firefly luciferase.
Asunto(s)
Escarabajos/enzimología , Luciferasas/metabolismo , Microsomas/metabolismo , Chaperonas Moleculares , Adenosina Trifosfato/análogos & derivados , Adenosina Trifosfato/farmacología , Secuencia de Aminoácidos , Animales , Azidas/farmacología , Transporte Biológico , Catálisis , Ciclosporina/farmacología , Perros , Microsomas/efectos de los fármacos , Microsomas/efectos de la radiación , Datos de Secuencia Molecular , Pliegue de Proteína , Proteolípidos/metabolismo , Tacrolimus/farmacología , Rayos UltravioletaRESUMEN
There is increasing evidence that oxygen free radicals (OFR) are involved in cerebral ischaemia-reperfusion injury, possibly via a modulation of Na+,K(+)-ATPase activity, one of the major membrane pumps responsible for ionic homeostasis. We measured OFR-mediated modulation of this enzymatic activity and examined the roles of lipid and/or protein alterations. Using mouse brain microsomes exposed to UV-C irradiation, our results show a good correlation between activity inhibition and lipoperoxidation estimated by PUFA loss as well as malondialdehyde production. The protective effect of thiourea (OH scavenger) and the lack of effect noted with DTT (thiol protector) suggest that the functionality of the Na+,K(+)-ATPase is altered by perturbation of membrane integrity rather than by a structural alteration of the protein itself.
Asunto(s)
Adenosina Trifosfato/farmacología , Encéfalo/enzimología , Microsomas/enzimología , Especies Reactivas de Oxígeno/metabolismo , ATPasa Intercambiadora de Sodio-Potasio/metabolismo , Animales , Encéfalo/efectos de los fármacos , Encéfalo/efectos de la radiación , Encéfalo/ultraestructura , Ditiotreitol/farmacología , Depuradores de Radicales Libres/farmacología , Radicales Libres , Peroxidación de Lípido/efectos de los fármacos , Masculino , Ratones , Microsomas/efectos de los fármacos , Microsomas/efectos de la radiación , Proteínas del Tejido Nervioso/metabolismo , Fármacos Neuroprotectores/farmacología , Oxidación-Reducción , Tiourea/farmacología , Rayos UltravioletaRESUMEN
The effects of 50 Hz, 1.2 mT magnetic fields (MFs) were tested on hepatic monooxygenase enzymes of basal and beta-naphthoflavone-phenobarbital-preinduced rats and mice. An inductive effect on cytochrome P-450 level and on some enzymatic cytochrome P-450-dependent activities was observed in basal mice after MF exposure. Enzymatic activities in preinduced mice and rats were reduced by MFs, the degree of reduction depending on the enzyme. A specific inhibitory effect was determined in some of the assayed activities and in the relative peculiar P-450 isoforms detected by Western blot analysis.
Asunto(s)
Campos Electromagnéticos , Hígado/enzimología , Magnetismo , Oxidorreductasas/efectos de la radiación , 7-Alcoxicumarina O-Dealquilasa/efectos de los fármacos , 7-Alcoxicumarina O-Dealquilasa/efectos de la radiación , Anilina Hidroxilasa/efectos de los fármacos , Anilina Hidroxilasa/efectos de la radiación , Animales , Benzoflavonas/farmacología , Western Blotting , Citocromo P-450 CYP2B1 , Citocromo P-450 CYP2E1 , Sistema Enzimático del Citocromo P-450/efectos de los fármacos , Sistema Enzimático del Citocromo P-450/efectos de la radiación , Isoenzimas/efectos de los fármacos , Isoenzimas/efectos de la radiación , Hígado/efectos de los fármacos , Hígado/efectos de la radiación , Ratones , Microsomas/efectos de los fármacos , Microsomas/enzimología , Microsomas/efectos de la radiación , Oxidorreductasas/efectos de los fármacos , Oxidorreductasas N-Desmetilantes/efectos de los fármacos , Oxidorreductasas N-Desmetilantes/efectos de la radiación , Oxidorreductasas O-Demetilantes/efectos de los fármacos , Oxidorreductasas O-Demetilantes/efectos de la radiación , Fenobarbital/farmacología , Ratas , Ratas Wistar , beta-naftoflavonaRESUMEN
In recent years green tea has been shown to afford protection against chemical- and photo-carcinogenesis in several animal tumor bioassay systems. It has been suggested that the wide range of anticarcinogenic properties of green tea may be due to the antioxidant effect of epicatechins present therein. In this study, we assessed whether these epicatechin derivatives (ECDs)--namely (-)-epigallocatechin (EGC), (-)-epicatechin gallate (ECG), (-)-epigallocatechin-3-gallate (EGCG) and (-)-epicatechin (EC) inhibit spontaneous and photo-enhanced lipid peroxidation (LPO) in mouse epidermal microsomes. Our data indicate that significant inhibition (significance levels for P, < 0.05 to < 0.0001) was evident by EGCG, EGC and ECG in Fe3+/ADP supported LPO. Interestingly each of these epicatechin derivatives was also effective in inhibiting photo-enhanced LPO generated by incubating epidermal microsomes in the presence of silicon phthalocyanine and 650 nm irradiation. However, at equimolar basis, EGCG, which is also the major constituent in GTP, showed maximum inhibitory effects compared to other ECDs. Taken together, our results provide the direct evidence for the antioxidant property of ECDs, and suggest that such an effect may contribute towards anticarcinogenic (specifically anti-skin tumor) promoting effects of green tea.
Asunto(s)
Catequina/análogos & derivados , Catequina/farmacología , Epidermis/metabolismo , Peroxidación de Lípido/efectos de los fármacos , Microsomas/metabolismo , Silanos , Piel/metabolismo , Té , Animales , Epidermis/efectos de los fármacos , Epidermis/efectos de la radiación , Indoles/farmacología , Luz , Peroxidación de Lípido/efectos de la radiación , Ratones , Ratones Endogámicos , Microsomas/efectos de los fármacos , Microsomas/efectos de la radiación , Compuestos de Organosilicio/farmacología , Fármacos Fotosensibilizantes/farmacología , Piel/efectos de los fármacos , Piel/efectos de la radiación , Relación Estructura-ActividadRESUMEN
For evaluation of the possibility of the appearance of autoimmune thyroiditis in children and juveniles lived in the areas of Kaluga Province [correction of region] suffered from the Chernobyl accident the content of antibodies to human thyroid microsomal antigen was investigated. Percentage of positive sera varied from 4.8% to 1.2% during 6 years. There is significant difference in the frequency of the antibody appearance between persons suffered from radioactive iodine and unsuffered ones. Correlation between levels of antimicrosomal antibodies and radiation dose obtained from incorporated radioactive iodine was not estimated.
Asunto(s)
Accidentes de Trabajo , Contaminación Radiactiva del Aire/efectos adversos , Autoanticuerpos/sangre , Autoantígenos/inmunología , Exposición a Riesgos Ambientales/efectos adversos , Microsomas/inmunología , Reactores Nucleares , Centrales Eléctricas , Glándula Tiroides/inmunología , Adolescente , Autoanticuerpos/efectos de la radiación , Autoantígenos/efectos de la radiación , Niño , Relación Dosis-Respuesta en la Radiación , Humanos , Microsomas/efectos de la radiación , Federación de Rusia , Glándula Tiroides/efectos de la radiación , Tiroiditis Autoinmune/etiología , Tiroiditis Autoinmune/inmunología , UcraniaRESUMEN
A brief literature review shows that ionizing radiation in biological membranes and in pure lipid membranes causes malondialdehyde formation, indicating lipid peroxidation processes. With respect to membrane fluidization by ionizing radiation, in pure lipid membranes rigidization effects are always reported, whereas contradictory results exist for biological membranes. Starting from the assumption that membrane proteins at least partly compensate for radiation effects leading to a rigidization of membrane lipid regions, pig liver microsomes, as a representative protein-rich intracellular membrane system, were irradiated with X-rays or UV-C with doses up to 120 Gy at a dose rate of 0.67 Gy min-1 and up to 0.73 J cm-2 at an exposure rate of 16.2 mJ cm-2 min-1, respectively. For both irradiation types a weak but significant positive correlation between malondialdehyde formation and membrane fluidity is revealed throughout the applied dose ranges. We conclude that the membraneous protein lipid interface increases its fluidity under radiation conditions. Also, thymocyte ghosts showed an increased fluidity after X-ray irradiation. Fluidity measurements were performed by the pyrene excimer method.
Asunto(s)
Fluidez de la Membrana/efectos de la radiación , Microsomas/efectos de la radiación , Linfocitos T/efectos de la radiación , Animales , Relación Dosis-Respuesta en la Radiación , Peroxidación de Lípido , Ratones , Porcinos , Rayos Ultravioleta , Rayos XRESUMEN
The principal aim of the study was to investigate the effect of low-dose gamma-irradiation on lipid peroxidation (LPO) in murine bone marrow. To this end, the degree of LPO in suspensions of microsomes of murine bone marrow cells (BMC) was determined in terms of malondialdehyde (MDA) formation after whole-body or in vitro exposure to various doses of gamma-radiation. These effects were compared to some extent with similar effects in liver and spleen preparations. As to the effect of gamma-irradiation on LPO in BMC, the response depends on the dose level and on whether whole-body or in vitro exposures are involved. Whole-body irradiation did not result in an increase in LPO in BMC microsomes, even at such high doses as 15 Gy, although hepatic microsomes showed a marked increase. In contrast, in vitro irradiation of BMC microsomes with 0.1, 10 and 50 Gy brought about an increase in LPO. This increase was already significant (P < 0.05) at 0.1 Gy following a post-irradiation incubation and substantial at 50 Gy, even without subsequent incubation. The results show that low doses of gamma-irradiation are able to induce an elevation of LPO in murine BMC microsomes, but only after in vitro irradiation. In the case of whole-body irradiation cellular radical scavengers and other metabolic reactions may prevent a measurable increase in LPO. This is partly illustrated by the case of vitamin-E deficiency, where a substantial increase in LPO in BMC microsomes is observed even without gamma-irradiation in comparison with euvitaminotic mice because normally occurring radicals are not scavenged sufficiently.
Asunto(s)
Médula Ósea/efectos de la radiación , Peroxidación de Lípido/efectos de la radiación , Microsomas/efectos de la radiación , Animales , Médula Ósea/metabolismo , Femenino , Radicales Libres , Rayos gamma , Ratones , Microsomas/metabolismo , Irradiación Corporal TotalRESUMEN
To determine the size of the functional catalytic unit of prostaglandin endoperoxide (prostaglandin H) synthase, radiation inactivation experiments were performed. Both microsomes from ovine seminal vesicles and purified enzyme were irradiated with 10 MeV electrons. The enzymic activities of prostaglandin H synthase, cyclooxygenase and peroxidase, showed mono-exponential inactivation curves dependent on radiation dose, indicating molecular masses of approximately 72 kDa. The enzyme in microsomes, in its native environment, as well as in its purified state after solubilisation with nonionic detergent showed identical molecular masses. The results clearly demonstrate that the monomer of the enzyme with an apparent molecular mass of 72 kDa (SDS/PAGE) is the functional unit for catalysis of both activities. Hence the two active sites of cyclooxygenase and peroxidase reside on the same polypeptide chain.