RESUMEN
BACKGROUND: Inflammatory response triggered by high mobility group box-1 (HMGB1) protein and oxidative stress play critical roles in the intestinal injury after severe burn. Sodium butyrate, a histone deacetylase inhibitor, has potential anti-inflammatory properties, inhibiting the expression of inflammatory mediators such as HMGB1 in diverse diseases. This study was designed to investigate the effects of sodium butyrate on severe burn plus delayed resuscitation-induced intestine injury, intestinal expressions of HMGB1 and intracellular adhesion molecule-1 (ICAM-1), oxidative stress, and signal transduction pathway changes in rats. MATERIALS AND METHODS: Fifty-six Sprague-Dawley rats were divided into 3 groups randomly: (1) sham group, animals underwent sham burn; (2) burn group, rats subjected to full-thickness burns of 30% total body surface area (TBSA) and received 2ml/kg/TBSA lactated Ringer solution for resuscitation at 6, 12, and 36h after burn injury; (3) burn plus sodium butyrate (burn+SB) group, animals received burn injury and lactated Ringer solution with sodium butyrate inside for resuscitation in the same manner. Diamine oxidase (DAO) concentration in plasma was measured by enzyme-linked immunosorbent assay. Intestinal fatty acid binding protein (I-FABP) and ICAM-1 expressions in the intestine were analyzed by immunohistochemical method. HMGB1 and p38 mitogen-activated protein kinase (MAPK) expressions in the intestine tissues were examined by Western blot. The intestinal concentration of malondialdehyde (MDA) was also determined. RESULTS: Intestinal HMGB1 expression was significantly increased in burn group compared with sham group. Sodium butyrate administration significantly inhibited the HMGB1 expression in the intestine, decreased the DAO concentration in plasma, reduced the intestinal I-FABP expression, and improved the intestinal histologic changes induced by burn injury plus delayed resuscitation. Sodium butyrate treatment also markedly reduced the increase of intestinal ICAM-1 expression and MDA content, and inhibited p38 MAPK activity in the intestine of severely burned rats with delayed resuscitation. CONCLUSIONS: Sodium butyrate inhibits HMGB1 expression which could be attributed to p38 MAPK signal transduction pathway and decreases intestinal inflammatory responses and oxidative stress, thus attenuates burn plus delayed resuscitation-induced intestine injury.
Asunto(s)
Quemaduras/metabolismo , Ácido Butírico/farmacología , Proteína HMGB1/efectos de los fármacos , Antagonistas de los Receptores Histamínicos/farmacología , Íleon/efectos de los fármacos , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Amina Oxidasa (conteniendo Cobre)/efectos de los fármacos , Amina Oxidasa (conteniendo Cobre)/metabolismo , Animales , Superficie Corporal , Quemaduras/patología , Proteínas de Unión a Ácidos Grasos , Fluidoterapia , Proteína HMGB1/metabolismo , Íleon/metabolismo , Íleon/patología , Molécula 1 de Adhesión Intercelular/efectos de los fármacos , Molécula 1 de Adhesión Intercelular/metabolismo , Malondialdehído/metabolismo , Estrés Oxidativo/efectos de los fármacos , Ratas , Resucitación , Lactato de RingerRESUMEN
BACKGROUND: Semicarbazide-sensitive amine oxidase (SSAO; EC; 1.4.3.6.) has widespread tissue distribution, and the physiological role of SSAO is quite well known through its involvement in several pathological states. AIMS: The present study examined modulators of SSAO which might be present in the rat heart cytosol and looked for changes in SSAO modulatory activity. METHODS: An endogenous inhibitor of SSAO was separated by gel filtration from 105,000g supernate of T4-treated rat heart cytosol. SSAO inhibition fraction was referred to as "endogenous SSAO inhibitor". RESULTS: The inhibition by this inhibitor was concentration-dependent. Inhibition of SSAO was not enhanced by varying the time of preincubation of the enzyme, indicating reversible inhibition of SSAO. The molecular weight of this inhibitor was estimated to be 1000-1100 by gel filtration. The isoelectric point (pI) value was determined to be 4.8 isoelectric focusing. This inhibitor was found to be heat-stable and resistant to protease treatment. SSAO inhibition activity was much lower in the cytosol of thyroidectomized, non-T4-treated rats than T4-treated rats, suggesting that this inhibitor was induced by thyroid hormone T4. SSAO activity in rat heart might be regulated by the level of this inhibitor. CONCLUSION: These results suggest the presence of SSAO inhibitor in T4-treated rat cytosol and that the level of this inhibitor is regulated by thyroid hormone.
Asunto(s)
Amina Oxidasa (conteniendo Cobre)/metabolismo , Citosol/metabolismo , Miocardio/metabolismo , Miocitos Cardíacos/metabolismo , Tiroxina/farmacología , Amina Oxidasa (conteniendo Cobre)/efectos de los fármacos , Amina Oxidasa (conteniendo Cobre)/economía , Animales , Modelos Animales de Enfermedad , Masculino , Miocitos Cardíacos/patología , Ratas , Ratas WistarRESUMEN
With better understanding of the molecular mechanisms underpinning chronic kidney disease, the roles of inflammation and fibrosis are becoming increasingly inseparable. The progression of renal disease is characterized by pathomorphological changes that consist of early inflammatory responses followed by tubulointerstitial fibrosis, tubular atrophy, and glomerular and vascular sclerosis. Currently available therapies that reduce hypertension, proteinuria, hyperglycemia, and interruption of the renin-angiotensin-aldosterone system are at best only partially effective. Hence, there remains a need to explore agents targeting nonrenin-angiotensin-aldosterone system pathways. In this review, we discuss mechanistic aspects in the physiological and pathological role of semicarbazide-sensitive amine oxidase, a protein enzyme involved in cellular trafficking and inflammation, with respect to the kidney. We explore the evidence for the use of semicarbazide-sensitive amine oxidase inhibitors as potential agents in renal fibrosis to delay the onset and progression of chronic kidney disease.
Asunto(s)
Amina Oxidasa (conteniendo Cobre)/fisiología , Insuficiencia Renal Crónica/fisiopatología , Amina Oxidasa (conteniendo Cobre)/antagonistas & inhibidores , Amina Oxidasa (conteniendo Cobre)/efectos de los fármacos , Inhibidores Enzimáticos/farmacología , Inhibidores Enzimáticos/uso terapéutico , Humanos , Riñón/patología , Riñón/fisiopatología , Insuficiencia Renal Crónica/prevención & control , Sistema Renina-Angiotensina/fisiologíaRESUMEN
BACKGROUND: The specific purpose of this study was to investigate the effects of medium-chain triglycerides (MCTs) on intestinal cell proliferation and mucous secretion of the small intestine in the rat. METHODS: Rats were fed chow diet and given MCTs or the same weight of corn oil (5 g/kg per day) by gavage daily for 2 weeks, and then tissue samples of the small intestines were harvested. Leptin concentration in the small intestine was measured. Cell proliferation and apoptosis in the small intestine was determined by immunohistochemistry. Diamine oxidase (DAO) activity was measured by colorimetric assay. RESULTS: In rats fed only chow diet (normal rats), the number of goblet cells per villi was 14.2 +/- 0.75 in the jejunum and 15.2 +/- 1.12 in the ileum. The number of goblet cells increased significantly in rats given MCTs compared with rats given corn oil or normal rats. Ki-67-positive cells were detected on the entire villi and the crypts in the small intestine. Furthermore, the proliferative index and the apoptotic index were also significantly greater in rats given MCTs than rats given corn oil or normal rats. Moreover, DAO activity and leptin concentration in the small intestine were significantly greater in rats given MCTs than rats given corn oil or normal rats. CONCLUSIONS: MCTs enhance cell proliferation of the intestinal epithelium and mucous secretion from goblet cells in the small intestine. These effects may protect the gut in patients suffering from inflammatory bowel disease or enterogenous infection.
Asunto(s)
Proliferación Celular/efectos de los fármacos , Mucosa Intestinal/efectos de los fármacos , Moco/efectos de los fármacos , Triglicéridos/farmacología , Amina Oxidasa (conteniendo Cobre)/efectos de los fármacos , Amina Oxidasa (conteniendo Cobre)/metabolismo , Animales , Apoptosis/efectos de los fármacos , Células Caliciformes/efectos de los fármacos , Células Caliciformes/metabolismo , Íleon/efectos de los fármacos , Íleon/metabolismo , Inmunohistoquímica , Mucosa Intestinal/metabolismo , Yeyuno/efectos de los fármacos , Yeyuno/metabolismo , Antígeno Ki-67/metabolismo , Leptina/metabolismo , Masculino , Moco/metabolismo , Ratas , Ratas Sprague-DawleyRESUMEN
The human mast cell line (HMC-1(560, 816)) was used to study the effect of the tyrosine kinase inhibitor STI571 (Glivec) on exocytosis, intracellular Ca(2+) and pH changes, because STI571 inhibits the proliferation of HMC-1(560) and induces its apoptosis. This drug does not have these effects on HMC-1(560, 816). Exocytosis in HMC-1(560, 816) cells can be stimulated by alkalinisation with NH(4)Cl as well as with ionomycin. Surprisingly 24-h pre-incubation with STI571 decreases spontaneous histamine release of HMC-1(560, 816) cells, but increases the histamine response after alkalinisation and not after ionomycin-stimulation. After addition of NH(4)Cl, pH(i) has a higher increase in STI571 pre-incubated cells, without changing intracellular Ca(2+) concentration. Activation of PKC in combination with tyrosine kinase inhibition increases also histamine release in HMC-1(560, 816) cells. Strangely, STI571 pre-incubated cells with PKC inhibited by rottlerin show the same effects. In these cells, cytosolic pH increases more than in control cells. This is the first report of STI571 effect in HMC-1(560, 816) cells. It seems that different pathways modulate signals for proliferation and exocytosis. STI571 does not only inhibit KIT TyrK, but may also influence cytosolic pH after alkalinisation in both cell lines, HMC-1(560) and HMC-1(560, 816), and this ends in induced histamine release. This work is important since HMC-1(560, 816) cells are reported in 80% of aggressive systemic mastocytosis cases and the understanding of some signalling pathways involved in mast cell response could facilitate drug targeting.
Asunto(s)
Amina Oxidasa (conteniendo Cobre)/efectos de los fármacos , Apoptosis/efectos de los fármacos , Diferenciación Celular/fisiología , Liberación de Histamina/fisiología , Mastocitos/metabolismo , Piperazinas/farmacología , Pirimidinas/farmacología , Acetofenonas/farmacología , Álcalis/farmacología , Benzamidas , Benzopiranos/farmacología , Calcio/química , Calcio/metabolismo , Línea Celular , Proliferación Celular , Citoplasma/química , Sistemas de Liberación de Medicamentos , Inhibidores Enzimáticos/farmacología , Exocitosis/efectos de los fármacos , Humanos , Concentración de Iones de Hidrógeno , Mesilato de Imatinib , Mastocitosis Sistémica , Proteína Quinasa C/metabolismo , Inhibidores de Proteínas Quinasas/farmacología , Proteínas Tirosina Quinasas/farmacología , Transducción de SeñalRESUMEN
Semicarbazide-sensitive amine oxidase (SSAO) is a multifunctional enzyme with different biological roles that depend on the tissue where it is expressed. Because SSAO activity is altered in several pathological conditions, we were interested in studying the possible regulation of the human enzyme activity. It has been previously reported that SSAO activity is increased in the presence of Dulbecco's modified Eagle medium (DMEM) in vitro. The aim of the present work was to investigate the effects of the different constituents of DMEM on human SSAO activity. We found that sodium bicarbonate was the only component able to mimic the enhancement of both human aorta and plasma SSAO activity in vitro, suggesting a possible physiological role of bicarbonate as an intrinsic modulator of the human enzyme. Failure to take this activating effect into account could also result in inaccuracies in the reported tissue activities of this enzyme.
Asunto(s)
Amina Oxidasa (conteniendo Cobre)/efectos de los fármacos , Moléculas de Adhesión Celular/metabolismo , Membrana Celular/metabolismo , Medios de Cultivo , Bicarbonato de Sodio/farmacología , Amina Oxidasa (conteniendo Cobre)/metabolismo , Northern Blotting , Activación Enzimática , Humanos , SolubilidadRESUMEN
Tranylcypromine (TCP), an amphetamine, is a reversible inhibitor of copper-containing amine oxidases. We have solved the structure of the complex of TCP with the amine oxidase from E. coli (ECAO) and shown that only the (+)-enantiomer of TCP binds. Kinetic studies on 2-phenylethylamine and TCP binding to wild-type ECAO and mutational variants fully support the model in which binding of the protonated amine is the first step in the catalytic cycle. Hydrazines are irreversible inhibitors of copper-containing amine oxidases. Binding of hydrazines leads to an adduct ("Adduct 1") with a chromophore at 430 nm which converts at higher pH to another adduct ("Adduct 2") with a chromophore at 520 nm. We have determined the structures of Adduct 1 and 2 for 2-hydrazinopyridine reacted with ECAO. It has been found that Adduct 1 corresponds to the hydrazone and azo tautomers whilst Adduct 2 corresponds to the azo tautomer coordinated to the active site copper. The implications of these results in developing more specific drugs are discussed.
Asunto(s)
Amina Oxidasa (conteniendo Cobre)/química , Anfetaminas/química , Dominio Catalítico/efectos de los fármacos , Hidrazinas/química , Tranilcipromina/química , Amina Oxidasa (conteniendo Cobre)/efectos de los fármacos , Amina Oxidasa (conteniendo Cobre)/metabolismo , Anfetaminas/metabolismo , Anfetaminas/farmacología , Sitios de Unión/efectos de los fármacos , Sitios de Unión/fisiología , Dominio Catalítico/fisiología , Cobre/química , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/metabolismo , Inhibidores Enzimáticos/farmacología , Hidrazinas/metabolismo , Hidrazinas/farmacología , Isomerismo , Conformación Molecular , Estructura Molecular , Inhibidores de la Monoaminooxidasa/química , Inhibidores de la Monoaminooxidasa/metabolismo , Inhibidores de la Monoaminooxidasa/farmacología , Unión Proteica/efectos de los fármacos , Unión Proteica/fisiología , Piridonas/química , Piridonas/metabolismo , Piridonas/farmacología , Tranilcipromina/metabolismo , Tranilcipromina/farmacologíaRESUMEN
Semicarbazide-sensitive amine oxidase/vascular adhesion protein-1 (SSAO) is believed to be a bifunctional membrane protein. It is localized extracellularly and preferentially oxidizes short chain primary amines to aldehydes, hydrogen peroxide and ammonia, but also functions as an adhesion molecule, which is involved in leukocyte migration. Serum SSAO activity is increased in diabetic patients and animals and the aldehydes formed in the enzyme reaction may contribute to vascular damage. However, administration of exogenous substrates has been shown to improve glucose tolerance and reduce hyperglycaemia in diabetic animals. Hydrogen peroxide and/or its vanadate complexes have been suggested responsible for these effects. Streptozotocin induced diabetic rats were treated with benzylamine (BZA) +/- vanadate (V) or insulin. In contrast to insulin, BZA + V treatment did not reduce HbA(1C) levels. However, it reduced the elevated serum SSAO activity, decreased the accumulation of advanced-glycation end products and increased the bioavailability of nitric oxide in diabetic animals, similarly to insulin. BZA alone did not affect any of these parameters.
Asunto(s)
Amina Oxidasa (conteniendo Cobre)/sangre , Bencilaminas/farmacología , Diabetes Mellitus Experimental/tratamiento farmacológico , Diabetes Mellitus Experimental/enzimología , Hipoglucemiantes/farmacología , Insulina/agonistas , Amina Oxidasa (conteniendo Cobre)/efectos de los fármacos , Animales , Bencilaminas/uso terapéutico , Glucemia/efectos de los fármacos , Glucemia/fisiología , Diabetes Mellitus Experimental/fisiopatología , Regulación hacia Abajo/efectos de los fármacos , Regulación hacia Abajo/fisiología , Sinergismo Farmacológico , Quimioterapia Combinada , Hemoglobina Glucada/efectos de los fármacos , Hemoglobina Glucada/metabolismo , Productos Finales de Glicación Avanzada/sangre , Hipoglucemiantes/uso terapéutico , Insulina/metabolismo , Masculino , Óxido Nítrico/metabolismo , Ratas , Ratas Wistar , Vanadatos/farmacología , Vanadatos/uso terapéuticoRESUMEN
Semicarbazide-sensitive amine oxidase (SSAO) also functions as a vascular-adhesion protein (VAP-1). The nature of the target site on lymphocytes to which endothelial-cell SSAO/VAP-1 binds is unknown. We have shown that amino sugars (galactosamine, glucosamine and mannosamine), which are not SSAO substrates, can bind to the enzyme as reversible inhibitors. Thus, they serve as a model system in which to study the interaction process. Binding occurred during substrate (benzylamine) oxidation but not when the amino sugar was incubated, for extended periods, with SSAO alone. These results suggest that one, or more of the products of the SSAO-catalysed amine oxidation might be necessary for the inhibitory process to occur. Two of the reaction products of benzylamine oxidation, benzaldehyde and ammonia were found to have no effect on the inhibition of SSAO by galactosamine. Preincubation of the enzyme with galactosamine plus H(2)O(2) was, however, found to result in time-dependent inhibition. This is not a result of the non-enzymic reaction between H(2)O(2) and the amino sugar, since preincubation of galactosamine with H(2)O(2) alone, for extended periods, did not give rise to an inhibitory species. The amount of exogenously added H(2)O(2) necessary for inhibition was very much greater than that formed during substrate oxidation. These results suggest that the H(2)O(2) formed as a product of the SSAO-catalysed oxidation reaction is more efective in promoting the binding of amino sugars.
Asunto(s)
Amina Oxidasa (conteniendo Cobre)/metabolismo , Aminas/metabolismo , Amino Azúcares/metabolismo , Moléculas de Adhesión Celular/metabolismo , Peróxido de Hidrógeno/metabolismo , Amina Oxidasa (conteniendo Cobre)/química , Amina Oxidasa (conteniendo Cobre)/efectos de los fármacos , Aminas/química , Amino Azúcares/química , Animales , Bencilaminas/química , Bencilaminas/metabolismo , Dominio Catalítico/efectos de los fármacos , Dominio Catalítico/fisiología , Bovinos , Moléculas de Adhesión Celular/química , Moléculas de Adhesión Celular/efectos de los fármacos , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/metabolismo , Inhibidores Enzimáticos/farmacología , Galactosamina/química , Galactosamina/metabolismo , Peróxido de Hidrógeno/química , Peróxido de Hidrógeno/farmacología , Oxidación-Reducción , Unión Proteica/efectos de los fármacos , Unión Proteica/fisiologíaRESUMEN
We and others have observed that substrates for copper-containing amine oxidases cause substrate inhibition at high concentrations. Through use of a novel "pseudoquantitative" rapid equilibrium approach, kinetic analyses with human and bovine enzymes indicate that these effects are consistent with substrates binding to oxidised and reduced enzyme forms. Small cations compete with binding of substrates to oxidised and reduced enzyme, influencing both substrate turnover and substrate inhibition patterns. Cations reduce affinity of the resting bovine enzyme for spermidine, but not benzylamine, indicating that the predominant effect of cations on substrate oxidation results from binding to an anionic site outside the active site. However, binding of cations to the active site of the reduced form of both enzymes attenuates substrate inhibition with both spermidine and benzylamine. Our observations have significant practical implications for researchers assaying kinetic behaviour of these enzymes, and particularly those developing novel inhibitors of human copper-containing amine oxidases.
Asunto(s)
Amina Oxidasa (conteniendo Cobre)/química , Amina Oxidasa (conteniendo Cobre)/metabolismo , Dominio Catalítico/fisiología , Cationes/química , Cobre/química , Amina Oxidasa (conteniendo Cobre)/efectos de los fármacos , Animales , Bencilaminas/química , Bencilaminas/metabolismo , Tampones (Química) , Cationes/farmacología , Bovinos , Moléculas de Adhesión Celular/química , Moléculas de Adhesión Celular/efectos de los fármacos , Moléculas de Adhesión Celular/metabolismo , Humanos , Cinética , Oxidación-Reducción/efectos de los fármacos , Unión Proteica/efectos de los fármacos , Unión Proteica/fisiología , Espermidina/química , Espermidina/metabolismoRESUMEN
Semicarbazide-sensitive amine oxidase (EC 1.4.3.6) acts as a vascular-adhesion protein (VAP-1), mediating the adhesion of lymphocytes to vascular endothelial cells under inflammatory conditions. The relationship between the adhesive and the enzymatic functions of SSAO have not yet been fully defined. Previous studies from this laboratory showed aminohexoses, which were neither substrates nor direct inhibitors of SSAO, bound to the enzyme as reversible inhibitors in the presence of H(2)O(2) generated during substrate oxidation. The possibility that surface L-lysine could act similarly has been investigated in the present study. The presence of L-lysine during the oxidation of benzylamine resulted in time- and dose-dependent inhibition of SSAO activity, in a process that was dependent on the H(2)O(2) formed during benzylamine oxidation. The possible implications of this in terms of the therapeutic uses of lysine are discussed.
Asunto(s)
Amina Oxidasa (conteniendo Cobre)/química , Dominio Catalítico/fisiología , Moléculas de Adhesión Celular/química , Lisina/química , Amina Oxidasa (conteniendo Cobre)/efectos de los fármacos , Amina Oxidasa (conteniendo Cobre)/metabolismo , Animales , Bencilaminas/química , Bencilaminas/metabolismo , Dominio Catalítico/efectos de los fármacos , Adhesión Celular/efectos de los fármacos , Adhesión Celular/fisiología , Moléculas de Adhesión Celular/efectos de los fármacos , Moléculas de Adhesión Celular/metabolismo , Quimiotaxis de Leucocito/fisiología , Relación Dosis-Respuesta a Droga , Células Endoteliales/enzimología , Humanos , Peróxido de Hidrógeno/metabolismo , Cinética , Lisina/metabolismo , Lisina/farmacología , Oxidación-Reducción , Unión Proteica/fisiología , Factores de TiempoRESUMEN
OBJECTIVE: Although the treatment of schizophrenia with many second generation antipsychotics is known to be associated with metabolic changes, such as hyperglycemia or hypercholesterolemia, the underlying mechanisms of these adverse reactions remain unclear. In light of the recent focus on the involvement of semicarbazide-sensitive amine oxidase (SSAO) in glucose metabolism, we investigated SSAO serum activity in schizophrenic patients treated with antipsychotics with the objective of determining a possible link between SSAO and impaired glucose metabolism. METHODS: Blood samples were drawn from 44 schizophrenic patients (24 receiving second generation antipsychotics known to disturb glucose metabolism) on day 1 and day 5 of inpatient treatment. Forty-one healthy adults with no medical condition known to influence SSAO served as controls. RESULTS: Of the 44 schizophrenic patients, the 24 treated with second generation antipsychotics known to disturb glucose metabolism showed significantly lower SSAO serum activity [day 1 (mean +/- SD): 477 +/- 105 mU/L; day 5: 438 +/- 86 mU/L] than the 20 patients treated with other antipsychotics not known to influence glucose metabolism (day 1: 513 +/- 124 mU/L, p = 0.359; day 5: 542 +/- 204 mU/L, p = 0.021) only after 5 days of treatment and compared to healthy controls (526 +/- 142 mU/L, p = 0.021). No differences were observed between schizophrenic patients treated with first generation antipsychotics and the controls. CONCLUSION: We found decreased SSAO serum activity exclusively in schizophrenic patients treated with second generation antipsychotics known to disturb glucose metabolism. In terms of the role of SSAO in glucose metabolism, the observed decrease in SSAO serum activity may be linked to metabolic changes that are known to occur in schizophrenic patients being treated with many second generation antipsychotics.
Asunto(s)
Amina Oxidasa (conteniendo Cobre)/efectos de los fármacos , Antipsicóticos/efectos adversos , Glucosa/metabolismo , Esquizofrenia/tratamiento farmacológico , Adulto , Amina Oxidasa (conteniendo Cobre)/sangre , Amina Oxidasa (conteniendo Cobre)/metabolismo , Antipsicóticos/farmacología , Antipsicóticos/uso terapéutico , Femenino , Humanos , Masculino , Persona de Mediana Edad , Análisis Multivariante , Proyectos PilotoRESUMEN
To clarify whether transition metals are involved in carbonyl stress in diabetic tissues, we observed the effects of a metal chelating agent, trientine (TE) hydrochloride on the levels of methylglyoxal (MG), 3-deoxyglucosone (3-DG), advanced glycation end products, 8-hydroxy-2'-deoxyguanosine (8-OHdG), and polyol pathway metabolites along with semicarbazide-sensitive amine oxidase (SSAO) enzyme activity in lenses from streptozotocin-induced diabetic rats. Lens MG and 3-DG levels were significantly higher in diabetic rats than nondiabetic controls, and TE significantly restored the increase of these compounds. Lens argpyrimidine was also increased in diabetic rats as compared with controls and was significantly reduced by TE. Lens SSAO activity and 8-OHdG were also significantly elevated in diabetic rats, and TE suppressed both of them, whereas TE showed no effect on the polyol pathway metabolites. The results indicate that transition metals play a significant role in the formation of MG and 3-DG via oxidative stress and SSAO activity.
Asunto(s)
Quelantes/uso terapéutico , Cobre/metabolismo , Diabetes Mellitus Experimental/tratamiento farmacológico , Cristalino/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Trientina/uso terapéutico , 8-Hidroxi-2'-Desoxicoguanosina , Amina Oxidasa (conteniendo Cobre)/efectos de los fármacos , Amina Oxidasa (conteniendo Cobre)/metabolismo , Animales , Desoxiglucosa/análogos & derivados , Desoxiglucosa/metabolismo , Desoxiguanosina/análogos & derivados , Desoxiguanosina/metabolismo , Diabetes Mellitus Experimental/metabolismo , Productos Finales de Glicación Avanzada/metabolismo , Cristalino/metabolismo , Masculino , Ornitina/análogos & derivados , Ornitina/metabolismo , Polímeros/metabolismo , Pirimidinas/metabolismo , Piruvaldehído/metabolismo , Ratas , Ratas Sprague-DawleyRESUMEN
Polyamines have been demonstrated to play an important role in adventitious root formation and development in plants. Here, we present a detailed analysis of influence of exogenously added polyamines on adventitious root development and its relationship to cold tolerance in Virginia pine (Pinus virginia Mill.). Our results demonstrated that polyamines putrescine (Put), spermidine (Spd), and spermine (Spm) at 0.001 mM improve rooting frequency and promote root elongation. Put, Spd, and Spm at 0.01-1 mM decrease rooting frequency and reduce root elongation root elongation. Measurements of diamine oxidase (DAO, EC 1.4.3.6) and polyamine oxidase (PAO, EC 1.4.3.4) activities showed that higher DAO and PAO enzyme activities were obtained when high concentrations of polyamines were applied and when plantlets were treated for 5-7 week at 4 degrees C and 16 degrees C. Survival rate of plantlets increased with the treatment of polyamines at low temperature. Polyamines increased mitotic index of cells in root tips of regenerated plantlet cultured on medium containing 0.001 microM Put, Spd, or Spm, but did not increase mitotic index in tissues of needle tips of the same plantlets. These results demonstrated that polyamines promote root elongation and growth by increasing root cell division in regenerated Virginia pine plantlets.
Asunto(s)
Poliaminas Biogénicas/metabolismo , Mitosis/fisiología , Pinus/crecimiento & desarrollo , Raíces de Plantas/crecimiento & desarrollo , Regeneración/fisiología , Amina Oxidasa (conteniendo Cobre)/efectos de los fármacos , Amina Oxidasa (conteniendo Cobre)/metabolismo , Poliaminas Biogénicas/farmacología , Células Cultivadas , Relación Dosis-Respuesta a Droga , Mitosis/efectos de los fármacos , Índice Mitótico , Oxidorreductasas actuantes sobre Donantes de Grupo CH-NH/efectos de los fármacos , Oxidorreductasas actuantes sobre Donantes de Grupo CH-NH/metabolismo , Pinus/efectos de los fármacos , Pinus/metabolismo , Raíces de Plantas/efectos de los fármacos , Raíces de Plantas/metabolismo , Putrescina/metabolismo , Putrescina/farmacología , Regeneración/efectos de los fármacos , Espermidina/metabolismo , Espermidina/farmacología , Espermina/metabolismo , Espermina/farmacología , Tasa de Supervivencia , Regulación hacia Arriba/efectos de los fármacos , Regulación hacia Arriba/fisiología , Poliamino OxidasaRESUMEN
We previously reported that substrates of semicarbazide-sensitive amine oxidase in combination with low concentrations of vanadate exert potent insulin-like effects. Here we performed homology modeling of the catalytic domain of mouse SSAO/VAP-1 and searched through chemical databases to identify novel SSAO substrates. The modeling of the catalytic domain revealed that aromatic residues Tyr384, Phe389, and Tyr394 define a pocket of stable size that may participate in the binding of apolar substrates. We identified a number of amines as substrates of human, rat, and mouse SSAO. The compounds PD0119035, 2,3-dimethoxy-benzylamine, and C-naphthalen-1-yl-methylamine showed high affinity as substrates of rat SSAO. C-Naphthalen-1-yl-methylamine was the only substrate that showed high affinity for human SSAO. C-Naphthalen-1-yl-methylamine and 4-aminomethyl-benzenesulfonamide showed the highest capacity to stimulate glucose transport in isolated rat adipocytes. The impact of these findings on the development of new treatments for diabetes is discussed.
Asunto(s)
Amina Oxidasa (conteniendo Cobre)/química , Amina Oxidasa (conteniendo Cobre)/metabolismo , Moléculas de Adhesión Celular/química , Moléculas de Adhesión Celular/metabolismo , Insulina/metabolismo , Adipocitos/efectos de los fármacos , Adipocitos/metabolismo , Amina Oxidasa (conteniendo Cobre)/efectos de los fármacos , Aminas/metabolismo , Secuencia de Aminoácidos , Animales , Bencilaminas/metabolismo , Bencilaminas/farmacología , Sitios de Unión , Transporte Biológico , Dominio Catalítico , Moléculas de Adhesión Celular/efectos de los fármacos , Células Cultivadas , Evaluación Preclínica de Medicamentos/métodos , Glucosa/metabolismo , Humanos , Insulina/farmacología , Masculino , Metilaminas/química , Metilaminas/metabolismo , Metilaminas/farmacología , Ratones , Modelos Moleculares , Datos de Secuencia Molecular , Naftalenos/química , Naftalenos/metabolismo , Naftalenos/farmacología , Conformación Proteica , Ratas , Ratas Wistar , Homología de Secuencia de Aminoácido , Especificidad por Sustrato , Vanadatos/metabolismo , Vanadatos/farmacologíaRESUMEN
We have recently reported in vivo disruption of collagen and elastin architecture within blood vessel walls resulting from the selective inhibition of the enzyme semicarbazide-sensitive amine oxidase (SSAO). This study further investigates the effects of SSAO inhibition on extracellular matrix deposition by smooth-muscle cells (SMCs) cultured from neonatal rat hearts. SMCs were characterized, SSAO activity was measured, and soluble and insoluble collagen and elastin in the extracellular matrix (ECM) were quantified. Cultured neonatal rat heart SMC exhibited a monotypic synthetic phenotype that likely represents a myofibroblast. Detectable levels of SSAO activity present throughout 30-d culture peaked at 7-14 d, coinciding with the production of ECM. The addition of enzyme inhibitors and alternate SSAO substrates (benzylamine) produced varied and, in some cases, marked changes in SSAO activity as well as in the composition of mature and soluble matrix components. Similar to our previous in vivo findings, in vitro SSAO inhibition produced aberrations in collagen and elastin deposition by heart SMC. Because changes in SSAO activity are associated with cardiovascular pathologic states, this enzyme may play a protective or modulating role by regulating ECM production during pathologic insult.
Asunto(s)
Alilamina/análogos & derivados , Amina Oxidasa (conteniendo Cobre)/metabolismo , Matriz Extracelular/metabolismo , Miocitos del Músculo Liso/metabolismo , Compuestos Alílicos/administración & dosificación , Compuestos Alílicos/antagonistas & inhibidores , Alilamina/administración & dosificación , Alilamina/antagonistas & inhibidores , Amina Oxidasa (conteniendo Cobre)/antagonistas & inhibidores , Amina Oxidasa (conteniendo Cobre)/efectos de los fármacos , Animales , Células Cultivadas , Colágeno/efectos de los fármacos , Colágeno/metabolismo , Relación Dosis-Respuesta a Droga , Elastina/efectos de los fármacos , Elastina/metabolismo , Inhibidores Enzimáticos/administración & dosificación , Matriz Extracelular/efectos de los fármacos , Modelos Animales , Modelos Cardiovasculares , Inhibidores de la Monoaminooxidasa/administración & dosificación , Miocardio/citología , Miocardio/metabolismo , Miocitos del Músculo Liso/clasificación , Miocitos del Músculo Liso/efectos de los fármacos , Propilaminas/administración & dosificación , Propilaminas/antagonistas & inhibidores , Ratas , Ratas Sprague-Dawley , Factores de TiempoRESUMEN
The specific activity and kinetic behaviour of semicarbazide-sensitive amine oxidase (EC 1.4.3.6; SSAO) towards benzylamine, in the rat heart, is affected by in vivo treatment with the non-selective monoamine oxidase (MAO) inhibitor tranylcypromine, but not by the selective MAO-A and -B inhibitors, clorgyline and lazabemide. SSAO activity was increased to 178% of control activity after 7 days of treatment with tranylcypromine. This increase appears to represent an increase in the limiting velocity (V(max)) for benzylamine oxidation with no significant change in the K(m) at that time-point. However, the K(m) for benzylamine oxidation was found to decrease in both controls and treated groups, in a time-dependent manner, during the treatment regime. These findings suggest a link between SSAO and cellular stress and may have importance in the context of the recent finding that tissue-SSAO is identical to a vascular adhesion protein (VAP1), involved in the process of inflammation.
Asunto(s)
Amina Oxidasa (conteniendo Cobre)/metabolismo , Encéfalo/enzimología , Corazón/fisiología , Hígado/enzimología , Inhibidores de la Monoaminooxidasa/farmacología , Monoaminooxidasa/metabolismo , Miocardio/enzimología , Amina Oxidasa (conteniendo Cobre)/efectos de los fármacos , Animales , Encéfalo/efectos de los fármacos , Clorgilina/farmacología , Dopamina/metabolismo , Femenino , Corazón/efectos de los fármacos , Cinética , Hígado/efectos de los fármacos , Monoaminooxidasa/efectos de los fármacos , Ácidos Picolínicos/farmacología , Ratas , Ratas Sprague-Dawley , Serotonina/metabolismo , Fracciones Subcelulares/efectos de los fármacos , Fracciones Subcelulares/metabolismo , Tranilcipromina/farmacologíaRESUMEN
Components of fetal calf serum (FCS) are known to contribute to growth and maintenance of cultured cells. Fetal calf serum supplementation of media also may contribute to the cytotoxicity of other substances to cells grown in vitro. Semicarbazide-sensitive amine oxidase (SSAO) enzyme, present in FCS, metabolizes primary amines and contributes to amine cytotoxicity in vascular smooth muscle cells (VSMC). In cell culture experiments, the media used may greatly affect enzymic activities such as SSAO. In these studies, the SSAO activity in FCS, cultured rat aortic VSMC, and rat plasma was determined in the presence and absence of various culture media. Semicarbazide-sensitive amine oxidase activity in FCS (5-20 microl) was significantly enhanced (approximately 1.5- to 2-fold) in the presence of various culture media, with Dulbecco modified Eagle medium (DMEM), causing the greatest enhancement. Dulbecco modified Eagle medium enhanced the SSAO activity of cultured VSMC in two of the four passages but reduced activity in two passages. Activity in rat plasma was reduced by approximately 25% in the presence of DMEM. The concentrations of various media components, such as glucose, sodium pyruvate, pyridoxine.HCl, and L-glutamine, were not correlated with enhancement. This study identifies an important enhancement effect of culture media on the FCS enzyme, SSAO, although the media components responsible for the enhancement are yet to be identified.
Asunto(s)
Amina Oxidasa (conteniendo Cobre)/metabolismo , Medios de Cultivo , Músculo Liso Vascular/enzimología , Semicarbacidas/farmacología , Amina Oxidasa (conteniendo Cobre)/efectos de los fármacos , Animales , Aorta , Técnicas de Cultivo de Célula/métodos , Supervivencia Celular/efectos de los fármacos , Glucosa/farmacología , Glutamina/farmacología , Masculino , Músculo Liso Vascular/citología , Músculo Liso Vascular/efectos de los fármacos , Ratas , Ratas Sprague-DawleyRESUMEN
We examined whether the antidepressant drugs, such as the dicyclic drug zimeldine, the tricyclic drug imipramine, tetracyclic drug maprotiline, and the non-cyclic drug nomifensine, inhibit in vitro semicarbazide-sensitive amine oxidase (SSAO) activity in monkey brain. The deamination of 1 microM benzylamine was not inhibited at high concentrations of clorgyline or deprenyl, while it was highly sensitive for semicarbazide. When corresponding experiments were performed with 100 microM benzylamine, the opposite results were obtained. The most potent of inhibition of SSAO was observed by imipramine, followed by maprotiline, zimeldine and nomifensine. Inhibition of SSAO was not enhanced by varying the time of preincubation of the enzyme and various antidepressant drugs, indicating direct action on and reversible inhibition of SSAO. We found the tricyclic antidepressant drug to be the most selective inhibitors of SSAO activity in monkey brain, as compared with other type of antidepressant drugs.
Asunto(s)
Amina Oxidasa (conteniendo Cobre)/efectos de los fármacos , Amina Oxidasa (conteniendo Cobre)/metabolismo , Antidepresivos/farmacología , Encéfalo/efectos de los fármacos , Encéfalo/enzimología , Neuronas/efectos de los fármacos , Neuronas/enzimología , Inhibidores de Captación Adrenérgica/farmacología , Animales , Inhibidores de Captación de Dopamina/farmacología , Relación Dosis-Respuesta a Droga , Imipramina/farmacología , Macaca , Masculino , Maprotilina/farmacología , Nomifensina/farmacología , Inhibidores Selectivos de la Recaptación de Serotonina/farmacología , Factores de Tiempo , Zimeldina/farmacologíaRESUMEN
Recently, it has been suggested that ethanol-induced inhibition of liver regeneration results from decreases in hepatic putrescine levels and/or increases in hepatic gamma-aminobutyric acid (GABA)ergic activity. Because putrescine can be metabolized by diamine (DAO) and monoamine (MAO) oxidases to GABA, we documented the effects of acute ethanol exposure on hepatic MAO or DAO activity following partial hepatectomy (PHx) in rats. We also documented the effects of ethanol on GABA transaminase (GABA-T), the enzyme responsible for GABA metabolism in the liver, and tissue putrescine and GABA levels. Adult, male Sprague-Dawley rats (200-250 g) were treated with either ethanol (3 g/kg) or equal volumes of saline by gastric gavage 1 h prior to a 70% PHx or sham surgery. Rats were then sacrificed (n = 5-7/group) at various times (0-72 h) post-PHx. Enzymatic activity and putrescine/GABA levels were determined by standard isotopic techniques and high-performance liquid chromatography respectively. Hepatic DAO activities in ethanol-treated rats were transiently higher than in saline-treated controls (30% increases at 6 h, p < 0.05). Hepatic MAO and GABA-T activities in acute ethanol-treated rats were essentially identical to saline-treated controls. Although hepatic putrescine levels were similar in ethanol- and saline-treated rats, hepatic GABA levels were approximately three times higher in ethanol-treated rats at 12 and 24 h post-PHx (p < 0.0001). In conclusion, the results of this study indicate that acute ethanol exposure has a limited effect on the enzymatic conversion of putrescine to GABA following partial hepatectomy in the liver. The results also indicate that increased GABAergic inhibition rather than decreased putrescine stimulation is more likely to play a role in ethanol-induced inhibition of hepatic regeneration.