RESUMEN
The aim of the study was to examine the association of arachidonic acid-related signal transduction with cerebral metabolism in patients with schizophrenia who have violently and dangerously offended while psychotic. Cerebral 31-phosphorus magnetic resonance spectroscopy was carried out in 11 male patients with schizophrenia who had violently offended (homicide, attempted murder, or wounding with intent to cause grievous bodily harm) while psychotic. Spectra were obtained from 70 x 70 x 70 mm(3) voxels using an image-selected in vivo spectroscopy pulse sequence. Niacin flush testing results were quantified as the volumetric niacin response. There was a strong, and negative, correlation between the volumetric niacin response and the metabolite concentration of inorganic phosphate expressed as a ratio of the total 31-phosphorus signal (p<0.005). Our results suggest that patients with schizophrenia who have violently offended and have poor phospholipid-related signal transduction may have higher levels of cerebral energy metabolism.
Asunto(s)
Corteza Cerebral/química , Niacina/metabolismo , Fosfatos/metabolismo , Esquizofrenia/metabolismo , Violencia , Adulto , Corteza Cerebral/metabolismo , Rubor , Humanos , Imagen por Resonancia Magnética , Masculino , Fosfatos/química , Pruebas Cutáneas , Estadística como AsuntoRESUMEN
Research findings are increasingly reporting evidence of physiological abnormalities in dyslexia and sites for dyslexia have been identified on three chromosomes. It has been suggested that genetic inheritance may cause phospholipid abnormalities in dyslexia somewhat similar to those found in schizophrenia. A key enzyme in phospholipid metabolism, Type IV, or cytosolic, phospholipase A2 (cPLA2), releases arachidonic acid (AA), a 20-carbon fatty acid, which is the major source of production of prostaglandins and leukotrienes. An entirely new assay, which for the first time has enabled determination of the amount of the enzyme rather than its activity, was used to measure cPLA2 in dyslexic-type adults and controls and the two groups were found to differ significantly, the dyslexic-types having more of the enzyme. A report elsewhere of schizophrenics having even greater amounts of the enzyme suggests that dyslexia may be on a continuum with schizophrenia, as may be other neurodevelopmental disorders - which have also been described as phospholipid spectrum disorders.
Asunto(s)
Dislexia/enzimología , Fosfolipasas A/sangre , Adulto , Citosol/enzimología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Fosfolipasas A2 , Esquizofrenia/enzimologíaRESUMEN
Disorders of neurodevelopment include attention deficit hyperactivity disorder, dyspraxia, dyslexia and autism. There is considerable co-morbidity of these disorders and their identification often presents difficulties to those making a diagnosis. This is especially difficult when a multidisciplinary approach is not adopted. All of these disorders have been reported as associated with fatty acid abnormalities ranging from genetic abnormalities in the enzymes involved in phospholipid metabolism to symptoms reportedly improved following dietary supplementation with long chain fatty acids. If definitive disorders of lipid metabolism could be defined then the diagnosis and subsequent management of neurodevelopmental disorders might be transformed. In the identification of those disorders of development which involve lipid metabolism, there are now several tests, measures of lipid metabolism, which could be useful.
Asunto(s)
Apraxias/diagnóstico , Apraxias/metabolismo , Ácidos Grasos/metabolismo , Trastornos Mentales/diagnóstico , Trastornos Mentales/metabolismo , Trastorno por Déficit de Atención con Hiperactividad/diagnóstico , Trastorno por Déficit de Atención con Hiperactividad/metabolismo , Trastorno Autístico/diagnóstico , Trastorno Autístico/metabolismo , Biomarcadores , Pruebas Respiratorias , Niño , Suplementos Dietéticos , Dislexia/diagnóstico , Dislexia/metabolismo , Ácidos Grasos/sangre , Humanos , Errores Innatos del Metabolismo/diagnóstico , Errores Innatos del Metabolismo/metabolismo , Niacina , Fosfolipasas A/metabolismo , Esquizofrenia/diagnóstico , Esquizofrenia/metabolismoRESUMEN
The aim of this pilot study was to evaluate a potential skin test for schizophrenia based on the effect of aqueous methyl nicotinate (AMN) on the production of prostaglandin D2 (PGD2) from skin macrophages and the resultant cutaneous capillary vasodilatation. Four concentrations of AMN were applied topically to the forearm skin in patients and controls, and any resulting vasodilatation was rated as redness after 5 min. The test was carried out on 38 patients with schizophrenia diagnosed according to DSM-III-R criteria, and 22 normal control subjects. At all concentrations of AMN, the schizophrenics were highly significantly different from the controls. One concentration gave the greatest degree of differentiation: at this concentration at 5 min, 83% of schizophrenics but only 23% of controls had a zero or minimal response to AMN. The skin flushing seen after oral administration of nicotinic acid is due to the same reaction, and this has been normal in those with affective illness and neurosis; cyclo-oxygenase inhibitors, e.g., aspirin, give a false-positive result (failure of vasodilatation). This result is consistent with the concept of reduced membrane arachidonic acid levels in schizophrenia. This test may contribute to the reliable diagnosis of schizophrenia.
Asunto(s)
Rubor/inducido químicamente , Ácidos Nicotínicos , Esquizofrenia/diagnóstico , Adulto , Ácido Araquidónico/sangre , Ácidos Docosahexaenoicos/sangre , Relación Dosis-Respuesta a Droga , Membrana Eritrocítica/metabolismo , Estudios de Factibilidad , Femenino , Rubor/fisiopatología , Humanos , Masculino , Persona de Mediana Edad , Proyectos Piloto , Prostaglandina D2/metabolismo , Escalas de Valoración Psiquiátrica , Valores de Referencia , Esquizofrenia/fisiopatología , Sensibilidad y Especificidad , Piel/irrigación sanguínea , Vasodilatación/efectos de los fármacos , Vasodilatación/fisiologíaRESUMEN
Several endogenous peptides, including bradykinin and substance P, have potent inflammatory effects in the joint. Levels of these peptides are regulated by plasma and cell-associated peptide degrading enzymes. One of these peptidases, neutral endopeptidase-24.11 (NEP-24.11), is expressed constitutively and in high density on human synovial cells and is presumed to play a critical role in local regulation of peptide levels in the joint. We examined the role of endogenous NEP-24.11 in regulating bradykinin-mediated effects in an articular model, and investigated the ability of soluble, recombinant human NEP-24.11 to augment the effects of the endogenous enzyme. Our studies demonstrate that endogenous synovial NEP-24.11 does not significantly modulate inflammatory peptide effects on cells when competing with colocalizing peptide receptors expressed in high density. Administration of excess, soluble recombinant NEP-24.11 can overcome this problem, however. Furthermore, the activity of the recombinant enzyme was not compromised in the presence of oxidants or inflammatory joint fluids. Recombinant NEP-24.11 holds promise as a novel therapeutic strategy for the treatment of inflammatory arthritis.
Asunto(s)
Antiinflamatorios no Esteroideos/farmacología , Artritis Reumatoide/enzimología , Neprilisina/metabolismo , Neprilisina/farmacología , Osteoartritis/enzimología , Artritis Reumatoide/metabolismo , Bradiquinina/metabolismo , Calcio/metabolismo , Células Cultivadas , Fibroblastos , Humanos , Neprilisina/antagonistas & inhibidores , Osteoartritis/metabolismo , Oxidantes/farmacología , Prostaglandinas E/metabolismo , Proteínas Recombinantes/farmacología , Superóxidos/metabolismo , Líquido Sinovial/enzimologíaRESUMEN
A recent study determined that cultured human skeletal muscle adult myoblasts, myotubes, and fibroblasts degraded angiotensins and kinins via neutral endopeptidase-24.11 (NEP-24.11: EC 3.4.24.11) and aminopeptidase N (APN: EC 3.4.11.2). Due to the possible importance of other peptides to skeletal muscle blood flow and function, the present study looked specifically at the metabolism of the neurokinins substance P (SP) and neurokinin A (NKA) by skeletal muscle peptidases. The results show that SP is degraded not only by NEP-24.11, but also sequentially by dipeptidyl(amino)peptidase IV (DAP IV: EC 3.4.14.5)/APN. NKA is unaffected by DAP IV but is metabolized by NEP-24.11 and APN. NEP-24.11 was inhibited by phosphoramidon (IC50 = 80 nM), thiorphan and ZINCOV, DAP IV by diprotin A (IC50 = 8 microM), and APN by amastatin (IC50 = 50 nM) and bestatin (IC50 = 100 microM). Skeletal muscle myocyte and fibroblast metabolism of SP and NKA may regulate local skeletal muscle vascular and extravascular functions including SP- and NKA-mediated nerve-induced vasodilation. Inhibition of both NEP-24.11 and DAP IV/APN may increase skeletal muscle blood flow and decrease peripheral vascular resistance via potentiation of local neurokinin levels.
Asunto(s)
Músculo Esquelético/metabolismo , Neuroquinina A/metabolismo , Sustancia P/metabolismo , Adulto , Velocidad del Flujo Sanguíneo/fisiología , Células Cultivadas , Dipeptidil Peptidasa 4/metabolismo , Fibroblastos/metabolismo , Humanos , Músculo Esquelético/irrigación sanguínea , Músculo Esquelético/citología , Neprilisina/metabolismo , Resistencia Vascular/fisiologíaRESUMEN
Inhalation of nail dust is a chronic problem facing podiatric physicians. This problem is compounded in physicians who develop allergic reactions to nail dust. This article defines atopy, reviews the characteristics of nail dust, and suggests methods to reduce the potential hazard that nail dust presents to the podiatric physicians and their employees.
Asunto(s)
Contaminantes Ocupacionales del Aire/efectos adversos , Polvo/efectos adversos , Uñas , Enfermedades Profesionales/etiología , Podiatría , Hipersensibilidad Respiratoria/etiología , Contaminación del Aire/prevención & control , Polvo/prevención & control , Humanos , Enfermedades Profesionales/inmunología , Enfermedades Profesionales/prevención & control , Onicomicosis/complicaciones , Hipersensibilidad Respiratoria/inmunología , Hipersensibilidad Respiratoria/prevención & controlRESUMEN
The most common forms of periungual lesions, both benign and malignant, have been reviewed. The potential psychologic response of patients to these lesions has been discussed. The case reviewed demonstrates how a patient can become overly concerned with thoughts of malignant involvement in these type of lesions. The podiatric physician must be aware of the potential for patients to become anxious and irrational when dealing with periungual lesions.
Asunto(s)
Enfermedades de la Uña/psicología , Trastornos Psicóticos/diagnóstico , Actitud Frente a la Salud , Femenino , Dermatosis del Pie/diagnóstico , Dermatosis del Pie/psicología , Humanos , Persona de Mediana Edad , Enfermedades de la Uña/diagnóstico , Neoplasias/diagnóstico , Neoplasias/psicología , Relaciones Médico-PacienteRESUMEN
Angiotensin (ANG) and kinin metabolizing enzymes, angiotensin-converting enzyme (ACE; EC 3.4.15.1), neutral endopeptidase-24.11 (NEP-24.11; EC 3.4.24.11), and aminopeptidase M (AmM; EC 3.4.11.2), have recently been identified in a purified skeletal muscle glycoprotein fraction. We have analyzed the cellular localization of these enzymes. In cultured human skeletal muscle adult myoblasts, myotubes, and fibroblasts, kinins and angiotensins were metabolized by NEP-24.11 and AmM but not by ACE. NEP-24.11 degraded ANG II, ANG III. and bradykinin (BK) and converted ANG I to the active metabolite ANG(1-7). ANG III was converted to the novel ANG IV metabolite [des-Arg1]ANG III by AmM. These data suggest that, due to their abundance in the body, skeletal muscle myocytes and fibroblasts may play a major role in modulation of the systemic and local effects of angiotensins and kinins. This role could be particularly important in individuals receiving treatment with ACE inhibitors.
Asunto(s)
Angiotensinas/metabolismo , Bradiquinina/metabolismo , Endopeptidasas/metabolismo , Fibroblastos/metabolismo , Músculo Esquelético/metabolismo , Peptidil-Dipeptidasa A/metabolismo , Aminopeptidasas/metabolismo , Angiotensina I/metabolismo , Angiotensina II/metabolismo , Angiotensina III/metabolismo , Células Cultivadas , Humanos , Metionil Aminopeptidasas , Neprilisina/metabolismoRESUMEN
We have identified angiotensin-converting enzyme, neutral endopeptidase-24.11, and aminopeptidase M in a purified glycoprotein fraction of rabbit skeletal muscle membranes. The identification was based on substrate specificity and sensitivity to selective inhibitors. Angiotensin I metabolism was due to angiotensin-converting enzyme-mediated conversion to angiotensin II and neutral endopeptidase-24.11-mediated conversion to angiotensin(1-7). Bradykinin was degraded by angiotensin-converting enzyme and neutral endopeptidase-24.11; angiotensin II by neutral endopeptidase-24.11; and angiotensin III by neutral endopeptidase-24.11 and aminopeptidase M. Thus, the effects of angiotensins and kinins on skeletal muscle blood flow and metabolism may be regulated by local angiotensin-converting enzyme, neutral endopeptidase-24.11, and aminopeptidase M.
Asunto(s)
Angiotensinas/metabolismo , Bradiquinina/metabolismo , Músculo Esquelético/metabolismo , Péptido Hidrolasas/metabolismo , Aminopeptidasas/metabolismo , Angiotensina I/metabolismo , Angiotensina II/metabolismo , Angiotensina III/metabolismo , Animales , Técnicas In Vitro , Metionil Aminopeptidasas , Neprilisina/metabolismo , Peptidil-Dipeptidasa A/metabolismo , ConejosRESUMEN
Respiratory epithelial cell surface neutral endopeptidase 24.11 (NEP-24.11) degrades proinflammatory peptides, and it has been suggested that glucocorticoids may reduce airway inflammation, in part, by upregulation of NEP-24.11. Despite the potential importance of the epithelium as a metabolic barrier, little is known regarding what other peptidases may be present on the epithelial cell surface. Using an immortalized bronchial epithelial cell line (BEAS-2B), we have shown that human epithelial cells express no detectable angiotensin-converting enzyme, carboxypeptidase N, or dipeptidyl(amino)peptidase IV, but express significant levels of aminopeptidase M (AmM), as well as NEP-24.11. The presence of these enzymes was demonstrated via their degradation of biologically active peptides and by flow cytometry. Exposure of cells to the glucocorticoid budesonide (10(-7) M) for up to 5 days did not markedly alter the expression of NEP-24.11 or AmM, as assessed by flow cytometry, nor did glucocorticoid treatment modify rates of peptide hydrolysis by NEP-24.11 or AmM. Thus, BEAS-2B cells have both AmM and NEP-24.11 on their surface, and expression of these enzymes is not altered by glucocorticoids.
Asunto(s)
Bronquios/metabolismo , Glucocorticoides/farmacología , Neprilisina/metabolismo , Aminopeptidasas/metabolismo , Bradiquinina/metabolismo , Bronquios/efectos de los fármacos , Budesonida , Antígenos CD13 , Línea Celular Transformada , Encefalinas/metabolismo , Epitelio/metabolismo , Citometría de Flujo , Humanos , Calidina/metabolismo , Pregnenodionas/farmacología , Sustancia P/metabolismo , Especificidad por SustratoRESUMEN
Analysis of SP and NKA metabolism by human vascular endothelium, relative to that in human plasma, identified integrative, multiple pathways for the processing of circulating SP (but not NKA) by angiotensin-converting enzyme (ACE; EC 3.4.15.1), dipeptidyl(amino)peptidase IV (DAP IV; EC 3.4.14.5), and aminopeptidase M (AmM; EC 3.4.11.2). In contrast, SP and NKA, which may diffuse into or be neurally released within the vessel wall, were both metabolized by smooth muscle neutral endopeptidase-24.11 (NEP-24.11; EC 3.4.24.11). Collectively, these studies indicate peptide-specific and site-specific differential processing of SP and NKA by human plasma and vasculature.
Asunto(s)
Endotelio Vascular/metabolismo , Músculo Liso Vascular/metabolismo , Neuroquinina A/metabolismo , Sustancia P/metabolismo , Células Cultivadas , Humanos , Hidrólisis , InmunoelectroforesisRESUMEN
Intracisternal (ic) injection of the neutral endopeptidase-24.11 inhibitor phosphoramidon (1-100 nmol) produced a dose-dependent inhibition of gastric acid secretion in 2-h pylorus-ligated rats. The response resulted from a reduction in acid concentration and volume. Likewise, ic injection of another neutral endopeptidase-24.11 inhibitor Zincov (200 nmol) produced a 63% inhibition in gastric acid output. In contrast, neither intravenous injection of phosphoramidon (100 nmol) nor ic injection of the aminopeptidase inhibitor amastatin (100 nmol) produced any change in gastric acid secretion. The inhibitory effect of ic phosphoramidon (10 nmol) was not reversed by a dose of naloxone sufficient to antagonize the acid inhibitory effects of ic [D-Ala2-D-met5]enkephalinamide (8.5 nmol). Moreover, phosphoramidon-induced inhibition of acid was not reduced by the centrally effective bombesin antagonist N-acetyl-GRP(20-26)-O-CH3 or by reserpine pretreatment at a dose effective to antagonize ic neurotensin-induced inhibition in acid secretion. These results suggest that an endogenous neutral endopeptidase-24.11 sensitive substrate may act in the brain to inhibit gastric acid output by mechanisms independent of CNS opiate, bombesin or neurotensin activity.
Asunto(s)
Ácido Gástrico/metabolismo , Glicopéptidos/farmacología , Ácidos Hidroxámicos/farmacología , Neprilisina/antagonistas & inhibidores , Péptidos , Animales , Antibacterianos/administración & dosificación , Antibacterianos/farmacología , Interacciones Farmacológicas , Hormonas Gastrointestinales/farmacología , Glicopéptidos/administración & dosificación , Ácidos Hidroxámicos/administración & dosificación , Inyecciones Intravenosas , Inyecciones Intraventriculares , Masculino , Naloxona/farmacología , Neprilisina/metabolismo , Oligopéptidos/farmacología , Ratas , Ratas Sprague-Dawley , Reserpina/farmacologíaRESUMEN
Kinins and substance P have been implicated in the pathogenesis of inflammatory arthritis by virtue of their abilities to induce vasodilation, edema, and pain. The relative biological potencies of these peptides in vivo would depend at least in part upon their rates of catabolism in the joint. We hypothesized that human synovial lining cells may regulate intraarticular levels of kinins and neuropeptides via degradation by cell surface-associated peptidases. We exposed intact human synovial fibroblasts to kinins and substance P, in the presence or absence of specific peptidase inhibitors, and measured the amount of intact substrate remaining and degradation product(s) generated over time. Aminopeptidase M (AmM; EC 3.4.11.2), neutral endopeptidase-24.11 (NEP-24.11; EC 3.4.24.11), and dipeptidyl(amino)peptidase IV (DAP IV; EC 3.4.14.5) were identified on the cell surface of synovial cells. Bradykinin degradation was due entirely to NEP-24.11 (1.39 +/- 0.29 nmol/min per well). Lysylbradykinin was also degraded by NEP-24.11 (0.80 +/- 0.19 nmol/min per well); however, in the presence of phosphoramidon, AmM-mediated conversion to bradykinin (3.74 +/- 0.46 nmol/min per well) could be demonstrated. The combined actions of NEP-24.11 (0.93 +/- 0.15 nmol/min per well) and DAP IV (0.84 +/- 0.18 nmol/min per well) were responsible for the degradation of substance P. AmM (2.44 +/- 0.33 nmol/min per well) and NEP-24.11 (1.30 +/- 0.45 nmol/min per well) were responsible for the degradation of the opioid peptide, [Leu5]enkephalin. The identity of each of the three peptidases was confirmed via synthetic substrate hydrolysis, inhibition profile, and immunological identification. The profiles of peptidase enzymes identified in cells derived from rheumatoid and osteoarthritic joints were identical. These data demonstrate the human synovial fibroblast to be a rich source of three specific peptidases and suggest that it may play a prominent role in regulating peptide levels in the joint.
Asunto(s)
Cininas/metabolismo , Neuropéptidos/metabolismo , Membrana Sinovial/metabolismo , Antígenos CD/análisis , Células Cultivadas , Endopeptidasas/análisis , Encefalinas/metabolismo , Fibroblastos/metabolismo , Citometría de Flujo , Humanos , Interleucina-1/farmacología , Sustancia P/metabolismoRESUMEN
Bradykinin (BK) receptor agonists and antagonists contain modifications that confer resistance to specific peptidases. In control studies, rat plasma degraded BK (10.3 +/- 0.3 nmol/min/ml) via angiotensin-converting enzyme (ACE; EC 3.4.15.1; 5.2 +/- 0.3 nmol/min/ml), carboxypeptidase N (CPN; EC 3.4.17.3; 3.2 +/- 0.4 nmol/min/ml), aminopeptidase P (APP; EC 3.4.11.9; 0.6 +/- 0.2 nmol/min/ml), and other (unidentified) activity (2.1 +/- 0.6 nmol/min/ml). In contrast, BK agonist analogs were hydrolyzed more slowly due to selective resistance to these plasma peptidases. In addition to Lys-Lys-BK (B1087), which is partially resistant to ACE, [Hyp3,Phe8-r-Arg9]BK (B7642) was completely resistant to ACE, CPN, and the unidentified plasma activity (1.9 +/- 0.3 nmol/min/ml), and D-Arg0[Hyp3,Phe8-r-Arg9]BK (B7644) was resistant to all plasma hydrolysis, including APP (less than 0.2 nmol/min/ml). In vivo ACE-resistant B1087 exhibited a depressor potency and duration of action greater than BK and equivalent to that of BK in the presence of the ACE inhibitor enalapril. Although the B7642 and B7644 agonists were also more potent and longer acting than BK, the increases were no more than that seen for B1087, despite their additional resistance to CPN (B7642) and CPN and APP (B7644). The duration of action of these analogs was, however, increased after renal ligation. These data demonstrate the importance of ACE to the metabolism of circulating BK and BK analogs. In contrast, resistance to CPN and APP are not associated with further potentiation. Beyond ACE resistance, it is likely that the development of more potent, longer-acting BK agonists and antagonists will relate to other factors, such as renal processing independent of CPN and APP.
Asunto(s)
Aminopeptidasas/fisiología , Presión Sanguínea/efectos de los fármacos , Bradiquinina/metabolismo , Lisina Carboxipeptidasa/fisiología , Peptidil-Dipeptidasa A/fisiología , Receptores de Neurotransmisores/efectos de los fármacos , Animales , Bradiquinina/farmacología , Técnicas In Vitro , Riñón/metabolismo , Masculino , Ratas , Ratas Endogámicas , Receptores de BradiquininaRESUMEN
Recent studies have demonstrated that Fischer-344 rats from Japanese Charles River Inc. specifically lack dipeptidyl(amino)peptidase IV (DAP IV-negative; EC 3.4.14.5), whereas Fischer-344 rats from sources within the United States (DAP IV-positive) possess normal DAP IV activity. In the present study, plasma from DAP IV-positive rats metabolized substance P (SP) (5.37 +/- 0.25 nmol/min/ml) via the actions of angiotensin-converting enzyme (EC 3.4.15.1) (1.86 +/- 0.50 nmol/min/ml) and DAP IV (2.56 +/- 0.42 nmol/min/ml). DAP IV sequentially converted SP to SP[3-11] and SP[5-11]. The SP[5-11] metabolite was then rapidly hydrolyzed by plasma aminopeptidase M (AmM; EC 3.4.11.2) (36.2 +/- 4.2 nmol/min/ml). In contrast, SP metabolism by plasma from DAP IV-negative rats was less than half that of control animals (2.14 +/- 0.06 nmol/min/ml), due to a complete lack of DAP IV hydrolysis. The absence of DAP IV was not associated with any differences in angiotensin-converting enzyme-mediated hydrolysis of SP (1.45 +/- 0.11 nmol/min/ml) or AmM-mediated hydrolysis of SP[5-11] (37.1 +/- 0.9 nmol/min/ml). Consistent with this deficiency in SP metabolism, SP was more potent in vivo in stimulating salivary secretion in DAP IV-negative rats compared to DAP IV-positive animals. Potentiation was specific in that SP[5-11], an SP fragment resistant to DAP IV, was equipotent in DAP IV-negative and positive animals. SP[5-11]-induced salivary secretion was potentiated in both strains when AmM-mediated hydrolysis was inhibited by amastatin (20 nmol/min, i.v.). These data provide direct evidence for a significant role for DAP IV and AmM in the in vivo processing of SP and active SP metabolites.
Asunto(s)
Aminopeptidasas/fisiología , Dipeptidil-Peptidasas y Tripeptidil-Peptidasas/fisiología , Sustancia P/metabolismo , Animales , Antígenos CD13 , Dipeptidil Peptidasa 4 , Femenino , Masculino , Fragmentos de Péptidos/metabolismo , RatasRESUMEN
The present investigation determined that a commercially available aminopeptidase M (AmM, Sigma Chemical) can be utilized to lower blood pressure in normotensive and hypertensive rats. In vitro analyses indicated that the predominant peptidase present in this preparation was AmM; however, it also contained some aminopeptidase A (AmA) and less DAP IV. Although no DAP IV-mediated metabolism of angiotensin II (AII) or angiotensin III (AIII) was measured, both AmM and AmA metabolized AII and AIII. Upon further examination, it appeared that AII could be converted to AIII by either AmM or AmA; however, Arg was cleaved from the N-Terminal of AIII predominantly by AmM. The aminopeptidase inhibitors actinonin (AC), amastatin (AM), and bestatin (BE) effectively blocked the AmM-induced hydrolysis of the Asp-Arg bond of AII, and the Arg-Val bond of AIII. The activity of AmA was inhibited by AM but was relatively resistant to inhibition by AC and BE. Next, exogenous aminopeptidase replacement was employed in the anesthetized spontaneously hypertensive rat (SHR) in an attempt to temporarily correct a hypothesized brain deficiency of receptor-associated peptidases and lower blood pressure. Third-ventricle infusion of AmM produced significant drops in blood pressure and heart rate in both SHRs and Wistar-Kyoto normotensive controls. Pretreatment with AC or BE was particularly effective at interfering with the subsequent AmM-induced hypotensive effect, while AM was less effective. The central mechanisms underlying these effects are in need of further investigation; however, they are at least partially dependent upon the brain angiotensin system.
Asunto(s)
Aminopeptidasas/farmacología , Antihipertensivos/farmacología , Presión Sanguínea/efectos de los fármacos , Ventrículos Cerebrales/fisiología , Aminopeptidasas/administración & dosificación , Aminopeptidasas/metabolismo , Animales , Antihipertensivos/administración & dosificación , Antígenos CD13 , Ventrículos Cerebrales/efectos de los fármacos , Frecuencia Cardíaca/efectos de los fármacos , Inmunoelectroforesis Bidimensional , Infusiones Parenterales , Masculino , Ratas , Ratas Endogámicas SHR , Ratas Endogámicas WKYRESUMEN
In addition to plasma metabolism of substance P (SP) by angiotensin converting enzyme (ACE; EC 3.4.15.1) (less than 1.0 nmol/min/ml), the majority of SP hydrolysis by rat and human plasma was due to dipeptidyl(amino)peptidase IV (DAP IV; EC 3.4.14.5) (3.15-5.91 nmol/min/ml), which sequentially converted SP to SP(3-11) and SP(5-11). In turn, the SP(5-11) metabolite was rapidly hydrolyzed by rat and human plasma aminopeptidase M (AmM; EC 3.4.11.2) (24.2-25.5 nmol/min/ml). The Km values of SP for DAP IV and of SP(5-11) for AmM ranged from 32.7 to 123 microM. In contrast, neurokinin A (NKA) was resistant to both ACE and DAP IV but was subject to N-terminal hydrolysis by AmM (3.76-10.8 nmol/min/ml; Km = 90.7 microM). These data demonstrate differential processing of SP and NKA by specific peptidases in rat and human plasma.
Asunto(s)
Antibacterianos , Neuroquinina A/sangre , Péptidos , Sustancia P/sangre , Aminopeptidasas/sangre , Animales , Antígenos CD13 , Captopril/farmacología , Dipeptidil Peptidasa 4 , Dipeptidil-Peptidasas y Tripeptidil-Peptidasas/sangre , Humanos , Técnicas In Vitro , Cinética , Masculino , Oligopéptidos/farmacología , Fragmentos de Péptidos/sangre , Peptidil-Dipeptidasa A/sangre , Ratas , Especificidad de la EspecieRESUMEN
In addition to angiotensin I converting enzyme (ACE; EC 3.4.15.1) and carboxypeptidase N (CPN; EC 3.4.17.3), other peptidases contribute to bradykinin (BK) degradation in plasma. Rat plasma degraded BK by hydrolysis of the N-terminal Arg1-Pro2 bond, and the characteristics of hydrolysis are consistent with identification of aminopeptidase P (APP; EC 3.4.11.9) as the responsible enzyme. BK and BK[1-5] N-terminal hydrolysis was optimal at neutral pH, was inhibited by 2-mercaptoethanol, dithiothreitol, o-phenanthroline and EDTA, but was unaffected by the aminopeptidase inhibitors amastatin, puromycin and diprotin A, the endopeptidase-24.11 inhibitors phosphoramidon and ZINCOV, and the ACE and CPN inhibitors captopril and D,L-mercapto-methyl-3-guanidinoethylthiopropanoic acid (MERGETPA), respectively. Although kallidin (Lys-BK) was not metabolized directly by APP, conversion to BK by plasma aminopeptidase M (EC 3.4.11.2) resulted in subsequent degradation by APP. BK analogs containing N-terminal Arg1-Pro2 bonds, including [Tyr8-(OMe)] BK and [Phe8 psi(CH2NH)Arg9]BK (B2 agonists), des-Arg9-BK and [D-Phe8]des-Arg9-BK (B1 agonists), and [Leu8]des-Arg9-BK (B1 antagonist), were degraded by APP with Km and Vmax values comparable to those found for BK (Km = 19.7 +/- 2.6 microM; Vmax = 12.1 +/- 1.2 nmol/min/mL). In contrast, B2 antagonists containing D-Arg0 N-termini, including D-Arg[Hyp3,Thi5.8,D-Phe7]BK and D-Arg[Hyp3,D-Phe7,Phe8 psi(CH2NH)Arg9]BK, were resistant to APP-mediated hydrolysis. These data support a role for plasma aminopeptidase P in the degradation of circulating kinins, and a variety of B2 and B1 kinin agonists and antagonists. However, APP does not participate in the degradation of D-Arg0-containing antagonists.