RESUMEN
OBJECTIVE: To analyze soluble and membrane-bound peptidase activities in the tonsils and adenoids removed from patients with adenoid hyperplasia, tonsillar hyperplasia and chronic tonsillitis. METHODS: A total of 48 tissue samples from patients undergoing adenoidectomy and tonsillectomy for adenoid hyperplasia, tonsillar hyperplasia or chronic tonsillitis were analyzed. The catalytic activity of a pool of peptidases in the soluble (dipeptidyl peptidase IV, aminopeptidase A, aminopeptidase N and cystinyl aminopeptidase) and membrane-bound (prolyl endopeptidase, aspartyl aminopeptidase, aminopeptidase B and pyroglutamyl peptidase I) fractions was measured fluorometrically. RESULTS: The activity of membrane-bound aminopeptidase B was higher in cases of chronic tonsillitis and adenoid hyperplasia than in tonsillar hyperplasia, p=0.004. Soluble dipeptidyl peptidase IV and membrane-bound pyroglutamyl peptidase I were found to be more active in tissues from male chronic tonsillitis tissues, p<0.05, while membrane-bound aminopeptidase B activity was higher in tissues of females with tonsillar hyperplasia, p<0.001. In the case of chronic tonsillitis, soluble aminopeptidase A was found to have a higher level of activity in tissues from children than those from adults, p=0.005. CONCLUSIONS: Our results suggest a potential role of soluble aminopeptidase A, soluble dipeptidyl peptidase IV, membrane-bound aminopeptidase B and membrane-bound pyroglutamyl peptidase I in the pathobiology of adenoid hyperplasia, tonsillar hyperplasia and chronic tonsillitis that is differently regulated as a function of gender. These finfings may modify in the future the clinical approach to these diseases.
Asunto(s)
Tonsila Faríngea/metabolismo , Aminopeptidasas/metabolismo , Tonsila Palatina/metabolismo , Tonsila Faríngea/patología , Aminopeptidasas/análisis , Análisis de Varianza , Biomarcadores/análisis , Niño , Preescolar , Enfermedad Crónica , Estudios de Cohortes , Dipeptidil Peptidasa 4/análisis , Dipeptidil Peptidasa 4/metabolismo , Femenino , Glutamil Aminopeptidasa/análisis , Glutamil Aminopeptidasa/metabolismo , Humanos , Hiperplasia/metabolismo , Hiperplasia/patología , Masculino , Tonsila Palatina/patología , Piroglutamil-Peptidasa I/análisis , Piroglutamil-Peptidasa I/metabolismo , Estudios Retrospectivos , Sensibilidad y Especificidad , Índice de Severidad de la Enfermedad , Tonsilectomía/métodos , Tonsilitis/metabolismo , Tonsilitis/patología , Tonsilitis/cirugíaRESUMEN
Acid (aspartyl), basic (arginyl) and neutral (alanyl) aminopeptidases degrade angiotensins, vasopressin, oxytocin, bradykinin and enkephalins. These peptides regulate memory, energy homeostasis, water-salt balance and blood pressure, functions that are mainly exerted in the hippocampus and hypothalamus, and that can be affected by diabetes mellitus. To evaluate the relationship between the diabetes mellitus and processing and inactivation roles of these representative aminopeptidases, we measured their activities in both brain structures of control and streptozotocin-diabetic rats. Hypothalamic soluble aspartyl and arginyl aminopeptidases presented significant decreased activity levels in diabetic rats, which were mitigated by insulin therapy. In addition to membrane-bound puromycin sensitive and insensitive alanyl aminopeptidases, its soluble puromycin sensitive form did not differ between diabetic and control rats in both brain structures. Glucose and/or insulin did not seem to alter in vitro the hypothalamic activities of soluble aspartyl and arginyl aminopeptidases. The implied hypothalamic control of regulatory peptide activity by aspartyl and arginyl aminopeptidases supports the hypothesis that the hydrolytic ability of these enzyme types could be a common link for the disruptions of water-salt balance, blood pressure and energy homeostasis in diabetes mellitus.
Asunto(s)
Aminopeptidasas/metabolismo , Encefalopatías Metabólicas/enzimología , Encefalopatías Metabólicas/etiología , Diabetes Mellitus Experimental/complicaciones , Hipocampo/enzimología , Hipotálamo/enzimología , Aminopeptidasas/análisis , Animales , Presión Sanguínea/efectos de los fármacos , Presión Sanguínea/fisiología , Encefalopatías Metabólicas/fisiopatología , Antígenos CD13/análisis , Antígenos CD13/metabolismo , Membrana Celular/efectos de los fármacos , Membrana Celular/enzimología , Regulación hacia Abajo/efectos de los fármacos , Regulación hacia Abajo/fisiología , Enfermedades del Sistema Endocrino/enzimología , Enfermedades del Sistema Endocrino/etiología , Enfermedades del Sistema Endocrino/fisiopatología , Glutamil Aminopeptidasa/análisis , Glutamil Aminopeptidasa/metabolismo , Hipocampo/fisiopatología , Homeostasis/fisiología , Hipotálamo/fisiopatología , Insulina/metabolismo , Insulina/farmacología , Masculino , Neuronas/efectos de los fármacos , Neuronas/enzimología , Neuropéptidos/metabolismo , Inhibidores de la Síntesis de la Proteína/farmacología , Puromicina/farmacología , Ratas , Ratas Wistar , Equilibrio Hidroelectrolítico/fisiologíaRESUMEN
The aminopeptidase activities of snake venoms from Gloydius blomhoffi brevicaudus, Gloydius halys blomhoffii, Trimeresurus flavoviridis, Bothrops jararaca and Crotalus atrox were investigated. Aminopeptidase A (APA), aminopeptidase B and aminopeptidase N activities were present in all snake venoms. The strongest APA activity was found in venom from G. blomhoffi brevicaudus. The susceptibility to metallopeptidase inhibitors and the pH optimum of the partially purified enzyme from G. blomhoffi brevicaudus venom were similar to those of known APAs from mammals. A G. blomhoffi brevicaudus venom gland cDNA library was screened to isolate cDNA clones using probes based on highly conserved amino acid sequences in known APAs. Molecular cloning of APA from G. blomhoffi brevicaudus venom predicted that it was a type II integral membrane protein containing 958 amino acid residues with 17 potential N-linked glycosylation sites. It possessed a His-Glu-Xaa-Xaa-His-(Xaa)(18)-Glu zinc binding motif that allowed the classification of this protein as a member of the M1 family of zinc-metallopeptidases, or gluzincins. The deduced amino acid sequence shows approximately 60% sequence identity to mammalian APA sequences. This is the first study to report the primary structure of APA from a reptile.
Asunto(s)
Venenos de Crotálidos/enzimología , Glutamil Aminopeptidasa/genética , Proteínas de la Membrana/genética , Filogenia , Viperidae , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Cromatografía en Gel , Clonación Molecular , Análisis por Conglomerados , Cartilla de ADN/genética , ADN Complementario/genética , Glutamil Aminopeptidasa/análisis , Proteínas de la Membrana/análisis , Datos de Secuencia Molecular , Conformación Proteica , Análisis de Secuencia de ADN , Especificidad de la EspecieRESUMEN
Pericytes were isolated and cultured from mouse cerebroparenchymal microvessels. A single pericyte clone was three-dimensionally cultured in a collagen gel by adding tensile stress, resulting in the reconstruction of narrow stringy fibers. When the contractility of these fibers was evaluated isometrically, they contracted in response to acetylcholine (ACh)1 or noradrenaline; this was accompanied by an increase in intracellular calcium concentration ([Ca(2+)]i). The fibers that were pre-contracted by ACh were completely relaxed by papaverine, which is a smooth-muscle relaxant. Moreover, the muscarinic ACh receptor-antagonist atropine depressed the [Ca(2+)]i response that was induced by ACh. This study demonstrates for the first time the quantitative measurement of the contractions produced by cultured microvascular pericytes from mouse brain parenchyma.