RESUMEN
AIMS: Annexin A1 (ANXA1) is a soluble cytoplasmic protein, moving to membranes when calcium levels are elevated. ANXA1 has also been shown to move to the nucleus or outside the cells, depending on tyrosine-kinase signalling, thus interfering in cytoskeletal organization and cell differentiation, mostly in inflammatory and neoplastic processes. The aim was to investigate subcellular patterns of immunohistochemical expression of ANXA1 in neoplastic and non-neoplastic samples from patients with laryngeal squamous cell carcinomas (LSCC), to elucidate the role of ANXA1 in laryngeal carcinogenesis. METHODS AND RESULTS: Serial analysis of gene expression experiments detected reduced expression of ANXA1 gene in LSCC compared with the corresponding non-neoplastic margins. Quantitative polymerase chain reaction confirmed ANXA1 low expression in 15 LSCC and eight matched normal samples. Thus, we investigated subcellular patterns of immunohistochemical expression of ANXA1 in 241 paraffin-embedded samples from 95 patients with LSCC. The results showed ANXA1 down-regulation in dysplastic, tumourous and metastatic lesions and provided evidence for the progressive migration of ANXA1 from the nucleus towards the membrane during laryngeal tumorigenesis. CONCLUSIONS: ANXA1 dysregulation was observed early in laryngeal carcinogenesis, in intra-epithelial neoplasms; it was not found related to prognostic parameters, such as nodal metastases.
Asunto(s)
Anexina A1/metabolismo , Carcinoma de Células Escamosas/metabolismo , Neoplasias Laríngeas/metabolismo , Adulto , Anciano , Anciano de 80 o más Años , Anexina A1/análisis , Anexina A1/genética , Western Blotting , Carcinoma de Células Escamosas/patología , Femenino , Expresión Génica , Humanos , Inmunohistoquímica , Neoplasias Laríngeas/patología , Masculino , Persona de Mediana EdadRESUMEN
Myocardial reperfusion injury is associated with the infiltration of blood-borne polymorphonuclear leukocytes. We have previous described the protection afforded by annexin 1 (ANXA1) in an experimental model of rat myocardial ischemia-reperfusion (IR) injury. We examined the 1) amino acid region of ANXA1 that retained the protective effect in a model of rat heart IR; 2) changes in endogenous ANXA1 in relation to the IR induced damage and after pharmacological modulation; and 3) potential involvement of the formyl peptide receptor (FPR) in the protective action displayed by ANXA1 peptides. Administration of peptide Ac2-26 at 0, 30, and 60 min postreperfusion produced a significant protection against IR injury, and this was associated with reduced myeloperoxidase activity and IL-1beta levels in the infarcted heart. Western blotting and electron microscopy analyses showed that IR heart had increased ANXA1 expression in the injured tissue, associated mainly with the infiltrated leukocytes. Finally, an antagonist to the FPR receptor selectively inhibited the protective action of peptide ANXA1 and its derived peptides against IR injury. Altogether, these data provide further insight into the protective effect of ANXA1 and its mimetics and a rationale for a clinical use for drugs developed from this line of research.
Asunto(s)
Anexina A1/farmacología , Daño por Reperfusión Miocárdica/prevención & control , Péptidos/farmacología , Receptores Inmunológicos/fisiología , Receptores de Péptidos/fisiología , Animales , Anexina A1/química , Anexina A1/metabolismo , Quimiotaxis de Leucocito , Hemodinámica/efectos de los fármacos , Interleucina-1/metabolismo , Cinética , Daño por Reperfusión Miocárdica/metabolismo , Daño por Reperfusión Miocárdica/patología , Miocardio/metabolismo , Neutrófilos/inmunología , Oligopéptidos/farmacología , Peroxidasa/metabolismo , Ratas , Receptores de Formil Péptido , Receptores Inmunológicos/antagonistas & inhibidores , Receptores de Péptidos/antagonistas & inhibidoresRESUMEN
We performed immunogold labeling with an ST-1 monoclonal antibody (IgM), specific for intact heparin, to define the subcellular localization of heparin in mast cells. Rat peritoneal mast cells were fixed by a modified Karnovsky method and embedded in Araldite. Ultrathin sections were first treated with sodium periodate and then sequentially incubated with MAb ST-1, rabbit anti-mouse IgM, and protein A-gold. By transmission electron microscopy, gold particles were localized inside cytoplasmic granules of peritoneal mast cells. In contrast, with the same procedure, no labeling was observed in mast cells from rat intestinal mucosa. Control sections of rat peritoneal or intestinal mucosa mast cells treated with an irrelevant MAb (IgM) did not show any labeling. Treatment with nitrous acid abolished the reactivity of MAb ST-1 with peritoneal mast cells. These results show that different mast cells can be identified regarding their heparin content by immunochemical procedures using MAb ST-1.
Asunto(s)
Anticuerpos Monoclonales , Gránulos Citoplasmáticos/química , Heparina/química , Mastocitos/química , Animales , Heparina/inmunología , Mucosa Intestinal/citología , Masculino , Cavidad Peritoneal/citología , Conejos , Ratas , Ratas WistarRESUMEN
Administration of ovalbumin by aerosol to sensitised rats produced a rapid (15 min) protein exudation in different airway tissues, as determined by Evans blue staining. This was associated with marked mast cell degranulation determined by histological examination, with there being no difference between mucosal and connective tissue mast cells. A 5-day administration regimen with compound 48/80 selectively depleted connective tissue mast cell (positive to berberine staining) without modifying ovalbumin-induced plasma protein extravasation. Treatment of rats with dexamethasone (1 mg/kg, -12 h) or nor-dihydroguaiaretic acid (30 mg/kg i.p., -30 min) significantly reduced ovalbumin-induced protein extravasation and preserved mucosal mast cell morphology. Indomethacin (4 mg/kg i.v., -30 min) exerted no effect on either parameter. In conclusion, we propose the mucosal mast cell as a target cell responsible at least partly for the inhibitory actions of known anti-inflammatory drugs. We suggest an involvement of endogenous leukotriene(s), but not prostanoid(s), in mucosal mast cell activation/degranulation.
Asunto(s)
Inflamación/prevención & control , Mastocitos/efectos de los fármacos , Hipersensibilidad Respiratoria/prevención & control , Administración por Inhalación , Animales , Antiinflamatorios/farmacología , Antígenos/administración & dosificación , Permeabilidad Capilar/efectos de los fármacos , Inhibidores de la Ciclooxigenasa/farmacología , Dexametasona/farmacología , Indometacina/farmacología , Inflamación/inducido químicamente , Inflamación/patología , Inhibidores de la Lipooxigenasa/farmacología , Masculino , Masoprocol/farmacología , Mastocitos/patología , Ovalbúmina/administración & dosificación , Ovalbúmina/inmunología , Ratas , Ratas Wistar , Hipersensibilidad Respiratoria/inducido químicamente , Hipersensibilidad Respiratoria/patología , Tráquea/efectos de los fármacos , Tráquea/patología , p-Metoxi-N-metilfenetilamina/farmacologíaRESUMEN
OBJECTIVE AND DESIGN: We have previously reported a role for annexin-A1 in liver proliferation and tumorogenicity as well as its action as an acute phase protein in a model of endotoxemia in interleukin-6 null mice. MATERIAL AND METHODS: In this study, we have investigated the analysis of the gene and protein expression in annexin-A1 null mice and the wild type livers during foetal and adult life, and in the presence of a proinflammatory stimulus. RESULTS: The data indicate a link between the expression of the annexin-A1 as serine-phosphorylated-protein during early events of the inflammatory response and as tyrosine-phosphorylated-form at later time-points, during the resolution of inflammation. CONCLUSIONS: The study of annexin-A1 post-translation modification may promote a new annexin-A1 peptide discovery programme to treat specific pathologies.
Asunto(s)
Anexina A1/genética , Endotoxemia/inducido químicamente , Regulación de la Expresión Génica , Hígado/fisiología , Procesamiento Proteico-Postraduccional , Animales , Anexina A1/metabolismo , Interleucina-6/genética , Interleucina-6/metabolismo , Lipopolisacáridos/farmacología , Hígado/citología , Hígado/efectos de los fármacos , Hígado/patología , Masculino , Ratones , Ratones Noqueados , Regiones Promotoras GenéticasRESUMEN
OBJECTIVE: We have applied here a model of chronic granulomatous inflammation to study the profile of mast cell activation and their expression of annexin-A1 in the nodular lesion. MATERIALS: Granulomatous inflammation was induced by injection of croton oil and Freund's complete adjuvant (CO/FCA) into the dorsal air-pouches of mice. Skin tissue samples were collected from control group (24 h time-point; i. e. before disease development) and 7, 14, 21, 28 and 42 days post-CO/FCA treatment. RESULTS: Histopathological analyses revealed an on-going inflammation characterized by an increased number of activated mast cells at sites of the chronic inflammatory reaction in all experimental groups. Immunohistochemical analysis showed skin mast cells highly immunoreactive for annexin-A1, both at an initial (day 7) and a delayed (day 28) phase of the inflammatory reaction. CONCLUSIONS: The observed time-dependent modulation of mast cell activation, during the granulomatous injury, indicates that multiple pathways centred on annexin-A1 might become activated at different stages of this chronic inflammatory response, including the delayed and pro-resolving phase.
Asunto(s)
Anexina A1/metabolismo , Granuloma/inmunología , Inflamación/inmunología , Inflamación/patología , Mastocitos/metabolismo , Animales , Enfermedad Crónica , Aceite de Crotón/farmacología , Fármacos Dermatológicos/farmacología , Femenino , Granuloma/inducido químicamente , Granuloma/patología , Inflamación/inducido químicamente , Mastocitos/citología , Ratones , Piel/anatomía & histología , Piel/metabolismo , Piel/patologíaRESUMEN
OBJECTIVE AND DESIGN: To investigate the effect of galectin-1 (Gal-1) and -3 (Gal-3) on leukocyte migration and analyze the expression of both galectins in inflammatory cells using a model of rat peritonitis. MATERIAL OR SUBJECTS: Sprague-Dawley rats (n = 4 per group). TREATMENT: Peritonitis was induced in animals through intraperitoneal injection of carrageenin (1.5 mg/kg) and rat mesenteries were analyzed at different time points (0, 4, 24 and 48h). For pharmacological treatment, rats received intravenous injection of Gal-1 or -3 (3microg/kg) followed by carrageenin. METHODS: Western blotting and immunoelectron microscopy analysis. Statistical analysis was performed using ANOVA followed by Bonferroni test. RESULTS: Pharmacological treatment with Gal-1, but not Gal-3, inhibited (approximately 50 %) leukocyte recruitment into the peritoneal cavity at 4 h time-point. In this early phase, immunogold staining of mesenteries showed a diminished Gal-3 expression in degranulated mast cells and Gal-1 in transmigrated neutrophils (approximately 20 % reduction compared to intravascular cells). In the later phases (24 and 48h), leukocyte turnover was associated with augmented Gal-1 expression in neutrophils and macrophages and Gal-3 in mast cells and macrophages. CONCLUSIONS: These results point to a balanced expression of cell-associated-Gal-1/Gal-3 and might impact on the development of new therapeutic strategies for inflammatory diseases.
Asunto(s)
Modelos Animales de Enfermedad , Galectina 1/metabolismo , Galectina 3/metabolismo , Regulación de la Expresión Génica , Peritonitis/metabolismo , Animales , Carragenina/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Inflamación/inducido químicamente , Inflamación/metabolismo , Macrófagos/metabolismo , Macrófagos/ultraestructura , Masculino , Microscopía Electrónica de Transmisión , Microscopía Inmunoelectrónica , Neutrófilos/metabolismo , Neutrófilos/ultraestructura , Ratas , Ratas Sprague-DawleyRESUMEN
BACKGROUND: There is renewed interest in the role played by specific counter-regulatory mechanisms to control the inflammatory host response, poorly investigated in human pathology. Here, we monitored the expression of two anti-inflammatory mediators, annexin 1 and galectin-1, and assessed their potential link to glucocorticoids' (GCs) effective control of nasal polyposis (NP). METHODS: Total patterns of mRNA and protein expression were analysed by quantitative real-time PCR (qPCR) and Western blotting analyses, whereas ultrastructural immunocytochemistry was used for spatial localization and quantification of each mediator, focusing on mast cells, eosinophils and epithelial cells. RESULTS: Up-regulation of the annexin 1 gene, and down-regulation of galectin-1 gene, was detected in polypoid tissue compared with nasal mucosa. Patient treatment with betamethasone augmented galectin-1 protein expression in polyps. At the cellular level, control mast cells and eosinophils displayed higher annexin 1 expression, whereas marked galectin-1 immunolabelling was detected in the granule matrix of mast cells. Cells of glandular duct epithelium also displayed expression of both annexin 1 and galectin-1, augmented after treatment. CONCLUSION: Mast cells and epithelial cells appeared to be pivotal cell types involved in the expression of both annexin 1 and galectin-1. It is possible that annexin 1 and galectin-1 could be functionally associated with a specific mechanism in NP and that GC exert at least part of their beneficial effects on the airway mucosa by up-regulating, in a specific cell target fashion, these anti-inflammatory agonists.
Asunto(s)
Anexina A1/análisis , Galectina 1/análisis , Mediadores de Inflamación/análisis , Mucosa Nasal/inmunología , Pólipos Nasales/inmunología , Adulto , Anexina A1/genética , Western Blotting/métodos , Eosinófilos/patología , Femenino , Galectina 1/genética , Expresión Génica , Humanos , Masculino , Mastocitos/patología , Microscopía Electrónica de Transmisión , Mucosa Nasal/patología , Pólipos Nasales/patología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodosRESUMEN
The localization of the glucocorticoid-inducible protein annexin 1 (ANX-1) in leukocytes during the process of experimental inflammation has been studied using immunocytochemistry. ANX- 1 immunoreactivity was detected in extravasated neutrophils and eosinophils as well as in resident tissue mast cells. Following injection of carrageenin, the mesenteric tissue was highly inflamed with large presence of leukocytes (predominantly neutrophils with a small percentage of eosinophils) adherent to post-capillary venules and extravasated in the perivascular tissue. ANX-1 immunoreactivity was detected in the cytosol of neutrophils and eosinophils mainly associated with granules and/or vesicles. A good degree of localization in the endosomes was observed in the neutrophils. In both cell types, some ANX-1 immunoreactivity in the nucleus and in the plasma membrane was also detected. Resident mast cells were also activated. Mast cells were positive for ANX-1, without apparent changes in protein content in relation to their activation status. Degranulated mast cells still presented ANX-1 associated with the granule matrix. In conclusion, this study demonstrated the presence of ANX-1 in leukocytes that play a central role in the host inflammatory response. These are the extravasating polymorphonuclear cells, or the resident mast cells. These data provide morphological support to the notion that endogenous and exogenous ANX-1 are able to modulate the reactivity of these cell types, and more generally, of the experimental inflammatory reaction.
Asunto(s)
Anexina A1/metabolismo , Compartimento Celular/fisiología , Quimiotaxis de Leucocito/fisiología , Inflamación/metabolismo , Membranas Intracelulares/metabolismo , Leucocitos/metabolismo , Orgánulos/metabolismo , Animales , Carragenina/farmacología , Endosomas/inmunología , Endosomas/metabolismo , Endosomas/ultraestructura , Inmunohistoquímica , Inflamación/inmunología , Inflamación/patología , Membranas Intracelulares/inmunología , Membranas Intracelulares/ultraestructura , Leucocitos/inmunología , Leucocitos/ultraestructura , Masculino , Microcirculación/inmunología , Microcirculación/metabolismo , Microcirculación/ultraestructura , Microscopía Electrónica , Neutrófilos/inmunología , Neutrófilos/metabolismo , Neutrófilos/ultraestructura , Orgánulos/inmunología , Orgánulos/ultraestructura , Peritonitis/inmunología , Peritonitis/metabolismo , Peritonitis/patología , Ratas , Ratas Sprague-DawleyRESUMEN
Mast cells are present in the eye of chick embryos from the 14th day onward, displaying metachromatic granules, mainly in the iris anterior surface and pectinate ligament. Ultrastructurally these cells show electron-dense granules and a few thin and short cytoplasmic projections in close contact with fibroblasts. Sometimes these contacts are extensive, with long fibroblast projections partially involving the mast cells. Gap junctions between mast cells and fibroblasts are observed only in the eyes of 16- and 20-day-old embryos. These intercellular specializations are represented by a close apposition of cytoplasmic membranes with an extension up to 300 nm. Gap junctions between mast cells and fibroblasts were not observed previously in vivo or in vitro, although in vitro studies have shown that a number of functionally critical interactions may occur between these cells. Our morphological findings suggest that, in vivo, fibroblasts interact with mast cells and may influence their maturation.
Asunto(s)
Ojo/citología , Uniones Comunicantes/ultraestructura , Mastocitos/citología , Animales , Embrión de Pollo , Ojo/embriología , Fibroblastos/citología , Fibroblastos/ultraestructura , Mastocitos/ultraestructura , Microscopía ElectrónicaRESUMEN
Mast cells in the tongue of the bat (Artibeus lituratus) show a well-developed Golgi area and abundant mitochondria in the granule-free perinuclear cytoplasm. Rough endoplasmic reticulum profiles, free ribosomes, mitochondria, bundles of filaments and a great number of secretory granules are found throughout the remaining cytoplasm. The granules, of various shapes and sizes, are simple containing an electron-dense, homogeneous matrix, coarse particles or cylindrical scrolls, or combinations (cylindrical scrolls with either electron-dense, homogeneous matrix or coarse particle contents). Up to now, scroll-containing granules have been considered to be a unique feature of human mast cells.
Asunto(s)
Quirópteros/anatomía & histología , Gránulos Citoplasmáticos/ultraestructura , Mastocitos/ultraestructura , Animales , Femenino , Humanos , Masculino , Lengua/citologíaRESUMEN
OBJECTIVE AND DESIGN: To determine the expression pattern and distribution of the glucocorticoid-inducible protein annexin 1 (ANXA1) in a murine model of chronic granulomatous inflammation. MATERIALS OR SUBJECTS: TO Mouse. TREATMENT: Chronic granulomatous inflammation was induced by injecting into dorsal sub-cutaneous air-pouches in mice, a mixture of croton oil and Freund's complete adjuvant (CO/FCA). METHODS: Western and northern analysis, corticosterone assay, and immunohistochemistry. Statistical analysis was performed using ANOVA followed by Tukey's pair-wise comparisons or Dunnett's multiple comparisons. RESULTS: ANXA1 protein levels changed significantly throughout the 4-week time course, with an initial peak at day 7 and a later elevation at 28 days. ANXA1 mRNA levels peaked at days 1 and 3, with a significant decline at day 7 followed by an upward trend to day 28. Plasma corticosterone measurements taken throughout the time course revealed an increase from 14 days onward, suggesting that corticosterone does not influence ANXA1 expression during the initial stages of the model. Immunogold staining revealed that ANXA1 expression in the inflamed tissue was mainly in extravasated neutrophils, with intact protein (37 kDa) being predominantly observed on the cell membrane. CONCLUSIONS: The pattern of ANXA1 expression indicates that infiltrated neutrophils are responsible for the majority of ANXA1 present both at early and later stages of this model of granulomatous inflammation.
Asunto(s)
Anexina A1/análisis , Granuloma/metabolismo , Inflamación/metabolismo , Animales , Anexina A1/genética , Enfermedad Crónica , Corticosterona/sangre , Femenino , Inmunohistoquímica , Ratones , Neutrófilos/fisiología , ARN Mensajero/análisis , Factores de TiempoRESUMEN
The presence and localization of the anti-inflammatory protein annexin 1 (also known as lipocortin 1) in perivenular rat mast cells was investigated here. Using the rat mesenteric microvascular bed and a combination of morphologic techniques ranging from immunofluorescence to electron microscopy analyses, we detected the presence of annexin 1 in discrete intracellular sites, both in the nucleus and in the cytoplasm. In resting mast cells, most of the protein pool (approximately 80% of the cytosolic portion) was localized to cytoplasmic granules. In agreement with other cell types, treatment of rats with dexamethasone (0.2 mg/kg, ip) increased annexin 1 expression in mast cells, inducing a remarkable appearance of clusters of protein immunoreactivity. This effect was most likely the result of de novo protein synthesis as determined by an increase in mRNA seen by in situ hybridization. Triggering an ongoing experimental inflammatory response (0.3 mg of carrageenin, ip) increased annexin 1 mRNA and protein levels. In conclusion, we report for the first time the localization of annexin 1 in connective tissue mast cells, and its susceptibility not only to glucocorticoid hormone treatment, but also to an experimental acute inflammatory response.
Asunto(s)
Anexinas/análisis , Dexametasona/farmacología , Hibridación in Situ , Inflamación/metabolismo , Mastocitos/química , Animales , Anexinas/genética , Inmunohistoquímica , Masculino , ARN Mensajero/análisis , Ratas , Ratas Sprague-DawleyRESUMEN
Many factors can lead cells to apoptosis during the various stages of cell life. This study was undertaken to characterize germ cell death in the epididymis of the adult Artibeus lituratus by histochemical and immunohistochemical techniques using light microscopy and transmission electron microscopy. The results showed that cells with a nuclear phenotype and ultrastructural characteristics of chromatin compaction were common in apoptosis. The Apoptag test confirmed that the suspected cells were apoptotic. It is suggested that immature germ cells, when released from the germinative epithelium, may be directed towards the epididymis instead of being disposed of in the testicle. Furthermore, intact immature cells can leave the testicle in the initial phases of apoptosis and complete this phenomenon in the epididymis.
Asunto(s)
Apoptosis , Epidídimo/patología , Espermatozoides/patología , Animales , Quirópteros , Masculino , Microscopía Electrónica , Espermatozoides/ultraestructuraRESUMEN
Annexin 1 (ANX-A1) exerts antimigratory actions in several models of acute and chronic inflammation. This is related to its ability to mimic the effect of endogenous ANX-A1 that is externalized on neutrophil adhesion to the postcapillary endothelium. In the present study we monitored ANX-A1 expression and localization in intravascular and emigrated neutrophils, using a classical model of rat peritonitis. For this purpose, a pair of antibodies raised against the ANX-A1 N-terminus (ie, able to recognize intact ANX-A1) or the whole protein (ie, able to interact with all ANX-A1 isoforms) was used by immunofluorescence and immunocytochemistry analyses. The majority ( approximately 50%) of ANX-A1 on the plasma membrane of intravascular neutrophils was intact. Extravasation into the subendothelial matrix caused loss of this pool of intact protein (to approximately 6%), concomitant with an increase in total amount of the protein; only approximately 25% of the total protein was now recognized by the antibody raised against the N-terminus (ie, it was intact). In the cytoplasm of these cells, ANX-A1 was predominantly associated with large vacuoles, possibly endosomes. In situ hybridization confirmed de novo synthesis of ANX-A1 in the extravasated cells. In conclusion, biochemical pathways leading to the externalization, proteolysis, and synthesis of ANX-A1 are activated during the process of neutrophil extravasation.
Asunto(s)
Anexina A1/metabolismo , Endotelio Vascular/patología , Neutrófilos/patología , Animales , Anexina A1/genética , Anexina A1/farmacología , Carragenina/administración & dosificación , Adhesión Celular , Movimiento Celular , Relación Dosis-Respuesta a Droga , Endotelio Vascular/metabolismo , Endotelio Vascular/ultraestructura , Regulación de la Expresión Génica/efectos de los fármacos , Inmunohistoquímica , Hibridación in Situ , Inflamación/genética , Inflamación/metabolismo , Inflamación/patología , Masculino , Mesenterio/efectos de los fármacos , Mesenterio/metabolismo , Mesenterio/patología , Microscopía Electrónica , Neutrófilos/efectos de los fármacos , Peritonitis/inducido químicamente , Peritonitis/metabolismo , Peritonitis/patología , ARN Mensajero/efectos de los fármacos , ARN Mensajero/genética , ARN Mensajero/metabolismo , Ratas , Ratas Sprague-DawleyRESUMEN
We investigated the presence of mast cell granules in macrophages following an in vivo model of an allergic reaction. Injection of ovalbumin (100 microg) into the peritoneal cavity of sensitised mice produced a rapid (within 2 h) influx of neutrophils followed by a slower (after >4 h) eosinophil migration. Ovalbumin treatment induced a high incidence (approximately 50%) of mast cell degranulation compared to control phosphated-buffered saline-treated mice. The majority (approximately 90%) of peritoneal macrophages contained mast cell granules as early as 2 h post-ovalbumin, with lower values at later time-points, as determined by staining with Toluidine blue and Berberine sulphate. This was confirmed by electron microscopy which enabled us to identify the complex mast cell granule sub-structural components in macrophage phagosomes. In conclusion, we used histochemical and ultrastructural analyses to show that mast cell granules become internalised with macrophages during the early stages of an experimental allergic reaction.
Asunto(s)
Hipersensibilidad/fisiopatología , Macrófagos Peritoneales/citología , Mastocitos/citología , Cavidad Peritoneal/citología , Animales , Degranulación de la Célula/efectos de los fármacos , Movimiento Celular/efectos de los fármacos , Femenino , Hipersensibilidad/etiología , Macrófagos Peritoneales/fisiología , Macrófagos Peritoneales/ultraestructura , Mastocitos/fisiología , Mastocitos/ultraestructura , Ratones , Ratones Endogámicos BALB C , Microscopía Electrónica , Ovalbúmina/administración & dosificación , Ovalbúmina/inmunología , Vesículas Secretoras/efectos de los fármacos , Vesículas Secretoras/metabolismo , Vesículas Secretoras/ultraestructura , Factores de TiempoRESUMEN
We assessed here the effect of the glucocorticoid-regulated protein lipocortin 1 (LC1) in a model of rat myocardial ischemia reperfusion. Treatment of animals with human recombinant LC1 at the end of a 25-min ischemic period significantly reduced the extent of infarct size in the area at risk as measured 2 h later, with approximately 50% inhibition at the highest dose tested of 50 microg per rat (equivalent to 5.4 nmol/kg). The protective effect of LC1 was abolished by protein denaturation and not mimicked by the structurally related protein annexin V. A combination of electron and light microscopy techniques demonstrated the occurrence of the myocardial damage at the end of the reperfusion period, with loss of fiber organization. LC1 provided a partial and visible protection. The dose-dependent protection afforded by LC1 was paralleled by lower values of myeloperoxidase activity, tumor necrosis factor a, and macrophage inflammatory protein-1a. The functional link between migrated leukocytes and the myocardial damage was confirmed by electron and light microscopy, and a significantly lower number of extravasated leukocytes was counted in the group of rats treated with LC1 (50 microg). In conclusion, we demonstrate for the first time that LC1 reduces the leukocyte-dependent myocardial damage associated with an ischemia-reperfusion procedure.