Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 10 de 10
Filtrar
1.
Arterioscler Thromb Vasc Biol ; 30(3): 395-402, 2010 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-20139367

RESUMEN

OBJECTIVE: Mouse aorta smooth muscle cells (SMC) express tumor necrosis factor receptor superfamily member 1A (TNFR-1) and lymphotoxin beta-receptor (LTbetaR). Circumstantial evidence has linked the SMC LTbetaR to tertiary lymphoid organogenesis in hyperlipidemic mice. Here, we explored TNFR-1 and LTbetaR signaling in cultured SMC. METHODS AND RESULTS: TNFR-1 signaling activated the classical RelA NF-kappaB pathway, whereas LTbetaR signaling activated the classical RelA and alternative RelB NF-kappaB pathways, and both signaling pathways synergized to enhance p100 inhibitor processing to the p52 subunit of NF-kappaB. Microarrays showed that simultaneous TNFR-1/LTbetaR activation resulted in elevated mRNA encoding leukocyte homeostatic chemokines CCL2, CCL5, CXCL1, and CX3CL1. Importantly, SMC acquired features of lymphoid tissue organizers, which control tertiary lymphoid organogenesis in autoimmune diseases through hyperinduction of CCL7, CCL9, CXCL13, CCL19, CXCL16, vascular cell adhesion molecule-1, and intercellular adhesion molecule-1. TNFR-1/LTbetaR cross-talk resulted in augmented secretion of lymphorganogenic chemokine proteins. Supernatants of TNFR-1/LTbetaR-activated SMC markedly supported migration of splenic T cells, B cells, and macrophages/dendritic cells. Experiments with ltbr(-/-) SMC indicated that LTbetaR-RelB activation was obligatory to generate the lymphoid tissue organizer phenotype. CONCLUSIONS: SMC may participate in the formation of tertiary lymphoid tissue in atherosclerosis by upregulation of lymphorganogenic chemokines involved in T-lymphocyte, B-lymphocyte, and macrophage/dendritic cell attraction.


Asunto(s)
Diferenciación Celular/fisiología , Tejido Linfoide/citología , Receptor beta de Linfotoxina/fisiología , Miocitos del Músculo Liso/citología , FN-kappa B/fisiología , Receptores Tipo I de Factores de Necrosis Tumoral/fisiología , Transducción de Señal/fisiología , Animales , Anticuerpos Monoclonales/farmacología , Aorta/citología , Aorta/efectos de los fármacos , Aorta/fisiología , Aterosclerosis/patología , Aterosclerosis/fisiopatología , Movimiento Celular/fisiología , Células Cultivadas , Modelos Animales de Enfermedad , Tejido Linfoide/fisiología , Receptor beta de Linfotoxina/genética , Receptor beta de Linfotoxina/inmunología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Miocitos del Músculo Liso/efectos de los fármacos , Miocitos del Músculo Liso/fisiología , Factor de Necrosis Tumoral alfa/farmacología
2.
Nat Med ; 10(9): 966-73, 2004 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-15322539

RESUMEN

Activation of the 5-lipoxygenase (5-LO) pathway leads to the biosynthesis of proinflammatory leukotriene lipid mediators. Genetic studies have associated 5-LO and its accessory protein, 5-LO-activating protein, with cardiovascular disease, myocardial infarction and stroke. Here we show that 5-LO-positive macrophages localize to the adventitia of diseased mouse and human arteries in areas of neoangiogenesis and that these cells constitute a main component of aortic aneurysms induced by an atherogenic diet containing cholate in mice deficient in apolipoprotein E. 5-LO deficiency markedly attenuates the formation of these aneurysms and is associated with reduced matrix metalloproteinase-2 activity and diminished plasma macrophage inflammatory protein-1alpha (MIP-1alpha; also called CCL3), but only minimally affects the formation of lipid-rich lesions. The leukotriene LTD(4) strongly stimulates expression of MIP-1alpha in macrophages and MIP-2 (also called CXCL2) in endothelial cells. These data link the 5-LO pathway to hyperlipidemia-dependent inflammation of the arterial wall and to pathogenesis of aortic aneurysms through a potential chemokine intermediary route.


Asunto(s)
Aneurisma de la Aorta Abdominal/etiología , Araquidonato 5-Lipooxigenasa/metabolismo , Regulación de la Expresión Génica , Hiperlipidemias/complicaciones , Leucotrienos/biosíntesis , Macrófagos/metabolismo , Proteínas Activadoras de la 5-Lipooxigenasa , Análisis de Varianza , Animales , Western Blotting , Proteínas Portadoras/metabolismo , Quimiocina CCL2/sangre , Quimiocina CCL3 , Quimiocina CCL4 , Quimiocina CXCL1 , Quimiocinas CXC/metabolismo , Colatos , Tejido Conectivo/metabolismo , Citocinas/sangre , Cartilla de ADN , Dieta Aterogénica , Técnicas Histológicas , Humanos , Inmunohistoquímica , Péptidos y Proteínas de Señalización Intercelular/metabolismo , Leucotrieno D4/metabolismo , Proteínas Inflamatorias de Macrófagos/metabolismo , Metaloproteinasa 2 de la Matriz/metabolismo , Proteínas de la Membrana/metabolismo , Ratones , Ratones Endogámicos C57BL , Neovascularización Patológica/metabolismo , ARN/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
3.
Circ Res ; 100(2): 255-62, 2007 Feb 02.
Artículo en Inglés | MEDLINE | ID: mdl-17170365

RESUMEN

The endothelial cell protein C receptor (EPCR) is expressed on endothelial cells and regulates the protein C anticoagulant pathway via the thrombin-thrombomodulin complex. Independent of its anticoagulant activity, activated protein C (APC) can directly signal to endothelial cells and upregulate antiapoptotic and antiinflammatory genes. Here we show that vascular smooth muscle cells (SMCs) also express EPCR. EPCR protein on SMCs was detected by flow cytometry and Western blotting. EPCR mRNA was identified by quantitative RT-PCR. To examine the functionality of EPCR, intracellular signaling in APC-stimulated SMCs was analyzed by determination of intracellular free calcium transients using confocal laser scanning microscopy. Phosphorylation of extracellular signal-regulated kinases 1 and 2 (ERK-1/2) was detected by immunoblotting. APC-induced ERK-1/2 phosphorylation was inhibited by an anti-EPCR antibody and by a cleavage site blocking anti-PAR-1 antibody, indicating that binding of APC to EPCR and cleavage of protease-activated receptor-1 (PAR-1) were involved. APC elicited an increase in [(3)H]-thymidine incorporation. The mitogenic effect of APC was significantly enhanced in the presence of thrombin. EPCR expression was also detected in SMCs in the fibrous cap of human carotid artery plaques. The present data demonstrate functionally active EPCR in SMCs and suggest that EPCR-bound APC might modulate PAR-1-mediated responses of SMCs to vascular injury.


Asunto(s)
Antígenos CD/biosíntesis , Antígenos CD/genética , Endotelio Vascular/metabolismo , Regulación de la Expresión Génica/fisiología , Músculo Liso Vascular/metabolismo , Miocitos del Músculo Liso/metabolismo , Proteína C/metabolismo , Receptores de Superficie Celular/biosíntesis , Receptores de Superficie Celular/genética , Antígenos CD/fisiología , Estenosis Carotídea/enzimología , Estenosis Carotídea/metabolismo , Estenosis Carotídea/patología , Células Cultivadas , Técnicas de Cocultivo , Receptor de Proteína C Endotelial , Endotelio Vascular/enzimología , Endotelio Vascular/patología , Humanos , Músculo Liso Vascular/enzimología , Músculo Liso Vascular/patología , Miocitos del Músculo Liso/enzimología , Miocitos del Músculo Liso/patología , Unión Proteica/fisiología , Receptores de Superficie Celular/fisiología
4.
Prostaglandins Other Lipid Mediat ; 84(3-4): 108-15, 2007 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-17991613

RESUMEN

The 5-lipoxygenase (5-LO) pathway generates lipid mediators, i.e. the cysteinyl leukotrienes (cysLTs) LTC(4)/LTD(4) and LTB(4). CysLT receptors are expressed in endothelial cells (EC) and EC cysLT(2)-R activation induces diverse pro-inflammatory genes in vitro. We now report that LTD(4) promotes formation of an atherosclerosis-protective and anti-thrombotic eicosanoid by markedly up-regulating EC cyclooxygenase-2 (COX-2). CysLT-induced COX-2 transcripts were transiently up-regulated as determined by microarray and QRT-PCR analyses though COX-2 protein remained elevated for several hours. Prostacyclin formation, measured as its stable metabolite 6-keto-PGF(1alpha), was increased several fold in LTD(4)-stimulated ECs, and was inhibited by the COX-2-specific inhibitor, NS-398. COX-2 up-regulation was Ca(2+)-dependent and was partially blocked by cyclosporin A indicating that the 5-LO/COX-2 cross-talk involved signaling through a nuclear factor of activated T cells (NFAT) dependent pathway. Since prostacyclin is a major blood vessel-protective and anti-thrombotic eicosanoid, the EC cysLT(2)-R may limit its otherwise pro-inflammatory actions through a protective Ca(2+)/calcineurin/NFAT-dependent COX-2 feedback loop.


Asunto(s)
Araquidonato 5-Lipooxigenasa/metabolismo , Ciclooxigenasa 2/metabolismo , Células Endoteliales/metabolismo , Proteínas de la Membrana/metabolismo , Receptor Cross-Talk , Receptores de Leucotrienos/metabolismo , Señalización del Calcio , Ciclooxigenasa 2/genética , Ciclosporina/metabolismo , Células Endoteliales/enzimología , Epoprostenol/biosíntesis , Epoprostenol/metabolismo , Humanos , Leucotrieno D4/metabolismo , ARN Mensajero/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Venas Umbilicales/citología , Regulación hacia Arriba
5.
Biochim Biophys Acta ; 1736(1): 30-7, 2005 Sep 05.
Artículo en Inglés | MEDLINE | ID: mdl-16081317

RESUMEN

Leukotrienes (LTs) are powerful inflammatory lipid mediators derived from the 5-lipoxygenase (5-LO) cascade of arachidonic acid. Recent clinical, population genetic, cell biological, and mouse studies indicate participation of the 5-LO pathway in atherogenesis and arterial wall remodeling. 5-LO is expressed by leukocytes including blood monocytes, tissue macrophages, dendritic cells, neutrophils, and mast cells. LTB4 and the cysteinyl LTs LTC4, LTD4, and LTE4, act through two BLT and two cysLT receptors that are differentially expressed on hematopoietic and arterial wall cells. The precise roles of LTs or the LT receptors in cardiovascular physiology remain largely to be explored. In this review, we will discuss what is currently known about the 5-LO atherosclerosis connection. We will attempt to propose strategies to further explore potential links between the 5-LO pathway and blood vessel physiology and disease progression.


Asunto(s)
Araquidonato 5-Lipooxigenasa/fisiología , Arterias/enzimología , Arteriosclerosis/enzimología , Transducción de Señal/fisiología , Animales , Arteriosclerosis/genética , Ratones , Transducción de Señal/genética
6.
Arterioscler Thromb Vasc Biol ; 23(8): e32-6, 2003 Aug 01.
Artículo en Inglés | MEDLINE | ID: mdl-12816882

RESUMEN

OBJECTIVE: Inflammatory infiltrates and atherosclerotic lesions emerge when monocytes adhere to endothelial cells (ECs), migrate into the subendothelial space, and become macrophages (MPhi(s)). Leukotrienes (LTs), products of 5-lipoxygenase, are powerful inflammatory mediators. 5-lipoxygenase+ MPhi(s) have been shown to increase during atherogenesis, and LT receptor (LT-R) transcripts were identified in diseased arteries. To investigate LT-Rs in cells involved in inflammation and atherogenesis, we used the in vitro models of human umbilical vein ECs (HUVECs) and monocyte-derived MPhi(s). METHODS AND RESULTS: HUVECs primarily expressed transcripts of the cysteinyl (cys) LT2-R, which was strongly upregulated by interleukin-4. By contrast, MPhi(s) predominantly expressed transcripts of the cysLT1-R. Calcium responses toward LTs revealed differential cysLT-R utilization by both cell types: HUVECs responded to both cysLTs, whereas MPhi(s) preferentially responded to LTD4; HUVECs, but not MPhi(s), were resistant toward a cysLT1-R antagonist, montelukast; cysLTs generated regular calcium oscillations in HUVECs that lasted >60 minutes, resulting in >500 oscillations per cell. By contrast, calcium elevations in MPhi(s) returned to baseline within seconds and were nonoscillatory. CONCLUSIONS: Our data raise the possibility that MPhi-derived LTs differentially activate cysLT2-Rs via paracrine stimulation and cysLT1-Rs via autocrine and paracrine stimulation during inflammation and atherogenesis.


Asunto(s)
Araquidonato 5-Lipooxigenasa/metabolismo , Arteriosclerosis/metabolismo , Calcio/metabolismo , Endotelio Vascular/metabolismo , Macrófagos/metabolismo , Receptores de Leucotrienos/genética , Receptores de Leucotrienos/metabolismo , Arteriosclerosis/etiología , Células Cultivadas , Perfilación de la Expresión Génica , Humanos , Inflamación/complicaciones , Inflamación/fisiopatología , Regulación hacia Arriba
7.
Thromb Haemost ; 90(4): 704-9, 2003 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-14515192

RESUMEN

The present study investigates whether vascular smooth muscle cells of the human saphenous vein (SMC) express a functionally active protease-activated receptor-3 (PAR-3). PAR-3 mRNA was detected by RT-PCR. In the presence of thrombin, a rapid and transient increase in PAR-3 mRNA was observed. Stimulation of SMC with thrombin or the synthetic PAR-3-activating peptide, TFRGAP, resulted in transient mobilization of intracellular calcium. After a preceding challenge with thrombin, the calcium signal to TFRGAP was abolished, suggesting cleavage and subsequent desensitization of PAR-3 by thrombin. Activation of PAR-3 by TFRGAP elicited a time-dependent activation of the extracellular-signal-regulated kinase (ERK)-1/2 with a maximum response 10-20 min after stimulation. At 200 microM, TFRGAP increased [3H]-thymidine incorporation into cellular DNA about two-fold. These data indicate that PAR-3 is expressed in human SMC and triggers intracellular signaling. Thus, in the SMC PAR-3 might contribute to thrombin-induced responses.


Asunto(s)
Músculo Liso Vascular/citología , Miocitos del Músculo Liso/química , Receptores de Trombina/análisis , Señalización del Calcio , Replicación del ADN/efectos de los fármacos , Humanos , Cinética , Proteína Quinasa 3 Activada por Mitógenos , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Músculo Liso Vascular/química , ARN Mensajero/análisis , ARN Mensajero/efectos de los fármacos , Receptores de Trombina/genética , Receptores de Trombina/fisiología , Vena Safena , Trombina/farmacología
8.
J Exp Med ; 206(1): 233-48, 2009 Jan 16.
Artículo en Inglés | MEDLINE | ID: mdl-19139167

RESUMEN

Atherosclerosis involves a macrophage-rich inflammation in the aortic intima. It is increasingly recognized that this intimal inflammation is paralleled over time by a distinct inflammatory reaction in adjacent adventitia. Though cross talk between the coordinated inflammatory foci in the intima and the adventitia seems implicit, the mechanism(s) underlying their communication is unclear. Here, using detailed imaging analysis, microarray analyses, laser-capture microdissection, adoptive lymphocyte transfers, and functional blocking studies, we undertook to identify this mechanism. We show that in aged apoE(-/-) mice, medial smooth muscle cells (SMCs) beneath intimal plaques in abdominal aortae become activated through lymphotoxin beta receptor (LTbetaR) to express the lymphorganogenic chemokines CXCL13 and CCL21. These signals in turn trigger the development of elaborate bona fide adventitial aortic tertiary lymphoid organs (ATLOs) containing functional conduit meshworks, germinal centers within B cell follicles, clusters of plasma cells, high endothelial venules (HEVs) in T cell areas, and a high proportion of T regulatory cells. Treatment of apoE(-/-) mice with LTbetaR-Ig to interrupt LTbetaR signaling in SMCs strongly reduced HEV abundance, CXCL13, and CCL21 expression, and disrupted the structure and maintenance of ATLOs. Thus, the LTbetaR pathway has a major role in shaping the immunological characteristics and overall integrity of the arterial wall.


Asunto(s)
Aorta Abdominal/crecimiento & desarrollo , Apolipoproteínas E/genética , Tejido Conectivo/crecimiento & desarrollo , Receptor beta de Linfotoxina/fisiología , Transducción de Señal/fisiología , Envejecimiento , Animales , Aorta Abdominal/metabolismo , Aterosclerosis/genética , Transporte Biológico , Células Cultivadas , Quimiocina CCL21/genética , Quimiocina CCL21/metabolismo , Quimiocina CXCL13/genética , Quimiocina CXCL13/metabolismo , Análisis por Conglomerados , Tejido Conectivo/metabolismo , Perfilación de la Expresión Génica , Hibridación in Situ , Leucocitos Mononucleares/citología , Leucocitos Mononucleares/metabolismo , Tejido Linfoide/citología , Tejido Linfoide/crecimiento & desarrollo , Tejido Linfoide/metabolismo , Receptor beta de Linfotoxina/antagonistas & inhibidores , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Miocitos del Músculo Liso/citología , Miocitos del Músculo Liso/metabolismo , Organogénesis , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Túnica Íntima/crecimiento & desarrollo , Túnica Íntima/metabolismo , Túnica Media/crecimiento & desarrollo , Túnica Media/metabolismo
9.
Proc Natl Acad Sci U S A ; 103(16): 6326-31, 2006 Apr 18.
Artículo en Inglés | MEDLINE | ID: mdl-16606835

RESUMEN

Cysteinyl leukotrienes (cysLT), i.e., LTC4, LTD4, and LTE4, are lipid mediators derived from the 5-lipoxygenase pathway, and the cysLT receptors cysLT1-R/cysLT2-R mediate inflammatory tissue reactions. Although endothelial cells (ECs) predominantly express cysLT2-Rs, their role in vascular biology remains to be fully understood. To delineate cysLT2-R actions, we stimulated human umbilical vein EC with LTD4 and determined early induced genes. We also compared LTD4 effects with those induced by thrombin that binds to protease-activated receptor (PAR)-1. Stringent filters yielded 37 cysLT2-R- and 34 PAR-1-up-regulated genes (>2.5-fold stimulation). Most LTD4-regulated genes were also induced by thrombin. Moreover, LTD4 plus thrombin augmented gene expression when compared with each agonist alone. Strongly induced genes were studied in detail: Early growth response (EGR) and nuclear receptor subfamily 4 group A transcription factors; E-selectin; CXC ligand 2; IL-8; a disintegrin-like and metalloprotease (reprolysin type) with thrombospondin type 1 motif 1 (ADAMTS1); Down syndrome critical region gene 1 (DSCR1); tissue factor (TF); and cyclooxygenase 2. Transcripts peaked at approximately 60 min, were unaffected by a cysLT1-R antagonist, and were superinduced by cycloheximide. The EC phenotype was markedly altered: LTD4 induced de novo synthesis of EGR1 protein and EGR1 localized in the nucleus; LTD4 up-regulated IL-8 formation and secretion; and LTD4 raised TF protein and TF-dependent EC procoagulant activity. These data show that cysLT2-R activation results in a proinflammatory EC phenotype. Because LTD4 and thrombin are likely to be formed concomitantly in vivo, cysLT2-R and PAR-1 may cooperate to augment vascular injury.


Asunto(s)
Endotelio Vascular/metabolismo , Regulación de la Expresión Génica , Leucotrieno C4/metabolismo , Proteínas de la Membrana/fisiología , Receptor PAR-1/fisiología , Receptores de Leucotrienos/fisiología , Trombina/metabolismo , Coagulación Sanguínea/efectos de los fármacos , Coagulación Sanguínea/fisiología , Células Cultivadas , Células Endoteliales/efectos de los fármacos , Células Endoteliales/metabolismo , Endotelio Vascular/efectos de los fármacos , Humanos , Leucotrieno C4/farmacología , Proteínas de la Membrana/agonistas , Receptor PAR-1/agonistas , Receptores Citoplasmáticos y Nucleares , Receptores de Leucotrienos/agonistas , Trombina/farmacología , Transcripción Genética/efectos de los fármacos , Venas Umbilicales/citología
10.
Proc Natl Acad Sci U S A ; 100(3): 1238-43, 2003 Feb 04.
Artículo en Inglés | MEDLINE | ID: mdl-12552108

RESUMEN

Oxidation products of low-density lipoproteins have been suggested to promote inflammation during atherogenesis, and reticulocyte-type 15-lipoxygenase has been implicated to mediate this oxidation. In addition, the 5-lipoxygenase cascade leads to formation of leukotrienes, which exhibit strong proinflammatory activities in cardiovascular tissues. Here, we studied both lipoxygenase pathways in human atherosclerosis. The 5-lipoxygenase pathway was abundantly expressed in arterial walls of patients afflicted with various lesion stages of atherosclerosis of the aorta and of coronary and carotid arteries. 5-lipoxygenase localized to macrophages, dendritic cells, foam cells, mast cells, and neutrophilic granulocytes, and the number of 5-lipoxygenase expressing cells markedly increased in advanced lesions. By contrast, reticulocyte-type 15-lipoxygenase was expressed at levels that were several orders of magnitude lower than 5-lipoxygenase in both normal and diseased arteries, and its expression could not be related to lesion pathology. Our data support a model of atherogenesis in which 5-lipoxygenase cascade-dependent inflammatory circuits consisting of several leukocyte lineages and arterial wall cells evolve within the blood vessel wall during critical stages of lesion development. They raise the possibility that antileukotriene drugs may be an effective treatment regimen in late-stage disease.


Asunto(s)
Araquidonato 5-Lipooxigenasa/metabolismo , Arterias/enzimología , Antígenos CD/biosíntesis , Antígenos de Diferenciación Mielomonocítica/biosíntesis , Aorta/enzimología , Araquidonato 15-Lipooxigenasa/metabolismo , Araquidonato 5-Lipooxigenasa/biosíntesis , Arteriosclerosis/patología , Linaje de la Célula , Endotelio Vascular/enzimología , Humanos , Immunoblotting , Inmunohistoquímica , Cinética , Leucocitos/enzimología , Macrófagos/enzimología , Fenotipo , ARN/metabolismo , ARN Mensajero/metabolismo , Reticulocitos/enzimología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Distribución Tisular
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA