RESUMEN
Although neutrophils play critical roles in innate immunity, in excess these cells cause severe tissue damage. Thus, neutrophil activation must be tightly regulated to prevent indiscriminant damage. Previously, we reported that mice lacking matrix metalloproteinase (MMP) 7 are protected from lung injury owing to markedly impaired neutrophil movement from the interstitium into mucosal lumenal spaces. This phenotype resulted from a lack of MMP7 shedding of syndecan-1, a heparan sulfate proteoglycan that carries the neutrophil chemokine CXCL1 as cargo. Here, we assessed if shedding syndecan-1/CXCL1 complexes affects neutrophil activation. Whereas injured monolayers of wild-type alveolar type II cells potently stimulated neutrophil activation, as gauged by release of myeloperoxidase, cells from Mmp7(-/-) or syndecan-1-null (Sdc1(-/-)) mice or human cells with MMP7 knockdown did not. In vivo, we observed reduced myeloperoxidase release relative to neutrophil numbers in bleomycin-injured Mmp7(-/-) and Sdc1(-/-) mice. Furthermore, we determined that soluble syndecan-1 directly stimulated neutrophil activation in the absence of cellular damage. These data indicate that MMP7 shedding of syndecan-1/CXCL1 complexes functions as a checkpoint that restricts neutrophil activation at sites of epithelial injury.
Asunto(s)
Quimiocina CXCL1/metabolismo , Células Epiteliales/metabolismo , Metaloproteinasa 7 de la Matriz/metabolismo , Activación Neutrófila , Sindecano-1/metabolismo , Células Epiteliales Alveolares/metabolismo , Células Epiteliales Alveolares/patología , Animales , Femenino , Masculino , Ratones Endogámicos C57BL , Modelos Biológicos , Neutrófilos/metabolismoRESUMEN
Obesity is a strong risk factor for the development of type 2 diabetes. We have previously reported that in adipose tissue of obese (ob/ob) mice, the expression of adipogenic genes is decreased. When made genetically obese, the BTBR mouse strain is diabetes susceptible and the C57BL/6J (B6) strain is diabetes resistant. We used DNA microarrays and RT-PCR to compare the gene expression in BTBR-ob/ob versus B6-ob/ob mice in adipose tissue, liver, skeletal muscle, and pancreatic islets. Our results show: 1) there is an increased expression of genes involved in inflammation in adipose tissue of diabetic mice; 2) lipogenic gene expression was lower in adipose tissue of diabetes-susceptible mice, and it continued to decrease with the development of diabetes, compared with diabetes-resistant obese mice; 3) hepatic expression of lipogenic enzymes was increased and the hepatic triglyceride content was greatly elevated in diabetes-resistant obese mice; 4) hepatic expression of gluconeogenic genes was suppressed at the prediabetic stage but not at the onset of diabetes; and 5) genes normally not expressed in skeletal muscle and pancreatic islets were expressed in these tissues in the diabetic mice. We propose that increased hepatic lipogenic capacity protects the B6-ob/ob mice from the development of type 2 diabetes.
Asunto(s)
Diabetes Mellitus/genética , Perfilación de la Expresión Génica , Predisposición Genética a la Enfermedad , Lípidos/biosíntesis , Hígado/metabolismo , Obesidad/genética , Tejido Adiposo/metabolismo , Animales , Diabetes Mellitus Tipo 2/genética , Hígado Graso/genética , Femenino , Gluconeogénesis/genética , Islotes Pancreáticos/metabolismo , Hígado/enzimología , Ratones , Ratones Endogámicos C57BL , Ratones Obesos , Músculo Esquelético/metabolismo , Análisis de Secuencia por Matrices de Oligonucleótidos , ARN Mensajero/análisis , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Triglicéridos/sangre , Triglicéridos/metabolismoRESUMEN
The advent of sophisticated genomic techniques for gene mapping and microarray analysis has provided opportunities to map mRNA abundance to quantitative trait loci (QTL) throughout the genome. Unfortunately, simple mapping of each individual mRNA trait on the scale of a typical microarray experiment is computationally intensive, subject to high sample variance, and therefore underpowered. However, this problem can be addressed by capitalizing on correlation among the large number of mRNA traits. We present a method to reduce the dimensionality for mapping gene expression data as quantitative traits. We used a blind method, principal components, and a sighted method, hierarchical clustering seeded by disease relevant traits, to define new traits composed of a small collection of promising mRNAs. We validated the principle of our approach by mapping the expression levels of metabolism genes in a population of F(2)-ob/ob mice derived from the BTBR and C57BL/6J strains. We found that lipogenic and gluconeogenic mRNAs, which are known targets of insulin action, were closely associated with the insulin trait. Multiple interval mapping and Bayesian interval mapping of this new trait revealed significant linkages to chromosome regions that were contained in loci associated with type 2 diabetes in this same mouse sample. As a further statistical refinement, we show that principal component analysis also effectively reduced dimensions for mapping phenotypes composed of mRNA abundances.
Asunto(s)
Glucemia/genética , Mapeo Cromosómico , Carácter Cuantitativo Heredable , ARN Mensajero/análisis , Animales , Teorema de Bayes , Glucemia/análisis , Índice de Masa Corporal , Análisis por Conglomerados , Ayuno , Femenino , Expresión Génica , Ligamiento Genético , Insulina/genética , Funciones de Verosimilitud , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos , Ratones Obesos , Repeticiones de Microsatélite , Reacción en Cadena de la Polimerasa , Sitios de Carácter Cuantitativo , Reproducibilidad de los ResultadosRESUMEN
Multiple conditions result in development of pulmonary hypertension. Pulmonary arterial hypertension (PAH) is the subclassification of pulmonary hypertension, in which known or unknown underlying conditions lead to similar intrinsic alterations in the pulmonary vasculature. PAH is a progressive condition characterized by restricted blood flow through the pulmonary circulation leading to poor survival in the absence of effective therapy. Over the last two decades, new therapeutic agents have substantially improved the course and prognosis for PAH patients. Three available classes of drugs, ie, prostacyclins, endothelin receptor antagonists, and phosphodiesterase-5 inhibitors provide multiple options for treatment of PAH. Endothelin receptor antagonists and phosphodiesterase-5 inhibitors are administered orally, whereas prostacyclin therapies are delivered by continuous intravenous or subcutaneous infusion, or as aerosols by nebulization. Because of the risks and inconveniences associated with administration, prostacyclins are typically reserved for patients with more advanced disease or progression despite oral therapy. Inhaled administration may be a safer and easier route for prostacyclin administration. Treprostinil is a prostacyclin analog that has been demonstrated to be effective when administered by continuous subcutaneous or intravenous infusion, and more recently by nebulization.
Asunto(s)
Antihipertensivos/administración & dosificación , Epoprostenol/análogos & derivados , Hipertensión Pulmonar/tratamiento farmacológico , Administración por Inhalación , Administración Oral , Epoprostenol/administración & dosificación , Epoprostenol/farmacología , Hipertensión Pulmonar Primaria Familiar , Humanos , Hipertensión Pulmonar/diagnóstico , Hipertensión Pulmonar/epidemiologíaRESUMEN
BACKGROUND: Lung injury promotes the expression of matrix metalloproteinase-7 (MMP7, matrilysin), which is required for neutrophil recruitment and re-epithelialization. MMP7 governs the lung inflammatory response through the shedding of syndecan-1. Because inflammation and repair are related events, we evaluated the role of syndecan-1 shedding in lung re-epithelialization. METHODOLOGY/PRINCIPAL FINDING: Epithelial injury induced syndecan-1 shedding from wild-type epithelium but not from Mmp7(-/-) mice in vitro and in vivo. Moreover, cell migration and wound closure was enhanced by MMP7 shedding of syndecan-1. Additionally, we found that syndecan-1 augmented cell adhesion to collagen by controlling the affinity state of the alpha(2)beta(1) integrin. CONCLUSION/SIGNIFICANCE: MMP7 shedding of syndecan-1 facilitates wound closure by causing the alpha(2)beta(1) integrin to assume a less active conformation thereby removing restrictions to migration. MMP7 acts in the lungs to regulate inflammation and repair, and our data now show that both these functions are controlled through the shedding of syndecan-1.
Asunto(s)
División Celular/fisiología , Integrina alfa2beta1/metabolismo , Pulmón/enzimología , Metaloproteinasa 7 de la Matriz/metabolismo , Sindecano-1/metabolismo , Animales , Adhesión Celular/fisiología , Movimiento Celular , Células Epiteliales/citología , Técnicas de Silenciamiento del Gen , Pulmón/citología , Ratones , Ratones Endogámicos C57BL , Sindecano-1/fisiologíaRESUMEN
Insulin resistance is a common feature of obesity. BTBR mice have more fat mass than most other inbred mouse strains. On a chow diet, BTBR mice have elevated insulin levels relative to the C57BL/6J (B6) strain. Male F1 progeny of a B6 x BTBR cross are insulin resistant. Previously, we reported insulin resistance in isolated muscle and in isolated adipocytes in this strain. Whereas the muscle insulin resistance was observed only in male F1 mice, adipocyte insulin resistance was also present in male BTBR mice. We examined in vivo mechanisms of insulin resistance with the hyperinsulinemic euglycemic clamp technique. At 10 wk of age, BTBR and F1 mice had a >30% reduction in whole body glucose disposal primarily due to insulin resistance in heart, soleus muscle, and adipose tissue. The increased adipose tissue mass and decreased muscle mass in BTBR and F1 mice were negatively and positively correlated with whole body glucose disposal, respectively. Genes involved in focal adhesion, actin cytoskeleton, and inflammation were more highly expressed in BTBR and F1 than in B6 adipose tissue. The BTBR and F1 mice have higher levels of testosterone, which may be related to the pathological changes in adipose tissue that lead to systemic insulin resistance. Despite profound peripheral insulin resistance, BTBR and F1 mice retained hepatic insulin sensitivity. These studies reveal a genetic difference in body composition that correlates with large differences in peripheral insulin sensitivity.
Asunto(s)
Grasa Abdominal , Resistencia a la Insulina , Insulina/sangre , Hígado/metabolismo , Obesidad/metabolismo , Grasa Abdominal/metabolismo , Animales , Composición Corporal , Femenino , Expresión Génica , Glucosa/metabolismo , Técnica de Clampeo de la Glucosa , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos , Obesidad/sangreRESUMEN
The BTBR mouse strain harbors alleles promoting insulin resistance. When made genetically obese (ob/ob), these mice develop severe type 2 diabetes (fasting glucose >400 mg/dL). By contrast, C57BL/6 ob/ob mice are able to compensate for the obesity-induced insulin resistance by increasing pancreatic insulin secretion and thus maintain only slightly elevated plasma glucose levels (<250 mg/dL). Islet insulin secretory responses to glucose are undiminished in the remaining islets of BTBR ob/ob mice. A genome-wide linkage analysis identified 3 major loci influencing plasma glucose and/or insulin levels in an F2ob/ob sample derived from the 2 strains. A locus on chromosome 2 affects insulin sensitivity and is independent of obesity. Loci on chromosomes 16 and 19 affect fasting glucose and insulin levels and likely affect beta-cell mass or function. Analysis of mRNA expression patterns revealed a reduction in lipogenic gene expression in adipose tissue associated with obesity. Conversely, hepatic lipogenic gene expression increases in obese mice, but to a much greater extent in the diabetes-resistant C57BL/6 strain. We propose that hepatic lipogenic capacity affects susceptibility to obesity-induced diabetes.