RESUMEN
OBJECTIVE: ATP-binding cassette transporter subfamily G member 2 (ABCG2), expressed in microvascular endothelial cells in the heart, has been suggested to regulate several tissue defense mechanisms. This study was performed to elucidate its role in pressure overload-induced cardiac hypertrophy. METHODS AND RESULTS: Pressure overload was induced in 8- to 12-week-old wild-type and Abcg2-/- mice by transverse aortic constriction (TAC). Abcg2-/- mice showed exaggerated cardiac hypertrophy and ventricular remodeling after TAC compared with wild-type mice. In the early phase after TAC, functional impairment in angiogenesis and antioxidant response in myocardium was found in Abcg2-/- mice. In vitro experiments demonstrated that ABCG2 regulates transport of glutathione, an important endogenous antioxidant, from microvascular endothelial cells. Besides, glutathione transported from microvascular endothelial cells in ABCG2-dependent manner ameliorated oxidative stress-induced cardiomyocyte hypertrophy. In vivo, glutathione levels in plasma and the heart were increased in wild-type mice but not in Abcg2-/- mice after TAC. Treatment with the superoxide dismutase mimetic ameliorated cardiac hypertrophy in Abcg2-/- mice after TAC to the same extent as that in wild-type mice, although cardiac dysfunction with impaired angiogenesis was observed in Abcg2-/- mice. CONCLUSION: ABCG2 protects against pressure overload-induced cardiac hypertrophy and heart failure by promoting angiogenesis and antioxidant response.
Asunto(s)
Transportadoras de Casetes de Unión a ATP/metabolismo , Antioxidantes/metabolismo , Células Endoteliales/metabolismo , Glutatión/metabolismo , Insuficiencia Cardíaca/prevención & control , Hipertrofia Ventricular Izquierda/prevención & control , Miocitos Cardíacos/metabolismo , Proteínas de Neoplasias/metabolismo , Neovascularización Fisiológica , Estrés Oxidativo , Transportador de Casetes de Unión a ATP, Subfamilia G, Miembro 2 , Transportadoras de Casetes de Unión a ATP/genética , Animales , Animales Recién Nacidos , Antioxidantes/farmacología , Células Cultivadas , Modelos Animales de Enfermedad , Células Endoteliales/efectos de los fármacos , Genotipo , Glutatión/sangre , Insuficiencia Cardíaca/genética , Insuficiencia Cardíaca/metabolismo , Insuficiencia Cardíaca/fisiopatología , Miembro Posterior , Humanos , Hipertrofia Ventricular Izquierda/genética , Hipertrofia Ventricular Izquierda/metabolismo , Hipertrofia Ventricular Izquierda/fisiopatología , Isquemia/genética , Isquemia/metabolismo , Isquemia/fisiopatología , Masculino , Ratones , Ratones Noqueados , Músculo Esquelético/irrigación sanguínea , Miocitos Cardíacos/efectos de los fármacos , Proteínas de Neoplasias/genética , Estrés Oxidativo/efectos de los fármacos , Fenotipo , Interferencia de ARN , Ratas , Ratas Wistar , Factores de Tiempo , Transfección , Función Ventricular , Remodelación VentricularRESUMEN
OBJECTIVE: To clarify the impact of breast cancer resistance protein 1 (BCRP1)/ATP-binding cassette transporter subfamily G member 2 (ABCG2) expression on cardiac repair after myocardial infarction (MI). METHODS AND RESULTS: The ATP-binding cassette transporter BCRP1/ABCG2 is expressed in various organs, including the heart, and may regulate several tissue defense mechanisms. BCRP1/ABCG2 was mainly expressed in endothelial cells of microvessels in the heart. MI was induced in 8- to 12-week-old wild-type (WT) and Bcrp1/Abcg2 knockout (KO) mice by ligating the left anterior descending artery. At 28 days after MI, the survival rate was significantly lower in KO mice than in WT mice because of cardiac rupture. Echocardiographic, hemodynamic, and histological assessments showed that ventricular remodeling was more deteriorated in KO than in WT mice. Capillary, myofibroblast, and macrophage densities in the peri-infarction area at 5 days after MI were significantly reduced in KO compared with WT mice. In vitro experiments demonstrated that inhibition of BCRP1/ABCG2 resulted in accumulation of intracellular protoporphyrin IX and impaired survival of microvascular endothelial cells under oxidative stress. Moreover, BCRP1/ABCG2 inhibition impaired migration and tube formation of endothelial cells. CONCLUSIONS: BCRP1/ABCG2 plays a pivotal role in cardiac repair after MI via modulation of microvascular endothelial cell survival and function.
Asunto(s)
Transportadoras de Casetes de Unión a ATP/fisiología , Células Endoteliales/fisiología , Microvasos/fisiopatología , Infarto del Miocardio/fisiopatología , Proteínas de Neoplasias/fisiología , Transportador de Casetes de Unión a ATP, Subfamilia G, Miembro 2 , Animales , Supervivencia Celular , Femenino , Ratones , Ratones Noqueados , Recuperación de la Función , Cicatrización de HeridasRESUMEN
OBJECTIVE: Circulating progenitors and stem cells have been reported to contribute to angiogenesis and arterial repair after injury. In the present study, we investigated whether the arterial wall could host permanently residing progenitor cells under physiological context. METHODS AND RESULTS: Using the Hoechst-based flow cytometry method, we identified and isolated progenitor cells termed side population (SP) cells at a prevalence of 6.0+/-0.8% in the tunica media of adult mice aortas. Arterial SP cells expressed the ATP-binding cassette transporter subfamily G member 2, frequently present on SP cell surface, and displayed a Sca-1+ c-kit(-/low) Lin- CD34(-/low) profile. They did not form myeloid or lymphoid hematopoietic colonies after plating in methylcellulose-based medium. Importantly, cultured SP cells were able to acquire the phenotype of endothelial cells (CD31, VE-cadherin, and von Willebrand factor expression) or of smooth muscle cells (alpha-smooth muscle actin, calponin, and smooth muscle myosin heavy chain expression), in presence of either vascular endothelial growth factor or transforming growth factor (TGF)-beta1/PDGF-BB, respectively. Moreover, they generated vascular-like branching structures, composed of both VE-cadherin+ cells and alpha-smooth muscle actin+ cells on Matrigel. CONCLUSIONS: In this study, we provide the first evidence to our knowledge that in the adult mice, the normal arterial wall harbors SP cells with vascular progenitor properties.
Asunto(s)
Aorta Abdominal/citología , Aorta Torácica/citología , Separación Celular/métodos , Células Madre/citología , Transportador de Casetes de Unión a ATP, Subfamilia G, Miembro 2 , Transportadoras de Casetes de Unión a ATP/metabolismo , Factores de Edad , Animales , Materiales Biocompatibles , Biomarcadores , Técnicas de Cultivo de Célula/métodos , Diferenciación Celular , Colágeno , Combinación de Medicamentos , Femenino , Citometría de Flujo , Laminina , Metilcelulosa , Ratones , Ratones Endogámicos C57BL , Fenotipo , Proteoglicanos , Células Madre/metabolismo , Túnica Media/citologíaRESUMEN
Atherosclerosis results from accumulation of macrophages and extracellular matrix in the arterial wall. Decorin, a small matrix proteoglycan, is able to regulate cell proliferation, migration and growth factors' activity. We investigated the effect of decorin overexpression on atherosclerosis progression in apolipoprotein E-deficient (ApoE(-/-)) mice. Female ApoE(-/-) mice, 10 weeks old (early treatment, n = 20) and 20 weeks old (delayed treatment, n = 20) were administered intravenously with either an adenovirus (2.5 x 10(9) plaque-forming units/mouse) containing human decorin gene (Ad-Dcn) or beta-galactosidase (LacZ), or PBS. Transgenic decorin was mainly expressed in the liver, and was secreted in the plasma up to 4 weeks. Six weeks after treatment, no significant difference in aortic root lesion size was observed between LacZ- and PBS-control groups. In contrast, Ad-Dcn-treated mice showed significantly reduced atherosclerotic lesions as compared to controls in both early and delayed treatment groups (2.9 +/- 1.1% versus 5.5 +/- 0.4%; p = 0.004 and 13.4 +/- 1.3% versus 19.9 +/- 1.41%; p = 0.009, respectively). In parallel, macrophage, gelatinase activity and collagen plaque content were also reduced. Interestingly, plasma triglycerides were reduced and decorin formed complexes with transforming growth factor-beta1 (TGF-beta1) that resulted in reduced circulating free-TGF-beta1. In conclusion, systemic overexpression of decorin reduces inflammation, triglycerides and fibrosis in atherosclerotic plaques of ApoE(-/-) mice resulting in slowing down of disease progression.
Asunto(s)
Apolipoproteínas E/genética , Aterosclerosis/metabolismo , Proteínas de la Matriz Extracelular/biosíntesis , Proteoglicanos/biosíntesis , Amiloide/sangre , Animales , Apolipoproteínas E/fisiología , Proliferación Celular , Decorina , Progresión de la Enfermedad , Matriz Extracelular/metabolismo , Proteínas de la Matriz Extracelular/metabolismo , Terapia Genética/métodos , Inflamación , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Proteoglicanos/metabolismo , ARN/metabolismo , Proteínas Recombinantes/metabolismo , Factores de Tiempo , Triglicéridos/metabolismoAsunto(s)
Endotelio Vascular/metabolismo , Epoprostenol/metabolismo , Células Madre Mesenquimatosas/metabolismo , Neovascularización Fisiológica/fisiología , Túnica Íntima/metabolismo , Animales , Endotelio Vascular/citología , Células Madre Mesenquimatosas/citología , Ratones , Ratones Noqueados , Modelos Animales , Receptores de Epoprostenol/genética , Receptores de Epoprostenol/metabolismo , Transducción de Señal/fisiología , Túnica Íntima/citologíaRESUMEN
OBJECTIVE: Lymphocyte activation is thought to play a major role in the pathogenesis of atherosclerotic complications such as plaque thrombosis. Circulating CD31+ T cells have been shown to regulate human T cell activation. Aim of this study was to evaluate whether the proportion of circulating immunoregulatory CD31+ T cells is correlated to the occurrence of plaque thrombosis in aged apolipoprotein (apo) E knockout (KO) mice. METHODS AND RESULTS: CD31+ T cell depletion of spleen T cells enhanced proliferation (P<0.05) and interferon-gamma production (P<0.01) while reducing interleukin (IL)-4 (P<0.001) and IL-10 (P=0.001) secretion in response to minimally modified low-density lipoprotein. CD31+ T cells were counted in 65 apoE KO mice (46-week-old) by flow cytometry. Organizing thrombi could be documented in 28 of 195 (14%) lesions and in at least one of the aorta root lesions in 23 of 65 mice (35%). CD31+ T cell count was significantly reduced in mice showing plaque thrombosis (72.3+/-1.5% versus 84.1+/-1.2%; P<0.0001), but such reduction did not follow induced plaque rupture or experimentally controlled thrombosis. CONCLUSIONS: Reduced CD31+ T cells in circulating blood is a hallmark of atherosclerotic plaque thrombosis. Our data suggest that CD31+ T cells may play an important regulatory role in the development of plaque thrombosis.
Asunto(s)
Aterosclerosis/inmunología , Activación de Linfocitos/inmunología , Linfocitos T/inmunología , Trombosis/inmunología , Animales , Apolipoproteínas E/genética , Aterosclerosis/patología , Biomarcadores , Femenino , Inmunohistoquímica , Recuento de Linfocitos , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Molécula-1 de Adhesión Celular Endotelial de Plaqueta/metabolismo , Rotura , Linfocitos T/citología , Linfocitos T/metabolismo , Trombosis/patologíaRESUMEN
Administration of monoclonal antibodies directed against the leukocyte function-associated antigen 1 (LFA-1)-intercellular adhesion molecule 1 (ICAM-1) pathway showed that these costimulatory molecules play a key role in allograft rejection. Here, adenoviral gene transfer of an immunoadhesin, sICAM-1/Ig, was used to prolong islet allograft survival in a mouse model, and was compared with anti-LFA-1 antibody administration. A replication-deficient recombinant adenoviral vector encoding a chimeric protein, in which the extracellular domain of ICAM-1 is covalently linked to the C(H)2-C(H)3 domains of an IgG1, was used for gene transfer. C3H murine islets were transplanted under the kidney capsule of streptozotocin-induced diabetic BALB/c mice. Experimental groups underwent adenovirus vector administration either in vivo (intravenous injection) or ex vivo (gene transfer to the graft), and control groups received either an empty vector (Ad.null) or an anti-LFA-1 monoclonal antibody. Graft survival was significantly prolonged by in vivo sICAM-1/Ig gene transfer as compared with both Ad.null and anti-LFA-1 groups, but not by ex vivo gene transfer. Histological examination of the grafts showed the presence of a mononuclear infiltrate within functioning grafts, suggesting that the homing of alloreactive T cells was not altered. In vitro T cell proliferation experiments indicated that sICAM-1/Ig exerted agonist effects on both CD4(+) and CD8(+) T cells.
Asunto(s)
Adenoviridae , Técnicas de Transferencia de Gen , Vectores Genéticos , Supervivencia de Injerto/genética , Inmunoglobulina G/genética , Molécula 1 de Adhesión Intercelular/genética , Trasplante de Islotes Pancreáticos/inmunología , Animales , Anticuerpos Monoclonales/administración & dosificación , Molécula 1 de Adhesión Intercelular/inmunología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C3H , Proteínas Recombinantes de Fusión , Linfocitos T/inmunologíaRESUMEN
OBJECTIVE: One of the early hallmarks of osteoarthritis (OA) is a progressive degeneration of the articular cartilage. Early diagnosis of OA-associated cartilage alterations would be beneficial for disease prevention and control, and for the development of disease-modifying treatments. However, early diagnosis is still hampered by a lack of quantifiable readouts in preclinical models. DESIGN: In this study, we have shown the potency of contrast-enhanced nanofocus x-ray computed tomography (CE-nanoCT) to be used for virtual 3-dimensional (3D) histopathology in established mouse models for OA, and we compared with standard histopathology. RESULTS: We showed the equivalence of CE-nanoCT images to histopathology for the modified Mankin scoring of the cartilage structure and quality. Additionally, a limited set of 3D cartilage characteristics measured by CE-nanoCT image analysis in a user-independent and semiautomatic manner, that is, average and maximum of the noncalcified cartilage thickness distribution and loss in glycosaminoglycans, was shown to be predictive for the cartilage quality and structure as can be evaluated by histopathological scoring through the use of an empirical model. CONCLUSIONS: We have shown that CE-nanoCT is a tool that allows virtual histopathology and 3D morphological quantification of multitissue systems, such as the chondro-osseous junction. It provides faster and more quantitative data on cartilage structure and quality compared with standard histopathology while eliminating user bias. CE-nanoCT thus should allow capturing subtle differences in cartilage characteristics, carefully mapping OA progression and, ultimately, asses the beneficial changes when testing a candidate disease-modifying treatment.