RESUMO
Abdominal aortic aneurysm (AAA) pathogenesis is distinguished by vessel wall inflammation. Cyclooxygenase (COX)-2 and microsomal prostaglandin E synthase-1, key components of the most well-characterized inflammatory prostaglandin pathway, contribute to AAA development in the 28-day angiotensin II infusion model in mice. In this study, we used this model to examine the role of the prostaglandin E receptor subtype 4 (EP4) and genetic knockdown of COX-2 expression (70% to 90%) in AAA pathogenesis. The administration of the prostaglandin receptor EP4 antagonist AE3-208 (10 mg/kg per day) to apolipoprotein E (apoE)-deficient mice led to active drug plasma concentrations and reduced AAA incidence and severity compared with control apoE-deficient mice (P < 0.01), whereas COX-2 genetic knockdown/apoE-deficient mice displayed only a minor, nonsignificant decrease in incidence of AAA. EP4 receptor protein was present in human and mouse AAA, as observed by using Western blot analysis. Aortas from AE3-208-treated mice displayed evidence of a reduced inflammatory phenotype compared with controls. Atherosclerotic lesion size at the aortic root was similar between all groups. In conclusion, the prostaglandin E(2)-EP4 signaling pathway plays a role in the AAA inflammatory process. Blocking the EP4 receptor pharmacologically reduces both the incidence and severity of AAA in the angiotensin II mouse model, potentially via attenuation of cytokine/chemokine synthesis and the reduction of matrix metalloproteinase activities.
Assuntos
Aneurisma da Aorta Abdominal/fisiopatologia , Receptores de Prostaglandina E Subtipo EP4/fisiologia , Adulto , Angiotensina II , Animais , Aorta/metabolismo , Aorta/patologia , Aneurisma da Aorta Abdominal/induzido quimicamente , Aneurisma da Aorta Abdominal/diagnóstico por imagem , Aneurisma da Aorta Abdominal/prevenção & controle , Ruptura Aórtica/prevenção & controle , Aterosclerose/patologia , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/metabolismo , Avaliação Pré-Clínica de Medicamentos/métodos , Feminino , Técnicas de Silenciamento de Genes , Humanos , Macrófagos/efeitos dos fármacos , Masculino , Camundongos , Camundongos Knockout , Pessoa de Meia-Idade , Naftalenos/farmacologia , Naftalenos/uso terapêutico , Fenilbutiratos/farmacologia , Fenilbutiratos/uso terapêutico , Receptores de Prostaglandina E Subtipo EP4/antagonistas & inibidores , Receptores de Prostaglandina E Subtipo EP4/deficiência , Receptores de Prostaglandina E Subtipo EP4/metabolismo , Transdução de Sinais/fisiologia , UltrassonografiaRESUMO
Cysteinyl leukotrienes (CysLTs) are potent inflammatory mediators that predominantly exert their effects by binding to cysteinyl leukotriene receptors of the G protein-coupled receptor family. CysLT receptor 2 (CysLT(2)R), expressed in endothelial cells of some vascular beds, has been implicated in a variety of cardiovascular functions. Endothelium-specific overexpression of human CysLT(2)R in transgenic mice (hEC-CysLT(2)R) greatly increases myocardial infarction damage. Investigation of this receptor, however, has been hindered by the lack of selective pharmacological antagonists. Here, we describe the characterization of 3-(((3-carboxycyclohexyl)amino)carbonyl)-4-(3-(4-(4-phenoxybutoxy)phenyl)-propoxy)benzoic acid (BayCysLT(2)) and explore the selective effects of this compound in attenuating myocardial ischemia/reperfusion damage and vascular leakage. Using a recently developed ß-galactosidase-ß-arrestin complementation assay for CysLT(2)R activity (Mol Pharmacol 79:270-278, 2011), we determined BayCysLT(2) to be â¼20-fold more potent than the nonselective dual CysLT receptor 1 (CysLT(1)R)/CysLT(2)R antagonist 4-(((1R,2E,4E,6Z,9Z)-1-((1S)-4-carboxy-1-hydroxybutyl)-2,4,6,9-pentadecatetraen-1-yl)thio)benzoic acid (Bay-u9773) (IC(50) 274 nM versus 4.6 µM, respectively). Intracellular calcium mobilization in response to cysteinyl leukotriene administration showed that BayCysLT(2) was >500-fold more selective for CysLT(2)R compared with CysLT(1)R. Intraperitoneal injection of BayCysLT(2) in mice significantly attenuated leukotriene D(4)-induced Evans blue dye leakage in the murine ear vasculature. BayCysLT(2) administration either before or after ischemia/reperfusion attenuated the aforementioned increased myocardial infarction damage in hEC-CysLT(2)R mice. Finally, decreased neutrophil infiltration and leukocyte adhesion molecule mRNA expression were observed in mice treated with antagonist compared with untreated controls. In conclusion, we present the characterization of a potent and selective antagonist for CysLT(2)R that is useful for discerning biological activities of this receptor.
Assuntos
Permeabilidade Capilar/efeitos dos fármacos , Ácidos Cicloexanocarboxílicos/farmacologia , Antagonistas de Leucotrienos/farmacologia , Leucotrieno D4/antagonistas & inibidores , Infarto do Miocárdio/tratamento farmacológico , Traumatismo por Reperfusão Miocárdica/tratamento farmacológico , Ácidos Ftálicos/farmacologia , Receptores de Leucotrienos/metabolismo , SRS-A/análogos & derivados , Animais , Arrestinas/análise , Modelos Animais de Doenças , Avaliação Pré-Clínica de Medicamentos , Orelha/irrigação sanguínea , Humanos , Camundongos , Camundongos Transgênicos , Infarto do Miocárdio/metabolismo , Traumatismo por Reperfusão Miocárdica/fisiopatologia , Miocárdio/patologia , Miócitos Cardíacos/efeitos dos fármacos , Peroxidase/metabolismo , SRS-A/farmacologia , beta-Arrestinas , beta-Galactosidase/metabolismoRESUMO
INTRODUCTION: Analysis of nonlinear heart rate (HR) dynamics may provide greater insight into neurocardiac influences during exercise and disease than traditional HR variability. However, the physiological basis of nonlinear HR dynamics has not been investigated in individuals with spinal cord injury (SCI). The purpose of this study was to compare the effects of autonomic blockade in SCI and able-bodied participants. METHODS: Five participants (42 +/- 13 years) with SCI (C4-C7, AIS B-D, 13 +/- 13 years post-injury) and four able-bodied controls (33 +/- 8 years) underwent beta(1)-adrenergic and vagal blockade in the supine and cardiovascular stress positions. Cardiovascular stress consisted of 40 degrees tilt plus sustained isometric jaw contraction and cold water submersion of the right hand. RESULTS: In both SCI and able-bodied participants, vagal blockade significantly increased HR (p < 0.05) and resulted in significant reductions in sample entropy and correlation dimension in the supine and cardiovascular stress positions (p < 0.05). During the cardiovascular stress position, baseline sample entropy (p < 0.05) and correlation dimension (p < 0.05) were lower in participants with SCI. Nonlinear measures were also significantly correlated with HR (p < 0.05). CONCLUSION: The results suggest that vagal modulations are a primary modulator of nonlinear HR signals in both SCI and able-bodied participants, while the role of the beta(1)-adrenergic system remains less defined. Further study is required to elucidate the role of the autonomic nervous system in nonlinear HR dynamics in both SCI and able-bodied populations.
Assuntos
Sistema Nervoso Autônomo/fisiopatologia , Sistema Cardiovascular/fisiopatologia , Frequência Cardíaca/fisiologia , Adulto , Feminino , Cardiopatias/complicações , Humanos , Masculino , Pessoa de Meia-Idade , Padrões de Referência , Traumatismos da Medula Espinal/complicações , Estresse FisiológicoRESUMO
Arginase-1 catalyzes the conversion of arginine to ornithine and urea, which is the final step of the urea cycle used to remove excess ammonia from the body. Arginase-1 deficiency leads to hyperargininemia in mice and man with severe lethal consequences in the former and progressive neurological impairment to varying degrees in the latter. In a tamoxifen-induced arginase-1 deficient mouse model, mice succumb to the enzyme deficiency within 2 weeks after inducing the knockout and retain <2 % enzyme in the liver. Standard clinical care regimens for arginase-1 deficiency (low-protein diet, the nitrogen-scavenging drug sodium phenylbutyrate, ornithine supplementation) either failed to extend lifespan (ornithine) or only minimally prolonged lifespan (maximum 8 days with low-protein diet and drug). A conditional, tamoxifen-inducible arginase-1 transgenic mouse strain expressing the enzyme from the Rosa26 locus modestly extended lifespan of neonatal mice, but not that of 4-week old mice, when crossed to the inducible arginase-1 knockout mouse strain. Delivery of an arginase-1/enhanced green fluorescent fusion construct by adeno-associated viral delivery (rh10 serotype with a strong cytomegalovirus-chicken ß-actin hybrid promoter) rescued about 30% of male mice with lifespan prolongation to at least 6 months, extensive hepatic expression and restoration of significant enzyme activity in liver. In contrast, a vector of the AAV8 serotype driven by the thyroxine-binding globulin promoter led to weaker liver expression and did not rescue arginase-1 deficient mice to any great extent. Since the induced arginase-1 deficient mouse model displays a much more severe phenotype when compared to human arginase-1 deficiency, these studies reveal that it may be feasible with gene therapy strategies to correct the various manifestations of the disorder and they provide optimism for future clinical studies.
Assuntos
Arginase/genética , Animais , Arginase/metabolismo , Dependovirus/genética , Dieta com Restrição de Proteínas , Suplementos Nutricionais , Feminino , Expressão Gênica , Marcação de Genes , Genes Letais , Genes Reporter , Loci Gênicos , Vetores Genéticos/genética , Longevidade , Masculino , Camundongos , Camundongos Knockout , Ornitina/administração & dosagem , Ornitina/sangue , Fenótipo , RNA não Traduzido/genética , Transdução Genética , TransgenesRESUMO
Arginase deficiency is a rare autosomal recessive disorder resulting from a loss of the liver arginase isoform, arginase 1 (ARG1), which is the final step in the urea cycle for detoxifying ammonia. ARG1 deficiency leads to hyperargininemia, characterized by progressive neurological impairment, persistent growth retardation and infrequent episodes of hyperammonemia. Using the Cre/loxP-directed conditional gene knockout system, we generated an inducible Arg1-deficient mouse model by crossing "floxed" Arg1 mice with CreER(T2) mice. The resulting mice (Arg-Cre) die about two weeks after tamoxifen administration regardless of the starting age of inducing the knockout. These treated mice were nearly devoid of Arg1 mRNA, protein and liver arginase activity, and exhibited symptoms of hyperammonemia. Plasma amino acid analysis revealed pronounced hyperargininemia and significant alterations in amino acid and guanidino compound metabolism, including increased citrulline and guanidinoacetic acid. Despite no alteration in ornithine levels, concentrations of other amino acids such as proline and the branched-chain amino acids were reduced. In summary, we have generated and characterized an inducible Arg1-deficient mouse model exhibiting several pathologic manifestations of hyperargininemia. This model should prove useful for exploring potential treatment options of ARG1 deficiency.
Assuntos
Aminoácidos/metabolismo , Arginase/metabolismo , Hiperargininemia/enzimologia , Hiperargininemia/metabolismo , Aminoácidos de Cadeia Ramificada/metabolismo , Animais , Arginase/genética , Feminino , Hiperargininemia/genética , Masculino , Camundongos , Camundongos KnockoutRESUMO
The 5-lipoxygenase (5-LO) catalyzed formation of leukotriene (LT) lipid mediators is a pathway contributing to inflammatory events in asthma and more recently has been associated with cardiovascular disease. However, the relative impact of this pathway in atherogenesis has been controversial and a variety of mixed results reported. The goal of these studies was to assess the importance of the 5-LO/LT pathway in mice with either genetic (5-LO(-/-)) or pharmacological (L-739,010) inhibition of the 5-LO pathway on an apolipoprotein E deficient (apoE(-/-)) background when subjected to either an 8-week (Paigen) or 6 months (Western) atherosclerotic diet regimen. Atherosclerotic lesion analysis at the aortic root, brachiocephalic artery and throughout the whole aorta by en face Sudan IV staining was determined, as well as blood lipid levels. Ex vivo calcium ionophore-stimulation of whole blood demonstrated a significant reduction in the capacity to form LTB(4) in 5-LO(-/-) and drug-treated 5-LO(+/+) mice. Quantitative analysis of atherosclerotic lesions did not differ between groups at all three sites. Moreover, the composition of advanced lesions in the brachiocephalic arteries did not indicate altered plaque disruption as a result of 5-LO gene inactivation. These results do not support a role for the 5-LO/LT pathway in intermediate to advanced atherosclerotic lesion development in mice.