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1.
Hum Immunol ; 84(10): 515-524, 2023 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-37438188

RESUMEN

BACKGROUND: T helper 17 (Th17) cells produce IL-17A cytokine and can exacerbate autoimmune diseases and asthma. The ß2 adrenergic receptor is a g protein-coupled receptor that induces cAMP second messenger pathways. We tested the hypothesis that terbutaline, a ß2-adrenergic receptor-specific agonist, promotes IL-17 secretion by memory Th17 cells in a cAMP and PKA-dependent manner. METHODS: Venous peripheral blood mononuclear cells (PBMC) from healthy human participants were activated with anti-CD3 and anti-CD28 antibodies. Secreted IL-17A was measured by enzyme linked immunosorbent assay, intracellular IL-17A, and RORγ were measured using flow cytometry, and RORC by qPCR. Memory CD3+CD4+CD45RA-CD45RO+ T cells were obtained by immunomagnetic negative selection and activated with tri-antibody complex CD3/CD28/CD2. Secreted IL-17A, intracellular IL-17A, RORC were measured, and phosphorylated-serine133-CREB was measured by western blotting memory Th cells. RESULTS: Terbutaline increased IL-17A (p < 0.001), IL-17A+ cells (p < 0.05), and RORC in activated PBMC and memory Th cells. The PKA inhibitors H89 (p < 0.001) and Rp-cAMP (p < 0.01) abrogated the effects of terbutaline on IL-17A secretion in PBMC and memory T cells. Rolipram increased IL-17A (p < 0.01) to a similar extent as terbutaline. P-Ser133-CREB was increased by terbutaline (p < 0.05) in memory T cells. CONCLUSION: Terbutaline augments memory Th17 cells in lymphocytes from healthy participants. This could exacerbate autoimmune diseases or asthma, in cases where Th17 cells are considered to be pro-inflammatory.


Asunto(s)
Asma , Enfermedades Autoinmunes , Humanos , Agonistas Adrenérgicos/metabolismo , Agonistas Adrenérgicos/farmacología , Enfermedades Autoinmunes/metabolismo , Antígenos CD28/metabolismo , Proteínas Quinasas Dependientes de AMP Cíclico/metabolismo , Interleucina-17/metabolismo , Leucocitos Mononucleares/metabolismo , Receptores Adrenérgicos/metabolismo , Terbutalina/farmacología , Terbutalina/metabolismo , Células Th17
2.
FASEB J ; 35(12): e22033, 2021 12.
Artículo en Inglés | MEDLINE | ID: mdl-34739146

RESUMEN

The long-term use of adrenergic medication in treating various conditions, such as asthma, increases the chances of bone fracture. Dynamic mechanical loading at a specific time is a method for improving bone quality and promoting healing. Therefore, we hypothesized that precisely controlling the mechanical environment can contribute to the alleviation of the negative effects of chronic treatment with the common asthma drug terbutaline, which is a ß2-adrenergic receptor agonist that facilitates bone homeostasis and defect repair through its anabolic effect on osteogenic cells. Our in vitro results showed that terbutaline can directly inhibit osteogenesis by impairing osteogenic differentiation and mineralization. Chronic treatment in vivo was simulated by administering terbutaline to C57BL/6J mice for 4 weeks before bone defect surgery and mechanical loading. We utilized a stabilized tibial defect model, which allowed the application of anabolic mechanical loading. During homeostasis, chronic terbutaline treatment reduced the bone formation rate, the fracture toughness of long bones, and the concentrations of bone formation markers in the sera. During defect repair, terbutaline decreased the bone volume, type H vessel, and total blood vessel volume. Terbutaline treatment reduced the number of osteogenic cells. Periostin, which was secreted mainly by Prrx1+ osteoprogenitors and F4/80+ macrophages, was inhibited by treating the bone defect with terbutaline. Interestingly, controlled mechanical loading facilitated the recovery of bone volume and periostin expression and the number of osteogenic cells within the defect. In conclusion, mechanical loading can rescue negative effects on new bone accrual and repair induced by chronic terbutaline treatment.


Asunto(s)
Agonistas de Receptores Adrenérgicos beta 2/farmacología , Densidad Ósea , Regeneración Ósea , Diferenciación Celular , Receptores Adrenérgicos beta 2/química , Estrés Mecánico , Terbutalina/farmacología , Animales , Fenómenos Biomecánicos , Femenino , Homeostasis , Ratones , Ratones Endogámicos C57BL
3.
PLoS One ; 15(12): e0244253, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-33347508

RESUMEN

In order to elucidate involvement of cyclic AMP and intracellular Ca2+,[Ca2+]i, in the modulation of aqueous humour formation (AHF), we studied the effects of terbutaline, forskolin and 8-Br-cAMP in the isolated bovine eye. We also studied the interaction of cAMP on calcium signaling in cultured ciliary epithelial (CE) cells. Drug effects on AHF were measured by fluorescein dilution. Drug effects on [Ca2+]i were studied by the fura-2 fluorescence ratio technique. Terbutaline (100 nmol-100 M), forskolin (30 nM-100 M) or 8-Br-cAMP (100 nM- 10 µM), administered in the arterial perfusate produced significant reductions in AHF. The AH reducing effect of terbutaline was blocked by a selective inhibitor of protein kinase A (KT-5720). ATP (100 M) caused a rapid, transient (peak) increase in [Ca2+]i followed by a sustained plateau phase lasting more than 5 minutes. Preincubation of the cells (6 min) with terbutaline, forskolin or 8-Br-cAMP significantly reduced the peak calcium response to ATP. The sustained plateau phase of the response, on the other hand, was augmented by each of the agents. KT-5720 partially reversed the inhibitory effect of terbutaline on the peak and totally inhibited its effect on the plateau phase. These data indicate: (a) that AHF in the bovine eye can be manipulated through cyclic AMP, operating via protein kinase A, (b) that protein kinase A can affect [Ca2+]i homeostasis, (c) that calcium release from the intracellular store, not the entry, affects AHF, and (d) that interaction of [Ca2+]i with cAMP plays a role in modulating AH secretion.


Asunto(s)
Humor Acuoso/metabolismo , Secreciones Corporales/efectos de los fármacos , Calcio/metabolismo , Colforsina/farmacología , AMP Cíclico/farmacología , Terbutalina/farmacología , Animales , Humor Acuoso/efectos de los fármacos , Broncodilatadores/farmacología , Bovinos , Células Cultivadas , Proteínas Quinasas Dependientes de AMP Cíclico/metabolismo
4.
Exp Physiol ; 105(5): 886-892, 2020 05.
Artículo en Inglés | MEDLINE | ID: mdl-32170888

RESUMEN

NEW FINDINGS: What is the central question of this study? What is the role of ß2 -adrenergic receptor (ß2 AR) vasodilatation in older postmenopausal women as compared to premenopausal women and the role of nitric oxide (NO) in ß2 AR-mediated vasodilatation in both groups of women? What is the main finding and its importance? ß2 AR responsiveness is blunted in postmenopausal women compared to young premenopausal women. Additionally, NO may contribute to ß2 AR-mediated vasodilatation in young premenopausal women. ABSTRACT: ß2 -Adrenergic receptor (ß2 AR)-mediated vasodilatation, which is partially dependent on nitric oxide (NO) formation, is blunted in men at risk for developing hypertension. However, the role of ß2 AR vasodilatation in hypertension pathophysiology in ageing postmenopausal women is unclear. Therefore, the goals of this study were to determine if forearm vasodilatation to the selective ß2 AR agonist terbutaline is blunted in older postmenopausal women (59 ± 4 years) compared to young premenopausal women (27 ± 3 years) and to assess NO contribution to ß2 AR-mediated vasodilatation in both groups of women. Forearm blood flow (FBF) and forearm vascular conductance (FVC) were measured using venous occlusion plethysmography at baseline and during intra-arterial infusions of terbutaline at 0.1-2.0 µg (100 ml tissue)-1  min-1 with and without the NO synthase inhibitor l-NG -monomethylarginine (l-NMMA). Mean arterial pressure was significantly greater in postmenopausal women than in young women at baseline (P = 0.01). Baseline FBF and FVC did not differ between young and postmenopausal women (P > 0.05) and rose significantly within each group during terbutaline infusion (P < 0.05). There were significant group × dose interactions for FBF (P = 0.01) and FVC (P = 0.001), indicating vasodilator responses were lower in postmenopausal women. In young women, FVC response to the highest dose of terbutaline tended to be lower with l-NMMA co-infusion vs. without l-NMMA (P = 0.05). There were no significant decreases in FBF or FVC responses to terbutaline in postmenopausal women with l-NMMA co-infusion (P > 0.05 for all). These data suggest that ß2 AR responsiveness is blunted in postmenopausal women compared to young premenopausal women, and that NO may contribute to ß2 AR-mediated vasodilatation in young premenopausal women.


Asunto(s)
Agonistas Adrenérgicos/farmacología , Antebrazo/irrigación sanguínea , Terbutalina/farmacología , Vasodilatación , Adulto , Presión Arterial , Femenino , Humanos , Persona de Mediana Edad , Óxido Nítrico , Pletismografía , Posmenopausia , Premenopausia
5.
Pediatr Pulmonol ; 54(3): 280-288, 2019 03.
Artículo en Inglés | MEDLINE | ID: mdl-30644180

RESUMEN

Intrauterine inflammation generates inflammatory mediators that damage the developing bronchoalveolar epithelium, resulting in neonatal lung injury. Lung fluid transport disorders are the main reasons for the development of pulmonary edema, an important pathology of lung injury. Previous studies suggested that epithelial sodium channels (ENaCs) play an important role in lung fluid transport. Here, we investigated whether changes in the expression of ENaCs were observed when neonatal rat lung injury was induced by maternal exposure to endotoxin. We also examined the therapeutic effect of terbutaline nebulizer inhalation on this injury. The results showed that maternal exposure to endotoxin increased the levels of TNF-α and IL-1ß in bronchoalveolar lavage fluid, suppressed α-, ß-, γ-ENaC in the neonatal rat lung, and resulted in the formation of pulmonary edema on postnatal days 1 and 7. Terbutaline up-regulated the expression of ß- and γ-ENaC in the distal lung after 7 days of treatment. The potential signal molecules cAMP, PKA, and CREB expressions were increased after terbutaline treatment. In summary, maternal exposure to endotoxin decreased the expression of ENaCs in neonatal rats which, in turn, may exacerbate pulmonary edema. Inhalation of the ß2-adrenergic receptor agonist terbutaline improved lung liquid clearance. By increasing the expression of sodium ion channels, the effective removal of alveolar fluid provides a new way for the prevention and treatment of neonatal lung injury.


Asunto(s)
Agonistas Adrenérgicos beta/uso terapéutico , Endotoxinas/toxicidad , Canales Epiteliales de Sodio/metabolismo , Lesión Pulmonar/tratamiento farmacológico , Edema Pulmonar/tratamiento farmacológico , Terbutalina/uso terapéutico , Agonistas Adrenérgicos beta/farmacología , Animales , Animales Recién Nacidos , Transporte Biológico/efectos de los fármacos , Líquido del Lavado Bronquioalveolar/química , Líquido del Lavado Bronquioalveolar/inmunología , Femenino , Pulmón/efectos de los fármacos , Pulmón/metabolismo , Lesión Pulmonar/inducido químicamente , Lesión Pulmonar/inmunología , Lesión Pulmonar/metabolismo , Masculino , Intercambio Materno-Fetal , Embarazo , Edema Pulmonar/inducido químicamente , Edema Pulmonar/inmunología , Edema Pulmonar/metabolismo , Ratas Sprague-Dawley , Terbutalina/farmacología
6.
FEBS J ; 284(18): 3018-3028, 2017 09.
Artículo en Inglés | MEDLINE | ID: mdl-28710773

RESUMEN

Catecholamine hormones are powerful regulators of the immune system produced by the sympathetic nervous system (SNS). They regulate the adaptive immune system by altering T-cell differentiation into T helper (Th) 1 and Th2 cell subsets, but the effect on Th17 cells is not known. Th17 cells, defined, in part, by chemokine receptor CCR6 and cytokine interleukin (IL)-17A, are crucial for mediating certain pathogen-specific responses and are linked with several autoimmune diseases. We demonstrated that a proportion of human Th17 cells express beta 2-adrenergic receptor (ß2AR), a G protein-coupled receptor that responds to catecholamines. Activation of peripheral blood mononuclear cells, which were obtained from venous blood drawn from healthy volunteers, with anti-cluster of differentiation 3 (CD3) and anti-CD28 and with a ß2-agonist drug, terbutaline (TERB), augmented IL-17A levels (P < 0.01) in the majority of samples. TERB reduced interferon gamma (IFNγ) indicating that IL-17A and IFNγ are reciprocally regulated. Similar reciprocal regulation was observed with dbcAMP. Proliferation of Th cells was monitored by carboxyfluorescein diacetate N-succinimidyl ester labeling and flow cytometry with antibody staining for CD3 and CD4. TERB increased proliferation by a small but significant margin (P < 0.001). Next, Th17 cells (CD4+ CXCR3- CCR6+ ) were purified using an immunomagnetic positive selection kit, which removes all other mononuclear cells. TERB increased IL-17A from purified Th17 cells, which argues that TERB acts directly on Th17 cells. Thus, hormone signals from the SNS maintain a balance of Th cells subtypes through the ß2AR.


Asunto(s)
Agonistas de Receptores Adrenérgicos beta 2/farmacología , Interleucina-17/genética , Receptores Adrenérgicos beta 2/genética , Terbutalina/farmacología , Células TH1/efectos de los fármacos , Células Th17/efectos de los fármacos , Anticuerpos Monoclonales/farmacología , Bucladesina/inmunología , Antígenos CD28/antagonistas & inhibidores , Antígenos CD28/genética , Antígenos CD28/inmunología , Complejo CD3/genética , Complejo CD3/inmunología , Separación Celular , Regulación de la Expresión Génica , Humanos , Interferón gamma/genética , Interferón gamma/inmunología , Interleucina-17/inmunología , Cultivo Primario de Células , Receptores Adrenérgicos beta 2/inmunología , Transducción de Señal , Células TH1/citología , Células TH1/inmunología , Células Th17/citología , Células Th17/inmunología
7.
Pediatr Res ; 82(2): 349-355, 2017 08.
Artículo en Inglés | MEDLINE | ID: mdl-28288148

RESUMEN

Background: At birth, the release of surfactant from alveolar type II cells (ATIIs) is stimulated by increased activity of the beta-adrenergic/adenylyl cyclase/cyclic 3'-5' adenosine monophosphate-signaling cascade. Atrial natriuretic peptide (ANP) stimulates surfactant secretion through natriuretic peptide receptor A (NPR-A). ANP inhibits adenylyl cyclase activity through its binding to NPR-C. We wished to further understand the role of the NPR-C in perinatal transition. Methods: We studied ATII expression of NPR-C in fetal and newborn sheep using immunohistochemistry, and surfactant secretion in isolated ATIIs by measuring 3[H] choline release into the media. Results: ANP induced surfactant secretion, and, at higher doses, it inhibits the stimulatory effect of the secretagogue terbutaline. ATII NPR-C expression decreased significantly after birth. Premature delivery also markedly decreased ANP and NPR-C in ATIIs. Co-incubation of terbutaline (10-4 M) with ANP (10-6 M) significantly decreased 3[H] choline release from isolated newborn ATII cells when compared with terbutaline alone; this inhibitory effect was mimicked by the specific NPR-C agonist, C-ANP (10-10 M). Conclusion: ANP may act as an important epithelial-derived inhibitor of surfactant release in the fetal lung, and downregulation of ANP and NPR-C following birth may sensitize ATII cells to the effects of circulating catecholamines, thus facilitating surfactant secretion.


Asunto(s)
Pulmón/metabolismo , Péptido Natriurético Tipo-C/metabolismo , Ovinos/embriología , Animales , Animales Recién Nacidos , Inmunohistoquímica , Toxina del Pertussis/farmacología , Surfactantes Pulmonares/metabolismo , Terbutalina/farmacología
8.
Cytokine ; 94: 1-7, 2017 06.
Artículo en Inglés | MEDLINE | ID: mdl-28162907

RESUMEN

ß2-receptor agonists are used in the treatment of inflammatory obstructive lung diseases asthma and COPD as a symptomatic remedy, but they have been suggested to possess anti-inflammatory properties, also. ß2-receptor activation is considered to lead to the activation of ERK pathway through G-protein- and cAMP-independent mechanisms. In this study, we investigated the effects of ß2-receptor agonists salbutamol and terbutaline on the production of inflammatory factors in macrophages. We found that ß2-receptor agonists inhibited LPS-induced ERK phosphorylation and the production of MCP-1. A chemical cAMP analog 8-Br-cAMP also inhibited ERK phosphorylation and TNF and MCP-1 release. As expected, MAPK/ERK kinase (MEK)1/2 inhibitor PD0325901 inhibited ERK phosphorylation and suppressed both TNF and MCP-1 production. In conclusion, we suggest that ß2-receptor agonists salbutamol and terbutaline inhibit inflammatory gene expression partly by a mechanism dependent on cAMP leading to the inhibition of ERK signaling in macrophages. Observed anti-inflammatory effects of ß2-receptor agonists may contribute to the clinical effects of these drugs.


Asunto(s)
Agonistas de Receptores Adrenérgicos beta 2/farmacología , Albuterol/farmacología , Citocinas/biosíntesis , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Macrófagos/efectos de los fármacos , Terbutalina/farmacología , 8-Bromo Monofosfato de Adenosina Cíclica/análogos & derivados , 8-Bromo Monofosfato de Adenosina Cíclica/farmacología , Animales , Benzamidas/farmacología , Línea Celular , Quimiocina CCL2/metabolismo , AMP Cíclico/metabolismo , Citocinas/genética , Difenilamina/análogos & derivados , Difenilamina/farmacología , Factores Inmunológicos/farmacología , Mediciones Luminiscentes , Ratones , Fosforilación/efectos de los fármacos , Factor de Necrosis Tumoral alfa/metabolismo
9.
J Cell Physiol ; 232(2): 298-308, 2017 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-27403604

RESUMEN

Angiogenesis is associated with changes in endothelial cell (EC) proliferation and tube formation, controlled by extracellular receptor-activated kinase (ERK)/mitogen activated protein kinase (MAPK) and Akt signaling. Important regulators of these systems include hormones acting on G-protein-coupled receptors, such as beta 2-adrenoceptors (ß2-ARs). In central nervous system (CNS) trauma, the importance of ß2-AR modulation has been highlighted, although the effects on revascularization remain unclear. Vascular protection and revascularization are, however, key to support regeneration. We have investigated the angiogenic capacity of the specific ß2-AR agonist terbutaline on ECs derived from the CNS, namely bEnd.3-cells. As angiogenesis is a multistep process involving increased proliferation and tube formation of ECs, we investigated the effects of terbutaline on these processes. We show that terbutaline significantly induced bEnd.3 tube formation in a matrigel in vitro assay. Moreover, administration of specific inhibitors of ERK and Akt signaling both inhibited terbutaline-induced tube formation. The proliferation rate of the ECs was not affected. In order to investigate the general effects of terbutaline in an organotypic system, we have used the chick chorioallantoic membrane (CAM)-assay. Most importantly, terbutaline increased the number of blood vessels in this in ovo setting. Although we observed a positive trend, the systemic administration of terbutaline did not significantly improve the functional outcome, nor did it affect revascularization in our spinal cord injury model. In conclusion, these data indicate that terbutaline is promising to stimulate blood vessel formation, underscoring the importance of further research into the angiotherapeutic relevance of terbutaline and ß2-AR signaling after CNS-trauma. J. Cell. Physiol. 232: 298-308, 2017. © 2016 Wiley Periodicals, Inc.


Asunto(s)
Agonistas de Receptores Adrenérgicos beta 2/farmacología , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Neovascularización Fisiológica/efectos de los fármacos , Proteínas Proto-Oncogénicas c-akt/metabolismo , Receptores Adrenérgicos beta 2/metabolismo , Transducción de Señal/efectos de los fármacos , Terbutalina/farmacología , Animales , Línea Celular , Proliferación Celular/efectos de los fármacos , Pollos , Membrana Corioalantoides/efectos de los fármacos , Membrana Corioalantoides/metabolismo , Células Endoteliales/citología , Células Endoteliales/efectos de los fármacos , Células Endoteliales/metabolismo , Ratones , Modelos Biológicos , Regeneración/efectos de los fármacos , Traumatismos de la Médula Espinal/tratamiento farmacológico , Traumatismos de la Médula Espinal/patología , Terbutalina/uso terapéutico
10.
Scand J Med Sci Sports ; 27(7): 684-691, 2017 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-27060857

RESUMEN

While chronic systemic administration of glucocorticoids increases muscle Na+ ,K+ ATPase content, such effect is unexplored after therapeutic inhalation. We investigated the effect of therapeutic inhalation of the glucocorticoid budesonide on Na+ ,K+ ATPase content of skeletal muscle in men. Ten healthy trained subjects, aged 23 ± 4 years (mean ± 95% CI), participated in the study. Before and after 2 weeks of daily inhalation of budesonide (1.6 mg/day), a biopsy was taken from the vastus lateralis muscle for measurement of Na+ ,K+ ATPase content and blood samples were drawn for determination of plasma budesonide, cortisol, and K+ . Subjects' performance during cycling to fatigue at 90% of incremental peak power output (iPPO) was measured in response to 4 mg inhaled terbutaline to maximally stimulate Na+ ,K+ ATPase activity. Plasma concentrations of budesonide rose to 5.0 ± 1.6 nM with the intervention, whereas no changes were observed in plasma cortisol. Muscle Na+ ,K+ ATPase content increased (P ≤ 0.01) by 46 ± 34 pmol/(g wet wt) (17% increase) with the intervention. Cycling performance at 90% of iPPO did not change (P = 0.21) with the intervention (203 vs 214 s) in response to terbutaline. The present observations show that therapeutic inhalation of glucocorticoids increases muscle Na+ ,K+ ATPase content, but does not enhance high-intensity cycling endurance in response to terbutaline.


Asunto(s)
Budesonida/farmacología , Ejercicio Físico/fisiología , Resistencia Física/efectos de los fármacos , Músculo Cuádriceps/efectos de los fármacos , ATPasa Intercambiadora de Sodio-Potasio/metabolismo , Terbutalina/farmacología , Administración por Inhalación , Adulto , Biopsia , Budesonida/administración & dosificación , Budesonida/sangre , Humanos , Hidrocortisona/sangre , Estudios Longitudinales , Masculino , Potasio/sangre , Músculo Cuádriceps/enzimología , Terbutalina/administración & dosificación , Adulto Joven
11.
Am J Respir Cell Mol Biol ; 55(3): 395-406, 2016 09.
Artículo en Inglés | MEDLINE | ID: mdl-27064541

RESUMEN

Active ion transport by basolateral Na-K-ATPase (Na pump) creates an Na(+) gradient that drives fluid absorption across lung alveolar epithelium. The α1 and ß1 subunits are the most highly expressed Na pump subunits in alveolar epithelial cells (AEC). The specific contribution of the ß1 subunit and the relative contributions of alveolar epithelial type II (AT2) versus type I (AT1) cells to alveolar fluid clearance (AFC) were investigated using two cell type-specific mouse knockout lines in which the ß1 subunit was knocked out in either AT1 cells or both AT1 and AT2 cells. AFC was markedly decreased in both knockout lines, revealing, we believe for the first time, that AT1 cells play a major role in AFC and providing insights into AEC-specific roles in alveolar homeostasis. AEC monolayers derived from knockout mice demonstrated decreased short-circuit current and active Na(+) absorption, consistent with in vivo observations. Neither hyperoxia nor ventilator-induced lung injury increased wet-to-dry lung weight ratios in knockout lungs relative to control lungs. Knockout mice showed increases in Na pump ß3 subunit expression and ß2-adrenergic receptor expression. These results demonstrate a crucial role for the Na pump ß1 subunit in alveolar ion and fluid transport and indicate that both AT1 and AT2 cells make major contributions to these processes and to AFC. Furthermore, they support the feasibility of a general approach to altering alveolar epithelial function in a cell-specific manner that allows direct insights into AT1 versus AT2 cell-specific roles in the lung.


Asunto(s)
Células Epiteliales Alveolares/metabolismo , Líquidos Corporales/metabolismo , Absorción Fisiológica , Células Epiteliales Alveolares/patología , Amilorida/farmacología , Animales , Marcación de Gen , Hiperoxia/complicaciones , Hiperoxia/patología , Activación del Canal Iónico/efectos de los fármacos , Ratones Noqueados , Tamaño de los Órganos , Permeabilidad , Subunidades de Proteína/metabolismo , Edema Pulmonar/metabolismo , Edema Pulmonar/patología , Edema Pulmonar/fisiopatología , Receptores Adrenérgicos beta 2/metabolismo , Reproducibilidad de los Resultados , Sodio/metabolismo , Canales de Sodio/metabolismo , ATPasa Intercambiadora de Sodio-Potasio/metabolismo , Terbutalina/farmacología , Lesión Pulmonar Inducida por Ventilación Mecánica/complicaciones , Lesión Pulmonar Inducida por Ventilación Mecánica/patología , Lesión Pulmonar Inducida por Ventilación Mecánica/fisiopatología
12.
Chirality ; 28(4): 306-12, 2016 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-26969816

RESUMEN

In this study an enantioseparation method for rac-bambuterol (5-(2-(tert-butylamino)-1-hydroxyethyl)-1,3-phenylene bis(dimethylcarbamate)) via diastereoisomeric salt formation with o-chloromandelic acid was developed. The enantiomeric excess (ee) values and chemical purities of the desired products were confirmed by high-performance liquid chromatography (HPLC) using chiral stationary phase and reverse-phase HPLC analyses, respectively. The ee values and the chemical purities both exceeded 99%. Animal experiments showed that (R)-bambuterol was a potent inhibitor for histamine-induced asthma reactions. (S)-bambuterol was ineffective in relaxing the airways. Both enantiomers increased heart rates in beagles. Therefore, replacing rac-bambuterol with (R)-bambuterol could be beneficial for asthma patients.


Asunto(s)
Histamina/química , Ácidos Mandélicos/química , Terbutalina/análogos & derivados , Animales , Cromatografía Líquida de Alta Presión , Cobayas , Humanos , Estereoisomerismo , Terbutalina/química , Terbutalina/farmacología
13.
Lung ; 194(3): 401-8, 2016 06.
Artículo en Inglés | MEDLINE | ID: mdl-26989055

RESUMEN

INTRODUCTION: γ-amino butyric acid (GABA) is not only the major inhibitory neurotransmitter in the central nervous system (CNS), but it also plays an important role in the lung, mediating airway smooth muscle relaxation and mucus production. As kinases such as protein kinase A (PKA) are known to regulate the release and reuptake of GABA in the CNS by GABA transporters, we hypothesized that ß-agonists would affect GABA release from airway epithelial cells through activation of PKA. METHODS: C57/BL6 mice received a pretreatment of a ß-agonist or vehicle (PBS), followed by methacholine or PBS. Bronchoalveolar lavage (BAL) was collected and the amount of GABA was quantified using HPLC mass spectrometry. For in vitro studies, cultured BEAS-2B human airway epithelial cells were loaded with (3)H-GABA. (3)H-GABA released was measured during activation and inhibition of PKA and tyrosine kinase signaling pathways. RESULTS: ß-agonist pretreatment prior to methacholine challenge attenuated in vivo GABA release in mouse BAL and (3)H-GABA release from depolarized BEAS-2B cells. GABA release was also decreased in BEAS-2B cells by increases in cAMP but not by Epac or tyrosine kinase activation. CONCLUSION: ß-agonists decrease GABA release from airway epithelium through the activation of cAMP and PKA. This has important therapeutic implications as ß-agonists and GABA are important mediators of both mucus production and airway smooth muscle tone.


Asunto(s)
Agonistas Adrenérgicos beta/farmacología , Proteínas Quinasas Dependientes de AMP Cíclico/metabolismo , Células Epiteliales/metabolismo , Mucosa Respiratoria/metabolismo , Terbutalina/farmacología , Ácido gamma-Aminobutírico/metabolismo , Antagonistas Adrenérgicos beta/farmacología , Animales , Líquido del Lavado Bronquioalveolar/química , Línea Celular , Colforsina/farmacología , AMP Cíclico/metabolismo , Proteínas Quinasas Dependientes de AMP Cíclico/antagonistas & inhibidores , Activadores de Enzimas/farmacología , Glutamato Descarboxilasa/genética , Factores de Intercambio de Guanina Nucleótido/metabolismo , Humanos , Masculino , Cloruro de Metacolina/farmacología , Ratones , Ratones Endogámicos C57BL , Miocitos del Músculo Liso/metabolismo , Factor de Crecimiento Derivado de Plaquetas/farmacología , Propranolol/farmacología , ARN Mensajero/metabolismo , Proteínas Tirosina Quinasas Receptoras/antagonistas & inhibidores , Mucosa Respiratoria/citología , Rifabutina/análogos & derivados , Rifabutina/farmacología , Transducción de Señal/efectos de los fármacos , Ácido gamma-Aminobutírico/análisis
14.
PLoS One ; 11(2): e0148144, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-26849227

RESUMEN

Mitogen-activated protein kinase phosphatase 1 (MKP-1) expression is induced by inflammatory factors, and it is an endogenous suppressor of inflammatory response. MKP-1 expression is increased by PDE4 inhibitor rolipram suggesting that it is regulated by cAMP-enhancing compounds. Therefore, we investigated the effect of ß2-receptor agonists on MKP-1 expression and inflammatory response. We found that ß2-receptor agonists salbutamol and terbutaline, as well as 8-Br-cAMP, increased MKP-1 expression. Salbutamol and terbutaline also inhibited p38 MAPK phosphorylation and TNF production in J774 mouse macrophages. Interestingly, salbutamol suppressed carrageenan-induced paw inflammation in wild-type mice, but the effect was attenuated in MKP-1(-/-) mice. In conclusion, these data show that ß2-receptor agonists increase MKP-1 expression, which seems to mediate, at least partly, the observed anti-inflammatory effects of ß2-receptor agonists.


Asunto(s)
Agonistas de Receptores Adrenérgicos beta 2/farmacología , Albuterol/farmacología , Antiinflamatorios/farmacología , Fosfatasa 1 de Especificidad Dual/metabolismo , Terbutalina/farmacología , Animales , Línea Celular , Fosfatasa 1 de Especificidad Dual/genética , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Ratones , Ratones Endogámicos C57BL , Factor de Necrosis Tumoral alfa/biosíntesis
15.
Med Sci Sports Exerc ; 48(1): 39-48, 2016 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-26197029

RESUMEN

PURPOSE: The aim of the present study was to investigate the effects of the beta2-agonist terbutaline (TER) on power output and muscle metabolism during maximal sprint cycling. METHODS: In a randomized double-blind cross-over design, nine moderately trained men (VO2max = 4.6 ± 0.2 L · min(-1)) conducted a 10-s cycle sprint after inhalation of either 15 mg of TER or placebo (PLA). A muscle biopsy sample was collected before and <10 s after the sprint and was analyzed for metabolites. RESULTS: The mean power and peak power during the sprint were 8.3% ± 1.1% and 7.8% ± 2.5% higher (P < 0.05) with TER than with PLA, respectively. Moreover, the net rates of glycogenolysis (6.5 ± 0.8 vs 3.1 ± 0.7 mmol glucosyl units · kg dry weight(-1) · s(-1)) and glycolysis (2.4 ± 0.2 vs 1.6 ± 0.2 mmol glucosyl units · kg dry weight(-1) · s(-1)) were higher (P < 0.05) with TER than with PLA. After the sprint, adenosine triphosphate (ATP) was reduced with PLA (P < 0.05) but not with TER. During the sprint, there was no difference in the breakdown of phosphocreatine (PCr) between treatments. Estimated anaerobic ATP utilization was 9.2% ± 4.0% higher (P < 0.05) with TER than with PLA. After the sprint, ATP in Type II fibers was lowered (P < 0.05) by 25.7% ± 7.3% with PLA but was not reduced with TER. Before the sprint, PCr in Type II fibers was 24.5% ± 7.2% lower (P < 0.05) with TER than with PLA. With PLA, breakdown of PCr was 50.2% ± 24.8% higher (P < 0.05) in Type II fibers (vs Type I fibers), whereas no difference was observed between fiber types with TER. CONCLUSION: The present study shows that a TER-induced increase in power output is associated with increased rates of glycogenolysis and glycolysis in skeletal muscles. Furthermore, as TER counteracts a reduction in ATP in Type II fibers, TER may postpone fatigue development in these fibers.


Asunto(s)
Agonistas de Receptores Adrenérgicos beta 2/farmacología , Ciclismo/fisiología , Glucólisis/efectos de los fármacos , Músculo Esquelético/efectos de los fármacos , Terbutalina/farmacología , Adenosina Trifosfato/metabolismo , Estudios Cruzados , Método Doble Ciego , Metabolismo Energético , Humanos , Masculino , Fibras Musculares Esqueléticas/efectos de los fármacos , Fibras Musculares Esqueléticas/metabolismo , Músculo Esquelético/metabolismo , Fosfocreatina/metabolismo , Intercambio Gaseoso Pulmonar , Adulto Joven
16.
Am J Physiol Regul Integr Comp Physiol ; 308(7): R636-49, 2015 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-25632025

RESUMEN

In pulmonary epithelia, ß-adrenergic agonists regulate the membrane abundance of the epithelial sodium channel (ENaC) and, thereby, control the rate of transepithelial electrolyte absorption. This is a crucial regulatory mechanism for lung liquid clearance at birth and thereafter. This study investigated the influence of the gaseous signaling molecule hydrogen sulfide (H2S) on ß-adrenergic agonist-regulated pulmonary sodium and liquid absorption. Application of the H2S-liberating molecule Na2S (50 µM) to the alveolar compartment of rat lungs in situ decreased baseline liquid absorption and abrogated the stimulation of liquid absorption by the ß-adrenergic agonist terbutaline. There was no additional effect of Na2S over that of the ENaC inhibitor amiloride. In electrophysiological Ussing chamber experiments with native lung epithelia (Xenopus laevis), Na2S inhibited the stimulation of amiloride-sensitive current by terbutaline. ß-adrenergic agonists generally increase ENaC abundance by cAMP formation and activation of PKA. Activation of this pathway by forskolin and 3-isobutyl-1-methylxanthine increased amiloride-sensitive currents in H441 pulmonary epithelial cells. This effect was inhibited by Na2S in a dose-dependent manner (5-50 µM). Na2S had no effect on cellular ATP concentration, cAMP formation, and activation of PKA. By contrast, Na2S prevented the cAMP-induced increase in ENaC activity in the apical membrane of H441 cells. H441 cells expressed the H2S-generating enzymes cystathionine-ß-synthase, cystathionine-γ-lyase, and 3-mercaptopyruvate sulfurtransferase, and they produced H2S amounts within the employed concentration range. These data demonstrate that H2S prevents the stimulation of ENaC by cAMP/PKA and, thereby, inhibits the proabsorptive effect of ß-adrenergic agonists on lung liquid clearance.


Asunto(s)
Agonistas Adrenérgicos beta/farmacología , Células Epiteliales/efectos de los fármacos , Canales Epiteliales de Sodio/efectos de los fármacos , Sulfuro de Hidrógeno/metabolismo , Alveolos Pulmonares/efectos de los fármacos , Absorción a través del Sistema Respiratorio/efectos de los fármacos , Sodio/metabolismo , Sulfuros/farmacología , Terbutalina/farmacología , Animales , Línea Celular Tumoral , AMP Cíclico/metabolismo , Proteínas Quinasas Dependientes de AMP Cíclico/metabolismo , Cistationina betasintasa/genética , Cistationina betasintasa/metabolismo , Cistationina gamma-Liasa/genética , Cistationina gamma-Liasa/metabolismo , Relación Dosis-Respuesta a Droga , Activación Enzimática , Células Epiteliales/metabolismo , Canales Epiteliales de Sodio/metabolismo , Humanos , Masculino , Potenciales de la Membrana , Alveolos Pulmonares/metabolismo , ARN Mensajero/metabolismo , Ratas Wistar , Sulfurtransferasas/genética , Sulfurtransferasas/metabolismo , Factores de Tiempo , Xenopus laevis
17.
Acta Pharm Hung ; 84(3): 111-9, 2014.
Artículo en Húngaro | MEDLINE | ID: mdl-25470978

RESUMEN

Reactive oxygen intermediers (ROI) play a role in the signal transduction of beta-adrenergic receptors. We investigated whether an antioxidant (limonene) can reduce the beta-mimetic effect of terbutaline in beta-2-adrenergic receptor (ß2-AR)-regulated smooth muscles. Tissue samples were collected from nonpregnant (trachea) and 22-day-pregnant (myometrium and cervix) rats. Tissue contractility was investigated in an isolated organ bath. In separate groups of animals, the tracheal and uterine ß2-AR activities were upregulated by 17-beta-estradiol valerate (E2) and progesterone (P4), respectively. The total oxidant (TOS) and total antioxidant status (TAS) were also measured. The oxidative stress index (OSI) was defined as the ratio ofTOS and TAS. Terbutaline (10(-10) - 10(-5) M) decreased the spontaneous contractions in the nontreated and the P4-pretreated myometria. The concentration-response curves for terbutaline in the presence of 10-3 M limonene were shifted to the left, but the maximum inhibitory effect was unchanged. Terbutaline (10(-6) M) increased the cervical resistance both in the nontreated and in the P4-treated samples, while limonene reduced this action only in the P4-treated cervices. Terbutaline (10(-9) - 10(-4) M) reduced the tracheal tones both in the nontreated and in the E2-treated tissues, while limonene reduced these effects. The OSI was highest in the trachea and lowest in the pregnant myometrium. Limonene has various influence on terbutaline induced effects in certain tissues. Higher OSI value means, that the antioxidants have greater role in the beta-adrenergic signalmechanism. We assume that the significance of ROI in the signalling process of the ß2-ARs are divergent in the various tissues. Our results suggest that the antiasthmatic effect of beta-mimetics may worsen during parallel limonene administration.


Asunto(s)
Antagonistas Adrenérgicos beta/farmacología , Antioxidantes/farmacología , Ciclohexenos/farmacología , Contracción Muscular/efectos de los fármacos , Músculo Liso/efectos de los fármacos , Músculo Liso/metabolismo , Estrés Oxidativo , Receptores Adrenérgicos beta 2/metabolismo , Transducción de Señal/efectos de los fármacos , Terbutalina/antagonistas & inhibidores , Terpenos/farmacología , Animales , Estradiol/análogos & derivados , Estradiol/farmacología , Femenino , Limoneno , Estrés Oxidativo/efectos de los fármacos , Valor Predictivo de las Pruebas , Embarazo , Progesterona/farmacología , Ratas , Terbutalina/farmacología , Tráquea/metabolismo , Regulación hacia Arriba , Contracción Uterina/efectos de los fármacos
18.
Life Sci ; 105(1-2): 48-55, 2014 Jun 06.
Artículo en Inglés | MEDLINE | ID: mdl-24780318

RESUMEN

AIMS: Reactive oxygen species play a role in the signal transduction of beta-adrenergic receptors. We investigated whether an antioxidant (tocopherol) can reduce the effect of terbutaline in beta-2-adrenergic receptor (ß2-AR)-regulated smooth muscles. MAIN METHODS: Contractility of the tissues from nonpregnant (trachea) and 22-day-pregnant (myometrium and cervix) rats was investigated in an isolated organ bath. The tracheal and uterine ß2-AR expressions were increased by 17-beta-estradiol valerate (E2) and progesterone (P4), respectively. The accumulation of cyclic-AMP (cAMP), and the total oxidant (TOS) and total antioxidant status (TAS) were also measured. The oxidative stress index (OSI) was defined as the ratio of TOS and TAS. KEY FINDINGS: Terbutaline (10(-10)-10(-5)M) decreased the contractions in the nontreated and the P4-pretreated myometria, but tocopherol (10(-7)M) did not alter these actions. Terbutaline (10(-6)M) increased the cervical resistance both in the nontreated and in the P4-treated samples, while tocopherol reduced this action only in the P4-treated cervices. Terbutaline (10(-9)-10(-4)M) reduced the tracheal tones both in the nontreated and in the E2-treated tissues, while tocopherol reduced these effects. The changes in the intracellular cAMP levels of the tissues were in harmony with the isolated organ results. The OSI was highest in the trachea and lowest in the pregnant myometrium. SIGNIFICANCE: A higher OSI is linked to a higher tocopherol sensitivity of beta-mimetic-induced relaxation. Our results suggest that the antiasthmatic effect of beta-mimetics may worsen, while their tocolytic effect may remain unchanged during parallel tocopherol administration.


Asunto(s)
Músculo Liso/metabolismo , Estrés Oxidativo/fisiología , Fenómenos Fisiológicos Reproductivos/efectos de los fármacos , Sistema Respiratorio/efectos de los fármacos , Terbutalina/antagonistas & inhibidores , Tocoferoles/farmacología , Análisis de Varianza , Animales , Antioxidantes/metabolismo , Western Blotting , AMP Cíclico/metabolismo , Femenino , Contracción Muscular/efectos de los fármacos , Contracción Muscular/fisiología , Músculo Liso/efectos de los fármacos , Oxidantes/metabolismo , Estrés Oxidativo/efectos de los fármacos , Embarazo , Ratas , Ratas Sprague-Dawley , Especies Reactivas de Oxígeno/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Receptores Adrenérgicos beta 2/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Terbutalina/farmacología
19.
Clin Dev Immunol ; 2013: 764395, 2013.
Artículo en Inglés | MEDLINE | ID: mdl-24194774

RESUMEN

Adjuvant-induced arthritic (AA) differentially affects norepinephrine concentrations in immune organs, and in vivo ß-adrenergic receptor (ß-AR) agonist treatment distinctly regulates ex vivo cytokine profiles in different immune organs. We examined the contribution of altered ß-AR functioning in AA to understand these disparate findings. Twenty-one or 28 days after disease induction, we examined ß2-AR expression in spleen and draining lymph nodes (DLNs) for the arthritic limbs using radioligand binding and western blots and splenocyte ß-AR-stimulated cAMP production using enzyme-linked immunoassay (EIA). During severe disease, ß-AR agonists failed to induce splenocyte cAMP production, and ß-AR affinity and density declined, indicating receptor desensitization and downregulation. Splenocyte ß2-AR phosphorylation (pß2-AR) by protein kinase A (pß2-AR(PKA)) decreased in severe disease, and pß2-AR by G protein-coupled receptor kinases (pß2-AR(GRK)) increased in chronic disease. Conversely, in DLN cells, pß2-AR(PKA) rose during severe disease, but fell during chronic disease, and pß2-AR(GRK) increased during both disease stages. A similar pß2-AR pattern in DLN cells with the mycobacterial cell wall component of complete Freund's adjuvant suggests that pattern recognition receptors (i.e., toll-like receptors) are important for DLN pß2-AR patterns. Collectively, our findings indicate lymphoid organ- and disease stage-specific sympathetic dysregulation, possibly explaining immune compartment-specific differences in ß2-AR-mediated regulation of cytokine production in AA and rheumatoid arthritis.


Asunto(s)
Artritis Experimental/inmunología , Artritis Experimental/metabolismo , Receptores Adrenérgicos beta 2/metabolismo , Transducción de Señal , Sistema Nervioso Simpático/metabolismo , Agonistas de Receptores Adrenérgicos beta 2/administración & dosificación , Agonistas de Receptores Adrenérgicos beta 2/farmacología , Animales , Artritis Experimental/tratamiento farmacológico , Artritis Experimental/genética , AMP Cíclico/biosíntesis , Proteínas Quinasas Dependientes de AMP Cíclico/metabolismo , Quinasas de Receptores Acoplados a Proteína-G/metabolismo , Expresión Génica , Interferón gamma/biosíntesis , Ganglios Linfáticos/efectos de los fármacos , Ganglios Linfáticos/inmunología , Ganglios Linfáticos/metabolismo , Masculino , Fosforilación , Unión Proteica , Ratas , Receptores Adrenérgicos beta 2/genética , Índice de Severidad de la Enfermedad , Bazo/efectos de los fármacos , Bazo/inmunología , Bazo/metabolismo , Sistema Nervioso Simpático/fisiopatología , Terbutalina/administración & dosificación , Terbutalina/farmacología
20.
Shock ; 40(5): 430-6, 2013 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-24088990

RESUMEN

Aquaporin 1 (AQP1) and AQP5 expression may impact on key mechanisms in sepsis. However, it is unclear whether these AQPs are expressed to an equal extent or regulated differentially. Accordingly, we investigated the time-dependent expression of AQP1 and AQP5 following stimulation with lipopolysaccharide (LPS) in cultured human THP-1 cells and in the lungs of mice injected with LPS. Furthermore, we tested the hypothesis that the ß2 adrenoreceptor agonist terbutaline or its downstream effector cyclic adenosine monophosphate (cAMP) mitigates LPS-evoked changes of AQP expression. THP-1 cells were stimulated with either LPS (1 µg/mL; serotype O127:B8), 8-Br-cAMP (1 mM), or both, and RNA and protein were extracted at baseline and after 2, 6, and 24 h. C57BL/6 mice that received LPS (20 mg/kg i.p.), terbutaline (2.5 mg/kg), or both were killed 8 h later, and lungs were excised for RNA extraction and lung wet weight determination. Real-time polymerase chain reaction and Western blot analysis show that LPS increased AQP1 (3 h, P < 0.0001) but not AQP5 mRNA and protein expression in THP-1 cells. cAMP increased AQP1 (6 h, P < 0.0001) but not AQP5 mRNA and protein expression. Incubation with both substances accelerated the increase in AQP1 (2 h, P = 0.001) expression, whereas AQP5 expression decreased after 2 h but increased after 24 h (P = 0.0148). In mice lungs, LPS decreased AQP1 (P = 0.0082) but not AQP5 mRNA expression and increased lung wet weight. Terbutaline increased AQP1 mRNA expression twice (P = 0.0005) but not AQP5 mRNA expression. Terbutaline did neither abolish the LPS-induced decrease in AQP1 and AQP5 expression nor increase lung weight. Thus, AQP1 and AQP5 expression is differentially regulated following exposure to LPS, the ß2 adrenoreceptor agonist terbutaline, and cAMP. Furthermore, neither terbutaline nor cAMP mitigated the LPS-evoked change of AQP1 and AQP5 expression.


Asunto(s)
Acuaporina 1/biosíntesis , Acuaporina 5/biosíntesis , Lipopolisacáridos/farmacología , Monocitos/efectos de los fármacos , Terbutalina/farmacología , Agonistas de Receptores Adrenérgicos beta 2/farmacología , Animales , Acuaporina 1/genética , Acuaporina 5/genética , Línea Celular Tumoral , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Pulmón/anatomía & histología , Pulmón/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Monocitos/metabolismo , Tamaño de los Órganos/efectos de los fármacos , ARN Mensajero/genética
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