Cargando…
Dissociations in the Effects of β(2)-Adrenergic Receptor Agonists on cAMP Formation and Superoxide Production in Human Neutrophils: Support for the Concept of Functional Selectivity
In neutrophils, activation of the β(2)-adrenergic receptor (β(2)AR), a G(s)-coupled receptor, inhibits inflammatory responses, which could be therapeutically exploited. The aim of this study was to evaluate the effects of various β(2)AR ligands on adenosine-3′,5′-cyclic monophosphate (cAMP) accumula...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2013
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3669315/ https://www.ncbi.nlm.nih.gov/pubmed/23741338 http://dx.doi.org/10.1371/journal.pone.0064556 |
_version_ | 1782271731072761856 |
---|---|
author | Brunskole Hummel, Irena Reinartz, Michael T. Kälble, Solveig Burhenne, Heike Schwede, Frank Buschauer, Armin Seifert, Roland |
author_facet | Brunskole Hummel, Irena Reinartz, Michael T. Kälble, Solveig Burhenne, Heike Schwede, Frank Buschauer, Armin Seifert, Roland |
author_sort | Brunskole Hummel, Irena |
collection | PubMed |
description | In neutrophils, activation of the β(2)-adrenergic receptor (β(2)AR), a G(s)-coupled receptor, inhibits inflammatory responses, which could be therapeutically exploited. The aim of this study was to evaluate the effects of various β(2)AR ligands on adenosine-3′,5′-cyclic monophosphate (cAMP) accumulation and N-formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP)-induced superoxide anion (O(2) (•−)) production in human neutrophils and to probe the concept of ligand-specific receptor conformations (also referred to as functional selectivity or biased signaling) in a native cell system. This is an important question because so far, evidence for functional selectivity has been predominantly obtained with recombinant systems, due to the inherent difficulties to genetically manipulate human native cells. cAMP concentration was determined by HPLC/tandem mass spectrometry, and O(2) (•−) formation was assessed by superoxide dismutase-inhibitable reduction of ferricytochrome c. β(2)AR agonists were generally more potent in inhibiting fMLP-induced O(2) (•−) production than in stimulating cAMP accumulation. (−)-Ephedrine and dichloroisoproterenol were devoid of any agonistic activity in the cAMP assay, but partially inhibited fMLP-induced O(2) (•−) production. Moreover, (−)-adrenaline was equi-efficacious in both assays whereas the efficacy of salbutamol was more than two-fold higher in the O(2) (•−) assay. Functional selectivity was visualized by deviations of ligand potencies and efficacies from linear correlations for various parameters. We obtained no evidence for involvement of protein kinase A in the inhibition of fMLP-induced O(2) (•−) production after β(2)AR-stimulation although cAMP-increasing substances inhibited O(2) (•−) production. Taken together, our data corroborate the concept of ligand-specific receptor conformations with unique signaling capabilities in native human cells and suggest that the β(2)AR inhibits O(2) (•−) production in a cAMP-independent manner. |
format | Online Article Text |
id | pubmed-3669315 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-36693152013-06-05 Dissociations in the Effects of β(2)-Adrenergic Receptor Agonists on cAMP Formation and Superoxide Production in Human Neutrophils: Support for the Concept of Functional Selectivity Brunskole Hummel, Irena Reinartz, Michael T. Kälble, Solveig Burhenne, Heike Schwede, Frank Buschauer, Armin Seifert, Roland PLoS One Research Article In neutrophils, activation of the β(2)-adrenergic receptor (β(2)AR), a G(s)-coupled receptor, inhibits inflammatory responses, which could be therapeutically exploited. The aim of this study was to evaluate the effects of various β(2)AR ligands on adenosine-3′,5′-cyclic monophosphate (cAMP) accumulation and N-formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP)-induced superoxide anion (O(2) (•−)) production in human neutrophils and to probe the concept of ligand-specific receptor conformations (also referred to as functional selectivity or biased signaling) in a native cell system. This is an important question because so far, evidence for functional selectivity has been predominantly obtained with recombinant systems, due to the inherent difficulties to genetically manipulate human native cells. cAMP concentration was determined by HPLC/tandem mass spectrometry, and O(2) (•−) formation was assessed by superoxide dismutase-inhibitable reduction of ferricytochrome c. β(2)AR agonists were generally more potent in inhibiting fMLP-induced O(2) (•−) production than in stimulating cAMP accumulation. (−)-Ephedrine and dichloroisoproterenol were devoid of any agonistic activity in the cAMP assay, but partially inhibited fMLP-induced O(2) (•−) production. Moreover, (−)-adrenaline was equi-efficacious in both assays whereas the efficacy of salbutamol was more than two-fold higher in the O(2) (•−) assay. Functional selectivity was visualized by deviations of ligand potencies and efficacies from linear correlations for various parameters. We obtained no evidence for involvement of protein kinase A in the inhibition of fMLP-induced O(2) (•−) production after β(2)AR-stimulation although cAMP-increasing substances inhibited O(2) (•−) production. Taken together, our data corroborate the concept of ligand-specific receptor conformations with unique signaling capabilities in native human cells and suggest that the β(2)AR inhibits O(2) (•−) production in a cAMP-independent manner. Public Library of Science 2013-05-31 /pmc/articles/PMC3669315/ /pubmed/23741338 http://dx.doi.org/10.1371/journal.pone.0064556 Text en © 2013 Brunskole Hummel et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Brunskole Hummel, Irena Reinartz, Michael T. Kälble, Solveig Burhenne, Heike Schwede, Frank Buschauer, Armin Seifert, Roland Dissociations in the Effects of β(2)-Adrenergic Receptor Agonists on cAMP Formation and Superoxide Production in Human Neutrophils: Support for the Concept of Functional Selectivity |
title | Dissociations in the Effects of β(2)-Adrenergic Receptor Agonists on cAMP Formation and Superoxide Production in Human Neutrophils: Support for the Concept of Functional Selectivity |
title_full | Dissociations in the Effects of β(2)-Adrenergic Receptor Agonists on cAMP Formation and Superoxide Production in Human Neutrophils: Support for the Concept of Functional Selectivity |
title_fullStr | Dissociations in the Effects of β(2)-Adrenergic Receptor Agonists on cAMP Formation and Superoxide Production in Human Neutrophils: Support for the Concept of Functional Selectivity |
title_full_unstemmed | Dissociations in the Effects of β(2)-Adrenergic Receptor Agonists on cAMP Formation and Superoxide Production in Human Neutrophils: Support for the Concept of Functional Selectivity |
title_short | Dissociations in the Effects of β(2)-Adrenergic Receptor Agonists on cAMP Formation and Superoxide Production in Human Neutrophils: Support for the Concept of Functional Selectivity |
title_sort | dissociations in the effects of β(2)-adrenergic receptor agonists on camp formation and superoxide production in human neutrophils: support for the concept of functional selectivity |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3669315/ https://www.ncbi.nlm.nih.gov/pubmed/23741338 http://dx.doi.org/10.1371/journal.pone.0064556 |
work_keys_str_mv | AT brunskolehummelirena dissociationsintheeffectsofb2adrenergicreceptoragonistsoncampformationandsuperoxideproductioninhumanneutrophilssupportfortheconceptoffunctionalselectivity AT reinartzmichaelt dissociationsintheeffectsofb2adrenergicreceptoragonistsoncampformationandsuperoxideproductioninhumanneutrophilssupportfortheconceptoffunctionalselectivity AT kalblesolveig dissociationsintheeffectsofb2adrenergicreceptoragonistsoncampformationandsuperoxideproductioninhumanneutrophilssupportfortheconceptoffunctionalselectivity AT burhenneheike dissociationsintheeffectsofb2adrenergicreceptoragonistsoncampformationandsuperoxideproductioninhumanneutrophilssupportfortheconceptoffunctionalselectivity AT schwedefrank dissociationsintheeffectsofb2adrenergicreceptoragonistsoncampformationandsuperoxideproductioninhumanneutrophilssupportfortheconceptoffunctionalselectivity AT buschauerarmin dissociationsintheeffectsofb2adrenergicreceptoragonistsoncampformationandsuperoxideproductioninhumanneutrophilssupportfortheconceptoffunctionalselectivity AT seifertroland dissociationsintheeffectsofb2adrenergicreceptoragonistsoncampformationandsuperoxideproductioninhumanneutrophilssupportfortheconceptoffunctionalselectivity |