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A fixed 20:1 combination of cafedrine/theodrenaline increases cytosolic Ca(2+) concentration in human tracheal epithelial cells via ryanodine receptor-mediated Ca(2+) release
Mucociliary clearance is a pivotal physiological mechanism that protects the lung by cleaning the airways from pollution and colonization, thereby preventing infection. Ciliary function is influenced by various signal transduction cascades, and Ca(2+) represents a key second messenger. A fixed 20:1...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Nature Publishing Group UK
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10533847/ https://www.ncbi.nlm.nih.gov/pubmed/37758747 http://dx.doi.org/10.1038/s41598-023-43342-0 |
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author | Schmidt, Götz Rienas, Gerrit Müller, Sabrina Richter, Katrin Sander, Michael Koch, Christian Henrich, Michael |
author_facet | Schmidt, Götz Rienas, Gerrit Müller, Sabrina Richter, Katrin Sander, Michael Koch, Christian Henrich, Michael |
author_sort | Schmidt, Götz |
collection | PubMed |
description | Mucociliary clearance is a pivotal physiological mechanism that protects the lung by cleaning the airways from pollution and colonization, thereby preventing infection. Ciliary function is influenced by various signal transduction cascades, and Ca(2+) represents a key second messenger. A fixed 20:1 combination of cafedrine and theodrenaline has been widely used to treat perioperative hypotension and emergency hypotensive states since the 1960s; however, its effect on the intracellular Ca(2+) concentration ([Ca(2+)](i)) of respiratory epithelium remains unknown. Therefore, human tracheal epithelial cells were exposed to the clinically applied 20:1 mixture of cafedrine/theodrenaline and the individual substances separately. [Ca(2+)](i) was assessed by FURA-2 340/380 fluorescence ratio. Pharmacological inhibitors were applied to elucidate relevant signal transduction cascades, and reverse transcription polymerase chain reaction (RT-PCR) was performed on murine tracheal epithelium to analyze ryanodine receptor (RyR) subtype expression. All three pharmacological preparations instantaneously induced a steep increase in [Ca(2+)](i) that quickly returned to its baseline value despite the persistence of each substance. Peak [Ca(2+)](i) following the administration of 20:1 cafedrine/theodrenaline, cafedrine alone, and theodrenaline alone increased in a dose-dependent manner, with median effective concentrations of 0.35 mM (7.32 mM cafedrine and 0.35 mM theodrenaline), 3.14 mM, and 3.45 mM, respectively. When extracellular Ca(2+) influx was inhibited using a Ca(2+)-free buffer solution, the peak [Ca(2+)](i) following the administration of cafedrine alone and theodrenaline alone were reduced but not abolished. No alteration in [Ca(2+)](i) compared with baseline [Ca(2+)](i) was observed during β-adrenergic receptor inhibition. Depletion of caffeine-sensitive stores and inhibition of RyR, but not IP(3) receptors, completely abolished any increase in [Ca(2+)](i). However, [Ca(2+)](i) still increased following the depletion of mitochondrial Ca(2+) stores using 2,4-dinitrophenol. RT-PCR revealed RyR-2 and RyR-3 expression on murine tracheal epithelium. Although our experiments showed that cafedrine/theodrenaline, cafedrine alone, or theodrenaline alone release Ca(2+) from intracellular stores through mechanisms that are exclusively triggered by β-adrenergic receptor stimulation, which most probably lead to RyR activation, clinical plasma concentrations are considerably lower than those used in our experiments to elicit an increase in [Ca(2+)](i); therefore, further studies are needed to evaluate the ability of cafedrine/theodrenaline to alter mucociliary clearance in clinical practice. |
format | Online Article Text |
id | pubmed-10533847 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-105338472023-09-29 A fixed 20:1 combination of cafedrine/theodrenaline increases cytosolic Ca(2+) concentration in human tracheal epithelial cells via ryanodine receptor-mediated Ca(2+) release Schmidt, Götz Rienas, Gerrit Müller, Sabrina Richter, Katrin Sander, Michael Koch, Christian Henrich, Michael Sci Rep Article Mucociliary clearance is a pivotal physiological mechanism that protects the lung by cleaning the airways from pollution and colonization, thereby preventing infection. Ciliary function is influenced by various signal transduction cascades, and Ca(2+) represents a key second messenger. A fixed 20:1 combination of cafedrine and theodrenaline has been widely used to treat perioperative hypotension and emergency hypotensive states since the 1960s; however, its effect on the intracellular Ca(2+) concentration ([Ca(2+)](i)) of respiratory epithelium remains unknown. Therefore, human tracheal epithelial cells were exposed to the clinically applied 20:1 mixture of cafedrine/theodrenaline and the individual substances separately. [Ca(2+)](i) was assessed by FURA-2 340/380 fluorescence ratio. Pharmacological inhibitors were applied to elucidate relevant signal transduction cascades, and reverse transcription polymerase chain reaction (RT-PCR) was performed on murine tracheal epithelium to analyze ryanodine receptor (RyR) subtype expression. All three pharmacological preparations instantaneously induced a steep increase in [Ca(2+)](i) that quickly returned to its baseline value despite the persistence of each substance. Peak [Ca(2+)](i) following the administration of 20:1 cafedrine/theodrenaline, cafedrine alone, and theodrenaline alone increased in a dose-dependent manner, with median effective concentrations of 0.35 mM (7.32 mM cafedrine and 0.35 mM theodrenaline), 3.14 mM, and 3.45 mM, respectively. When extracellular Ca(2+) influx was inhibited using a Ca(2+)-free buffer solution, the peak [Ca(2+)](i) following the administration of cafedrine alone and theodrenaline alone were reduced but not abolished. No alteration in [Ca(2+)](i) compared with baseline [Ca(2+)](i) was observed during β-adrenergic receptor inhibition. Depletion of caffeine-sensitive stores and inhibition of RyR, but not IP(3) receptors, completely abolished any increase in [Ca(2+)](i). However, [Ca(2+)](i) still increased following the depletion of mitochondrial Ca(2+) stores using 2,4-dinitrophenol. RT-PCR revealed RyR-2 and RyR-3 expression on murine tracheal epithelium. Although our experiments showed that cafedrine/theodrenaline, cafedrine alone, or theodrenaline alone release Ca(2+) from intracellular stores through mechanisms that are exclusively triggered by β-adrenergic receptor stimulation, which most probably lead to RyR activation, clinical plasma concentrations are considerably lower than those used in our experiments to elicit an increase in [Ca(2+)](i); therefore, further studies are needed to evaluate the ability of cafedrine/theodrenaline to alter mucociliary clearance in clinical practice. Nature Publishing Group UK 2023-09-27 /pmc/articles/PMC10533847/ /pubmed/37758747 http://dx.doi.org/10.1038/s41598-023-43342-0 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Schmidt, Götz Rienas, Gerrit Müller, Sabrina Richter, Katrin Sander, Michael Koch, Christian Henrich, Michael A fixed 20:1 combination of cafedrine/theodrenaline increases cytosolic Ca(2+) concentration in human tracheal epithelial cells via ryanodine receptor-mediated Ca(2+) release |
title | A fixed 20:1 combination of cafedrine/theodrenaline increases cytosolic Ca(2+) concentration in human tracheal epithelial cells via ryanodine receptor-mediated Ca(2+) release |
title_full | A fixed 20:1 combination of cafedrine/theodrenaline increases cytosolic Ca(2+) concentration in human tracheal epithelial cells via ryanodine receptor-mediated Ca(2+) release |
title_fullStr | A fixed 20:1 combination of cafedrine/theodrenaline increases cytosolic Ca(2+) concentration in human tracheal epithelial cells via ryanodine receptor-mediated Ca(2+) release |
title_full_unstemmed | A fixed 20:1 combination of cafedrine/theodrenaline increases cytosolic Ca(2+) concentration in human tracheal epithelial cells via ryanodine receptor-mediated Ca(2+) release |
title_short | A fixed 20:1 combination of cafedrine/theodrenaline increases cytosolic Ca(2+) concentration in human tracheal epithelial cells via ryanodine receptor-mediated Ca(2+) release |
title_sort | fixed 20:1 combination of cafedrine/theodrenaline increases cytosolic ca(2+) concentration in human tracheal epithelial cells via ryanodine receptor-mediated ca(2+) release |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10533847/ https://www.ncbi.nlm.nih.gov/pubmed/37758747 http://dx.doi.org/10.1038/s41598-023-43342-0 |
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