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Isolating Bronchial Epithelial Cells from Resected Lung Tissue for Biobanking and Establishing Well-Differentiated Air-Liquid Interface Cultures
Published on: May 26, 2023
(Endo)cannabinoids mediate different Ca2+ entry mechanisms in human bronchial epithelial cells.
Effimia Gkoumassi1, Bart G J Dekkers, Melloney J Dröge
1Department of Molecular Pharmacology, University Centre for Pharmacy, University of Groningen, A. Deusinglaan 1, 9713 AV Groningen, The Netherlands, e.gkoumassi@gmail.com
Naunyn-Schmiedeberg'S Archives of Pharmacology
|March 4, 2009
Summary
Cannabinoid receptors in lung cells modulate calcium signaling differently. Virodhamine activates distinct calcium entry pathways at high concentrations, impacting cellular responses.
Area of Science:
- Cell Biology
- Pharmacology
- Respiratory Medicine
Background:
- Human bronchial epithelial cells (16HBE14o(-)) express CB(1) and CB(2) cannabinoid receptors.
- Activation of these receptors by cannabinoids inhibits adenylyl cyclase and interleukin-8 release.
Purpose of the Study:
- To analyze intracellular calcium ([Ca2+](i)) changes induced by Delta(9)-tetrahydrocannabinol (Delta(9)-THC), CP55,940, and virodhamine in 16HBE14o(-) cells.
- To elucidate the mechanisms of calcium mobilization and entry pathways activated by these compounds.
Main Methods:
- Measurement of intracellular calcium ([Ca2+](i)) in 16HBE14o(-) cells.
- Pharmacological characterization of calcium influx using specific inhibitors (Ni2+, capsazepine, ruthenium red, La3+, Gd3+) and receptor antagonists.
- Analysis of mRNA and protein expression for TRPC and TRPV channels.
- Assay of arachidonic acid release.
Main Results:
- Delta(9)-THC and CP55,940 induced calcium mobilization and capacitative calcium entry (CCE).
- Low concentrations of virodhamine mimicked CP55,940 effects, involving CCE possibly via TRPC1.
- High concentrations of virodhamine induced non-CCE (NCCE) calcium entry, sensitive to TRPV1/TRPV4 inhibitors and involving TRPV1, TRPV4, and TRPC6.
- Virodhamine also enhanced arachidonic acid release, independent of cannabinoid receptors but sensitive to phospholipase A(2) inhibition and capsazepine.
Conclusions:
- Cannabinoid receptor activation in bronchial epithelial cells triggers distinct calcium signaling pathways.
- Low virodhamine concentrations activate CCE, while high concentrations activate NCCE pathways.
- Arachidonic acid release may contribute to virodhamine-induced calcium changes via TRPV1 activation.
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