Pharmacological characterization of the calcium influx pathways involved in nitric oxide production by endothelial cells
ABSTRACT Objective: To characterize the calcium influx pathways implicated in the sustained elevation of endothelial intracellular calcium concentration, required for the synthesis and release of relaxing factors. Methods: We evaluated the effect of the newly synthesized pyrazole derivatives, desc...
| Main Authors: | , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Instituto Israelita de Ensino e Pesquisa Albert Einstein
|
| Series: | Einstein (São Paulo) |
| Subjects: | |
| Online Access: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1679-45082019000300204&lng=en&tlng=en |
| _version_ | 1818202714033618944 |
|---|---|
| author | Janyerson Dannys Pereira da Silva Gustavo Ballejo |
| author_facet | Janyerson Dannys Pereira da Silva Gustavo Ballejo |
| author_sort | Janyerson Dannys Pereira da Silva |
| collection | DOAJ |
| description | ABSTRACT Objective: To characterize the calcium influx pathways implicated in the sustained elevation of endothelial intracellular calcium concentration, required for the synthesis and release of relaxing factors. Methods: We evaluated the effect of the newly synthesized pyrazole derivatives, described as selective inhibitors for ORAI (BTP2/Pyr2 and Pyr6) and TRPC3 (Pyr3 and Pyr10) channels, upon endothelium- and extracellular calcium-dependent relaxations stimulated by acetylcholine and thapsigargin, in pre-constricted rat thoracic aortic rings. Results: Acetylcholine and thapsigargin responses were completely reverted by Pyr2 and Pyr6 (1 to 3μM). Pyr3 (0.3 to 3μM) caused a rapid reversal of acetylcholine (6.2±0.08mg.s−1) and thapsigargin (3.9±0.25mg.s−1) relaxations, whereas the more selective TRPC3 blocker Pyr10 (1 to 3μM) had no effect. The recently described TRPC4/5 selective blocker, ML204 (1 to 3μM), reverted completely acetylcholine relaxations, but minimally thapsigargin induced ones. Noteworthy, relaxations elicited by GSK1016790A (TRPV4 agonist) were unaffected by pyrazole compounds or ML204. After Pyr2 and Pyr6 pre-incubation, acetylcholine and thapsigargin evoked transient relaxations similar in magnitude and kinetics to those observed in the absence of extracellular calcium. Sodium nitroprusside relaxations as well as phenylephrine-induced contractions (denuded aorta) were not affected by any of pyrazole compounds (1 to 3μM). Conclusion: These observations revealed a previously unrecognized complexity in rat aorta endothelial calcium influx pathways, which result in production and release of nitric oxide. Pharmacologically distinguishable pathways mediate acetylcholine (ORAI/TRPC other than TRPC3/TRPC4 calcium-permeable channels) and thapsigargin (TRPC4 not required) induced calcium influx. |
| first_indexed | 2024-12-12T03:13:50Z |
| format | Article |
| id | doaj.art-f30dd572338f403ebce95b4ac0e94eec |
| institution | Directory Open Access Journal |
| issn | 2317-6385 |
| language | English |
| last_indexed | 2024-12-12T03:13:50Z |
| publisher | Instituto Israelita de Ensino e Pesquisa Albert Einstein |
| record_format | Article |
| series | Einstein (São Paulo) |
| spelling | doaj.art-f30dd572338f403ebce95b4ac0e94eec2022-12-22T00:40:21ZengInstituto Israelita de Ensino e Pesquisa Albert EinsteinEinstein (São Paulo)2317-638517310.31744/einstein_journal/2019ao4600S1679-45082019000300204Pharmacological characterization of the calcium influx pathways involved in nitric oxide production by endothelial cellsJanyerson Dannys Pereira da SilvaGustavo BallejoABSTRACT Objective: To characterize the calcium influx pathways implicated in the sustained elevation of endothelial intracellular calcium concentration, required for the synthesis and release of relaxing factors. Methods: We evaluated the effect of the newly synthesized pyrazole derivatives, described as selective inhibitors for ORAI (BTP2/Pyr2 and Pyr6) and TRPC3 (Pyr3 and Pyr10) channels, upon endothelium- and extracellular calcium-dependent relaxations stimulated by acetylcholine and thapsigargin, in pre-constricted rat thoracic aortic rings. Results: Acetylcholine and thapsigargin responses were completely reverted by Pyr2 and Pyr6 (1 to 3μM). Pyr3 (0.3 to 3μM) caused a rapid reversal of acetylcholine (6.2±0.08mg.s−1) and thapsigargin (3.9±0.25mg.s−1) relaxations, whereas the more selective TRPC3 blocker Pyr10 (1 to 3μM) had no effect. The recently described TRPC4/5 selective blocker, ML204 (1 to 3μM), reverted completely acetylcholine relaxations, but minimally thapsigargin induced ones. Noteworthy, relaxations elicited by GSK1016790A (TRPV4 agonist) were unaffected by pyrazole compounds or ML204. After Pyr2 and Pyr6 pre-incubation, acetylcholine and thapsigargin evoked transient relaxations similar in magnitude and kinetics to those observed in the absence of extracellular calcium. Sodium nitroprusside relaxations as well as phenylephrine-induced contractions (denuded aorta) were not affected by any of pyrazole compounds (1 to 3μM). Conclusion: These observations revealed a previously unrecognized complexity in rat aorta endothelial calcium influx pathways, which result in production and release of nitric oxide. Pharmacologically distinguishable pathways mediate acetylcholine (ORAI/TRPC other than TRPC3/TRPC4 calcium-permeable channels) and thapsigargin (TRPC4 not required) induced calcium influx.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1679-45082019000300204&lng=en&tlng=enCélulas endoteliaisCálcio/metabolismoProteína ORAI1Canais de cátion TRPCAortaÓxido nítricoRatosRelaxamento |
| spellingShingle | Janyerson Dannys Pereira da Silva Gustavo Ballejo Pharmacological characterization of the calcium influx pathways involved in nitric oxide production by endothelial cells Einstein (São Paulo) Células endoteliais Cálcio/metabolismo Proteína ORAI1 Canais de cátion TRPC Aorta Óxido nítrico Ratos Relaxamento |
| title | Pharmacological characterization of the calcium influx pathways involved in nitric oxide production by endothelial cells |
| title_full | Pharmacological characterization of the calcium influx pathways involved in nitric oxide production by endothelial cells |
| title_fullStr | Pharmacological characterization of the calcium influx pathways involved in nitric oxide production by endothelial cells |
| title_full_unstemmed | Pharmacological characterization of the calcium influx pathways involved in nitric oxide production by endothelial cells |
| title_short | Pharmacological characterization of the calcium influx pathways involved in nitric oxide production by endothelial cells |
| title_sort | pharmacological characterization of the calcium influx pathways involved in nitric oxide production by endothelial cells |
| topic | Células endoteliais Cálcio/metabolismo Proteína ORAI1 Canais de cátion TRPC Aorta Óxido nítrico Ratos Relaxamento |
| url | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1679-45082019000300204&lng=en&tlng=en |
| work_keys_str_mv | AT janyersondannyspereiradasilva pharmacologicalcharacterizationofthecalciuminfluxpathwaysinvolvedinnitricoxideproductionbyendothelialcells AT gustavoballejo pharmacologicalcharacterizationofthecalciuminfluxpathwaysinvolvedinnitricoxideproductionbyendothelialcells |