Comparison of MEK/ERK pathway inhibitors on the upregulation of vascular G-protein coupled receptors in rat cerebral arteries
(2010) In European Journal of Pharmacology 644(1-3). p.128-137- Abstract
- Organ culture is an in vitro method for investigating cellular mechanisms involved in upregulation of vasocontractile G-protein coupled receptors. We hypothesize that mitogen-activated-protein kinase (MEK) and/or extracellular-signal-regulated kinase (ERK) specific inhibitors will attenuate the G-protein coupled receptor expression following organ culture. Rat cerebral arteries were incubated 48 h in the presence of MEK/ERK specific inhibitors U0126, PD98059, SL327, or AG126 for different time periods. Contractile responses by activation of endothelin receptor type A and type B. serotonin receptor 5-FIT18, prostanoid TP receptor, and angiotensin II receptor type 1 and type 2 were investigated. Results were verified by measurement of mRNA... (More)
- Organ culture is an in vitro method for investigating cellular mechanisms involved in upregulation of vasocontractile G-protein coupled receptors. We hypothesize that mitogen-activated-protein kinase (MEK) and/or extracellular-signal-regulated kinase (ERK) specific inhibitors will attenuate the G-protein coupled receptor expression following organ culture. Rat cerebral arteries were incubated 48 h in the presence of MEK/ERK specific inhibitors U0126, PD98059, SL327, or AG126 for different time periods. Contractile responses by activation of endothelin receptor type A and type B. serotonin receptor 5-FIT18, prostanoid TP receptor, and angiotensin II receptor type 1 and type 2 were investigated. Results were verified by measurement of mRNA with real time PCR and by protein immunohistochemistry. Organ culture induced transcriptional upregulation of endothelin ETB receptor and of serotonin 5-HTIB receptor on translational level and increased respective contractions. The prostanoid TP receptor mediated contraction curve was left-wards shifted by organ culture. Organ culture was associated with elevated pERK1/2 in the vascular smooth muscle cells: the MEK1/2 inhibitor U0126 attenuated the endothelin ETB receptor mediated contraction at post-translational level or by changing the receptor affinities. The serotonin 5-HT,B receptor and prostanoid TP receptor mediated contractions were abolished by U0126. Administration of U0126 6 h after start of incubation blocked the receptor upregulation. In conclusion, MEK specific inhibitor U0126 is a potent inhibitor of G-protein coupled receptor alteration seen during organ culture. Given the ability to inhibit G-protein coupled receptor alteration at the clinically relevant time-point 6 h post incubation makes it an attractive therapeutic agent for in vivo studies. (C) 2010 Elsevier By. All rights reserved. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/1727248
- author
- Sandhu, Hardip ; Ansar, Saema and Edvinsson, Lars LU
- organization
- publishing date
- 2010
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- MEK/ERK pathway, U0126, Cerebral arteries, Organ culture, G-protein coupled receptor upregulation, Vascular smooth muscle cells
- in
- European Journal of Pharmacology
- volume
- 644
- issue
- 1-3
- pages
- 128 - 137
- publisher
- Elsevier
- external identifiers
-
- wos:000281935800020
- scopus:77955767817
- pmid:20615400
- ISSN
- 1879-0712
- DOI
- 10.1016/j.ejphar.2010.06.053
- language
- English
- LU publication?
- yes
- id
- c8517ef7-5b04-443f-8d1a-de32ab50fd58 (old id 1727248)
- date added to LUP
- 2016-04-01 10:42:38
- date last changed
- 2024-01-06 23:17:54
@article{c8517ef7-5b04-443f-8d1a-de32ab50fd58,
abstract = {{Organ culture is an in vitro method for investigating cellular mechanisms involved in upregulation of vasocontractile G-protein coupled receptors. We hypothesize that mitogen-activated-protein kinase (MEK) and/or extracellular-signal-regulated kinase (ERK) specific inhibitors will attenuate the G-protein coupled receptor expression following organ culture. Rat cerebral arteries were incubated 48 h in the presence of MEK/ERK specific inhibitors U0126, PD98059, SL327, or AG126 for different time periods. Contractile responses by activation of endothelin receptor type A and type B. serotonin receptor 5-FIT18, prostanoid TP receptor, and angiotensin II receptor type 1 and type 2 were investigated. Results were verified by measurement of mRNA with real time PCR and by protein immunohistochemistry. Organ culture induced transcriptional upregulation of endothelin ETB receptor and of serotonin 5-HTIB receptor on translational level and increased respective contractions. The prostanoid TP receptor mediated contraction curve was left-wards shifted by organ culture. Organ culture was associated with elevated pERK1/2 in the vascular smooth muscle cells: the MEK1/2 inhibitor U0126 attenuated the endothelin ETB receptor mediated contraction at post-translational level or by changing the receptor affinities. The serotonin 5-HT,B receptor and prostanoid TP receptor mediated contractions were abolished by U0126. Administration of U0126 6 h after start of incubation blocked the receptor upregulation. In conclusion, MEK specific inhibitor U0126 is a potent inhibitor of G-protein coupled receptor alteration seen during organ culture. Given the ability to inhibit G-protein coupled receptor alteration at the clinically relevant time-point 6 h post incubation makes it an attractive therapeutic agent for in vivo studies. (C) 2010 Elsevier By. All rights reserved.}},
author = {{Sandhu, Hardip and Ansar, Saema and Edvinsson, Lars}},
issn = {{1879-0712}},
keywords = {{MEK/ERK pathway; U0126; Cerebral arteries; Organ culture; G-protein coupled receptor upregulation; Vascular smooth muscle cells}},
language = {{eng}},
number = {{1-3}},
pages = {{128--137}},
publisher = {{Elsevier}},
series = {{European Journal of Pharmacology}},
title = {{Comparison of MEK/ERK pathway inhibitors on the upregulation of vascular G-protein coupled receptors in rat cerebral arteries}},
url = {{http://dx.doi.org/10.1016/j.ejphar.2010.06.053}},
doi = {{10.1016/j.ejphar.2010.06.053}},
volume = {{644}},
year = {{2010}},
}