Endothelin-1 reduces cAMP generation by iloprost from cerebral microvascular endothelial cells

M. A. Yakubu, Charles Leffler

Research output: Contribution to journalArticle

Abstract

We have investigated the effects of pretreatment of cerebral microvascular endothelial cell culture with endothelin-1 (ET-1) on iloprost-induced generation of cyclic 3,5-adenosine monophosphate (cAMP). We tested the hypothesis that ET-1 inhibits cAMP production and that pretreatment of cultured endothelial cells with ET-1 will attenuate the iloprost-induced production of cAMP. Confluent cells were treated with PBS, ET-1, or iloprost alone. To assess effects of ET-1 pretreatment on cAMP production, cultured endothelial cells were incubated with iloprost following 30 min pretreatment with ET-1 and samples collected for the determination of cAMP. At the end of the incubation the media was collected and the cells were scraped following treatment with 0.1N HCl. The samples were analyzed for cAMP content using RIA. Incubation of endothelial cells with iloprost 10-8 and 10-6 M, increased levels of cAMP production by the cells from 14±3 in the control to 37±6 and 128±32 pmol/mg protein respectively. While ET-1 10-8 and 10-6 M resulted in the accumulation of 7±3 and 12±5 pmol/mg protein cAMP. Pretreatment with ET-1 (10-8 M) attenuated iloprost-induced increase in cAMP compared to iloprost alone (23±5 versus 37±6 and 54±8 versus 128±32 pmol/mg protein). Similarly, prior exposure of pial arterioles to ET-1 (10-9 M) attenuated iloprost-induced dilation. This study shows that prior exposure of endothelial cells or pial arterioles to ET-1 attenuates responses to iloprost. The mechanism(s) behind the altered cerebral microcirculation following conditions with increased ET-1 may involve reduced sensitivity of pial arterioles to the regulation of cAMP by iloprost.

Original languageEnglish (US)
JournalFASEB Journal
Volume12
Issue number5
StatePublished - Mar 20 1998

Fingerprint

Iloprost
endothelins
Endothelial cells
cyclic AMP
Endothelin-1
Adenosine Monophosphate
endothelial cells
Endothelial Cells
pretreatment
Arterioles
cultured cells
Cultured Cells
Cyclic AMP
Microcirculation
Proteins
proteins
cells
Cell culture
Dilatation
cell culture

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Genetics

Cite this

Endothelin-1 reduces cAMP generation by iloprost from cerebral microvascular endothelial cells. / Yakubu, M. A.; Leffler, Charles.

In: FASEB Journal, Vol. 12, No. 5, 20.03.1998.

Research output: Contribution to journalArticle

@article{a99d1036705a4fdeb827808ec578ac01,
title = "Endothelin-1 reduces cAMP generation by iloprost from cerebral microvascular endothelial cells",
abstract = "We have investigated the effects of pretreatment of cerebral microvascular endothelial cell culture with endothelin-1 (ET-1) on iloprost-induced generation of cyclic 3,5-adenosine monophosphate (cAMP). We tested the hypothesis that ET-1 inhibits cAMP production and that pretreatment of cultured endothelial cells with ET-1 will attenuate the iloprost-induced production of cAMP. Confluent cells were treated with PBS, ET-1, or iloprost alone. To assess effects of ET-1 pretreatment on cAMP production, cultured endothelial cells were incubated with iloprost following 30 min pretreatment with ET-1 and samples collected for the determination of cAMP. At the end of the incubation the media was collected and the cells were scraped following treatment with 0.1N HCl. The samples were analyzed for cAMP content using RIA. Incubation of endothelial cells with iloprost 10-8 and 10-6 M, increased levels of cAMP production by the cells from 14±3 in the control to 37±6 and 128±32 pmol/mg protein respectively. While ET-1 10-8 and 10-6 M resulted in the accumulation of 7±3 and 12±5 pmol/mg protein cAMP. Pretreatment with ET-1 (10-8 M) attenuated iloprost-induced increase in cAMP compared to iloprost alone (23±5 versus 37±6 and 54±8 versus 128±32 pmol/mg protein). Similarly, prior exposure of pial arterioles to ET-1 (10-9 M) attenuated iloprost-induced dilation. This study shows that prior exposure of endothelial cells or pial arterioles to ET-1 attenuates responses to iloprost. The mechanism(s) behind the altered cerebral microcirculation following conditions with increased ET-1 may involve reduced sensitivity of pial arterioles to the regulation of cAMP by iloprost.",
author = "Yakubu, {M. A.} and Charles Leffler",
year = "1998",
month = "3",
day = "20",
language = "English (US)",
volume = "12",
journal = "FASEB Journal",
issn = "0892-6638",
publisher = "FASEB",
number = "5",

}

TY - JOUR

T1 - Endothelin-1 reduces cAMP generation by iloprost from cerebral microvascular endothelial cells

AU - Yakubu, M. A.

AU - Leffler, Charles

PY - 1998/3/20

Y1 - 1998/3/20

N2 - We have investigated the effects of pretreatment of cerebral microvascular endothelial cell culture with endothelin-1 (ET-1) on iloprost-induced generation of cyclic 3,5-adenosine monophosphate (cAMP). We tested the hypothesis that ET-1 inhibits cAMP production and that pretreatment of cultured endothelial cells with ET-1 will attenuate the iloprost-induced production of cAMP. Confluent cells were treated with PBS, ET-1, or iloprost alone. To assess effects of ET-1 pretreatment on cAMP production, cultured endothelial cells were incubated with iloprost following 30 min pretreatment with ET-1 and samples collected for the determination of cAMP. At the end of the incubation the media was collected and the cells were scraped following treatment with 0.1N HCl. The samples were analyzed for cAMP content using RIA. Incubation of endothelial cells with iloprost 10-8 and 10-6 M, increased levels of cAMP production by the cells from 14±3 in the control to 37±6 and 128±32 pmol/mg protein respectively. While ET-1 10-8 and 10-6 M resulted in the accumulation of 7±3 and 12±5 pmol/mg protein cAMP. Pretreatment with ET-1 (10-8 M) attenuated iloprost-induced increase in cAMP compared to iloprost alone (23±5 versus 37±6 and 54±8 versus 128±32 pmol/mg protein). Similarly, prior exposure of pial arterioles to ET-1 (10-9 M) attenuated iloprost-induced dilation. This study shows that prior exposure of endothelial cells or pial arterioles to ET-1 attenuates responses to iloprost. The mechanism(s) behind the altered cerebral microcirculation following conditions with increased ET-1 may involve reduced sensitivity of pial arterioles to the regulation of cAMP by iloprost.

AB - We have investigated the effects of pretreatment of cerebral microvascular endothelial cell culture with endothelin-1 (ET-1) on iloprost-induced generation of cyclic 3,5-adenosine monophosphate (cAMP). We tested the hypothesis that ET-1 inhibits cAMP production and that pretreatment of cultured endothelial cells with ET-1 will attenuate the iloprost-induced production of cAMP. Confluent cells were treated with PBS, ET-1, or iloprost alone. To assess effects of ET-1 pretreatment on cAMP production, cultured endothelial cells were incubated with iloprost following 30 min pretreatment with ET-1 and samples collected for the determination of cAMP. At the end of the incubation the media was collected and the cells were scraped following treatment with 0.1N HCl. The samples were analyzed for cAMP content using RIA. Incubation of endothelial cells with iloprost 10-8 and 10-6 M, increased levels of cAMP production by the cells from 14±3 in the control to 37±6 and 128±32 pmol/mg protein respectively. While ET-1 10-8 and 10-6 M resulted in the accumulation of 7±3 and 12±5 pmol/mg protein cAMP. Pretreatment with ET-1 (10-8 M) attenuated iloprost-induced increase in cAMP compared to iloprost alone (23±5 versus 37±6 and 54±8 versus 128±32 pmol/mg protein). Similarly, prior exposure of pial arterioles to ET-1 (10-9 M) attenuated iloprost-induced dilation. This study shows that prior exposure of endothelial cells or pial arterioles to ET-1 attenuates responses to iloprost. The mechanism(s) behind the altered cerebral microcirculation following conditions with increased ET-1 may involve reduced sensitivity of pial arterioles to the regulation of cAMP by iloprost.

UR - http://www.scopus.com/inward/record.url?scp=33749224069&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33749224069&partnerID=8YFLogxK

M3 - Article

VL - 12

JO - FASEB Journal

JF - FASEB Journal

SN - 0892-6638

IS - 5

ER -