Abstract
1. Nitric oxide (NO) regulates cyclo-oxygenase (COX) activity in various cell systems and reports conflict in regard to its stimulatory versus inhibitory role. Incubation of human umbilical vein endothelial cells (HUVEC) with SIN-1 (3-morpholinosydnonimine), a donor of NO, resulted in a rapid and dose-dependent increase in the expression of COX-2 as analysed by Western and Northern blotting. 2. Incubation of HUVEC with SIN-1 and interleukine (IL)-1α resulted in increased induction of COX-2 compared with IL-1α alone and corresponded to an additive effect. The COX-2 induction was dependent on a de novo synthesis since cycloheximide, an inhibitor of protein synthesis, blocked the enzyme expression. The increase in COX-2 expression was not accompanied by a corresponding change in prostaglandin (PG) production. However, the COX activity was partially recovered when immunoprecipitated COX-2 was incubated with arachidonic acid and haematin. 3. Peroxynitrite, a highly reactive nitrogen molecule derived from the interaction of NO and superoxide anion, significantly increased COX-2 expression. Under these conditions and within the limit of detection of the antibody, selective antibody for nitrotyrosine failed to detect nitrated COX-2 in immunoprecipitated COX-2 when cells where incubated with SIN-1 or SIN-1 + IL-1α. 4. Ro 31-8220, a specific inhibitor of protein kinase (PK) C, blocked the induction of COX-2. Also, SB203580, the selective inhibitor of p38 MAP kinase, strongly blocked the induction of COX-2 by SIN-1 in the presence or absence of IL-1α, whereas the MEK-1 inhibitor, PD 98059, affected it to a lesser extent. These data demonstrate that SIN-1 induces COX-2 in HUVEC in the absence of PG formation and suggest a complex regulation of COX-2 expression and PG formation by NO in endothelial cells.
| Original language | English |
|---|---|
| Pages (from-to) | 1163-1171 |
| Number of pages | 9 |
| Journal | British Journal of Pharmacology |
| Volume | 133 |
| Issue number | 7 |
| Publication status | Published - 2001 |
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Keywords
- COX-2
- Endothelial cells
- MAP kinases
- Nitric oxide (NO)
- Prostacyclin
- SIN-1
ASJC Scopus subject areas
- Pharmacology
Cite this
Induction of cyclo-oxygenase-2 in human endothelial cells by SIN-1 in the absence of prostaglandin production. / Eligini, Sonia; Habib, Aïda; Lebret, Marilyne; Créminon, Christophe; Lévy-Toledano, Sylviane; Maclouf, Jacques.
In: British Journal of Pharmacology, Vol. 133, No. 7, 2001, p. 1163-1171.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Induction of cyclo-oxygenase-2 in human endothelial cells by SIN-1 in the absence of prostaglandin production
AU - Eligini, Sonia
AU - Habib, Aïda
AU - Lebret, Marilyne
AU - Créminon, Christophe
AU - Lévy-Toledano, Sylviane
AU - Maclouf, Jacques
PY - 2001
Y1 - 2001
N2 - 1. Nitric oxide (NO) regulates cyclo-oxygenase (COX) activity in various cell systems and reports conflict in regard to its stimulatory versus inhibitory role. Incubation of human umbilical vein endothelial cells (HUVEC) with SIN-1 (3-morpholinosydnonimine), a donor of NO, resulted in a rapid and dose-dependent increase in the expression of COX-2 as analysed by Western and Northern blotting. 2. Incubation of HUVEC with SIN-1 and interleukine (IL)-1α resulted in increased induction of COX-2 compared with IL-1α alone and corresponded to an additive effect. The COX-2 induction was dependent on a de novo synthesis since cycloheximide, an inhibitor of protein synthesis, blocked the enzyme expression. The increase in COX-2 expression was not accompanied by a corresponding change in prostaglandin (PG) production. However, the COX activity was partially recovered when immunoprecipitated COX-2 was incubated with arachidonic acid and haematin. 3. Peroxynitrite, a highly reactive nitrogen molecule derived from the interaction of NO and superoxide anion, significantly increased COX-2 expression. Under these conditions and within the limit of detection of the antibody, selective antibody for nitrotyrosine failed to detect nitrated COX-2 in immunoprecipitated COX-2 when cells where incubated with SIN-1 or SIN-1 + IL-1α. 4. Ro 31-8220, a specific inhibitor of protein kinase (PK) C, blocked the induction of COX-2. Also, SB203580, the selective inhibitor of p38 MAP kinase, strongly blocked the induction of COX-2 by SIN-1 in the presence or absence of IL-1α, whereas the MEK-1 inhibitor, PD 98059, affected it to a lesser extent. These data demonstrate that SIN-1 induces COX-2 in HUVEC in the absence of PG formation and suggest a complex regulation of COX-2 expression and PG formation by NO in endothelial cells.
AB - 1. Nitric oxide (NO) regulates cyclo-oxygenase (COX) activity in various cell systems and reports conflict in regard to its stimulatory versus inhibitory role. Incubation of human umbilical vein endothelial cells (HUVEC) with SIN-1 (3-morpholinosydnonimine), a donor of NO, resulted in a rapid and dose-dependent increase in the expression of COX-2 as analysed by Western and Northern blotting. 2. Incubation of HUVEC with SIN-1 and interleukine (IL)-1α resulted in increased induction of COX-2 compared with IL-1α alone and corresponded to an additive effect. The COX-2 induction was dependent on a de novo synthesis since cycloheximide, an inhibitor of protein synthesis, blocked the enzyme expression. The increase in COX-2 expression was not accompanied by a corresponding change in prostaglandin (PG) production. However, the COX activity was partially recovered when immunoprecipitated COX-2 was incubated with arachidonic acid and haematin. 3. Peroxynitrite, a highly reactive nitrogen molecule derived from the interaction of NO and superoxide anion, significantly increased COX-2 expression. Under these conditions and within the limit of detection of the antibody, selective antibody for nitrotyrosine failed to detect nitrated COX-2 in immunoprecipitated COX-2 when cells where incubated with SIN-1 or SIN-1 + IL-1α. 4. Ro 31-8220, a specific inhibitor of protein kinase (PK) C, blocked the induction of COX-2. Also, SB203580, the selective inhibitor of p38 MAP kinase, strongly blocked the induction of COX-2 by SIN-1 in the presence or absence of IL-1α, whereas the MEK-1 inhibitor, PD 98059, affected it to a lesser extent. These data demonstrate that SIN-1 induces COX-2 in HUVEC in the absence of PG formation and suggest a complex regulation of COX-2 expression and PG formation by NO in endothelial cells.
KW - COX-2
KW - Endothelial cells
KW - MAP kinases
KW - Nitric oxide (NO)
KW - Prostacyclin
KW - SIN-1
UR - http://www.scopus.com/inward/record.url?scp=0034887817&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0034887817&partnerID=8YFLogxK
M3 - Article
C2 - 11487528
AN - SCOPUS:0034887817
VL - 133
SP - 1163
EP - 1171
JO - British Journal of Pharmacology
JF - British Journal of Pharmacology
SN - 0007-1188
IS - 7
ER -