This Article Abstract Full Text (PDF) All Versions of this Article: 45/1/126    most recent 01.HYP.0000150159.48992.11v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Kohlstedt, K. Articles by Fleming, I. Search for Related Content PubMed PubMed Citation Articles by Kohlstedt, K. Articles by Fleming, I. Pubmed/NCBI databases Gene GEO Profiles HomoloGene UniGene Compound via MeSH Substance via MeSH Hazardous Substances DB CELECOXIB Related Collections Cardiovascular Pharmacology Other Vascular biology ACE/Angiotension receptors Cell signalling/signal transduction Gene expression

(Hypertension. 2005;45:126.)
© 2005 American Heart Association, Inc.

Scientific Contributions

Signaling via the Angiotensin-Converting Enzyme Enhances the Expression of Cyclooxygenase-2 in Endothelial Cells

Karin Kohlstedt; Rudi Busse; Ingrid Fleming

From the Vascular Signaling Group, Institut für Kardiovaskuläre Physiologie, Johann Wolfgang Goethe-Universität, Frankfurt am Main, Germany.

Correspondence to Ingrid Fleming, PhD, Vascular Signaling Group, Institut für Kardiovaskuläre Physiologie, Johann Wolfgang Goethe-Universität, Theodor-Stern-Kai 7, D-60590 Frankfurt am Main, Germany. E-mail fleming{at}em.uni-frankfurt.de

	Abstract

Top Abstract Introduction Materials and Methods Results Discussion References

 
Angiotensin-converting enzyme (ACE) inhibitors elicit outside-in signaling via ACE in endothelial cells. This involves the CK2-mediated phosphorylation of ACE on Ser¹²⁷⁰ and the activation of the c-Jun N-terminal kinase (JNK)/c-Jun pathway, resulting in an enhanced endothelial ACE expression. Because cyclooxygenase-2 (COX-2) expression is reported to be increased in subjects treated with ACE inhibitors, we determined the role of ACE signaling in this phenomenon and the transcription factors involved. In lungs from mice treated with the ACE inhibitor ramipril for 5 days, COX-2 expression was increased. A similar (1.5- to 2-fold) increase in COX-2 protein was detected in primary cultures of human endothelial cells treated with ramiprilat. In an endothelial cell line stably expressing human somatic ACE, ramiprilat increased COX-2 promoter activity, an effect not observed in ACE-deficient cells or cells expressing a nonphosphorylatable ACE mutant (S1270A). The ramiprilat-induced, ACE-dependent increase in COX-2 expression and promoter activity (both 1.5- to 2-fold greater than control) was prevented by the inhibition of JNK. Ramiprilat significantly enhanced the DNA binding activity of activator protein-1 in cells expressing ACE but not S1270A ACE. Activator protein-1 decoy oligonucleotides prevented the ACE inhibitor-induced increase in COX-2 promoter activity and protein expression. As a consequence of the ramiprilat-induced increase in COX-2 expression, prostacyclin and prostaglandin E₂, but not thromboxane A₂, production was increased and was inhibited by the COX-2 inhibitor celecoxib. These results indicate that ACE signaling may underlie the increase in COX-2 and prostacyclin levels in patients treated with ACE inhibitors.

Key Words: angiotensin-converting enzyme • cyclooxygenase • endothelium • prostacyclin • ramipril • signal transduction

	Introduction

Top Abstract Introduction Materials and Methods Results Discussion References

 
Inhibitors of the angiotensin-converting enzyme (ACE), such as ramipril, exert beneficial effects on endothelial function and vascular remodeling^1,2 and protect against the progression of atherosclerosis and the occurrence of cardiovascular events in humans.³ Although the latter effects are generally attributed to a decrease in the ACE-mediated generation of angiotensin II and the accumulation of bradykinin, a number of the effects of ACE inhibitors cannot be accounted for by inhibition of the enzyme per se.^4–6

One consequence of prolonged ACE inhibitor therapy is an increase in the expression of the enzyme itself. Increased ACE levels have been demonstrated in lung tissue and plasma from ACE inhibitor-treated rats^7,8 and in the serum from patients who distinctly benefit from ACE inhibitor therapy.^9,10 The finding that ACE inhibitors enhance the phosphorylation of ACE on Ser¹²⁷⁰ within the short cytoplasmic tail of the enzyme and thus induce the phosphorylation and nuclear translocation of c-Jun,¹¹ together with the report that the increase in ACE expression detected in phorbol ester-stimulated endothelial cells can be attributed to the activity of an activator protein-1 (AP-1) complex containing a c-Jun homodimer,¹² suggests that a novel ACE-dependent signaling cascade can affect gene expression.

The expression of cyclooxygenase-2 (COX-2) is regulated by c-Jun,¹³ and because prostacyclin (PGI₂) production is increased in subjects treated with ACE inhibitors,^14,15 we hypothesized that an ACE signaling pathway may affect COX-2 expression and thus vasodilator prostanoid production. To assess the role of ACE signaling in the regulation of COX-2 expression, we determined the effects of ramipril/ramiprilat in vivo (mouse lung), in primary cultures of human endothelial cells that express ACE, as well as in 3 porcine aortic endothelial cell lines. The latter endothelial cells were either ACE-deficient or overexpressed either human somatic ACE or a nonphosphorylatable ACE mutant in which Ser¹²⁷⁰ was mutated to alanine (S1270A).

	Materials and Methods

Top Abstract Introduction Materials and Methods Results Discussion References

 
Celecoxib was a generous gift from Professor G. Geisslinger (Pharmazentrum, Frankfurt), and the JNK inhibitor SP600125 was from Tocris (Bristol, UK). All other substances were obtained from Sigma.

Cell Culture
Human umbilical vein endothelial cells were isolated and cultured as described.¹⁶ Because ACE expression decreases with time in culture, all of the experiments in human endothelial cells were performed using primary cultures. Porcine aortic endothelial cells stably transfected with ACE or the S1270A ACE mutant were generated and cultured as described.¹⁷ Although the porcine endothelial cells no longer endogenously expressed ACE or functional angiotensin II and bradykinin receptors, they expressed a number of characteristic endothelial cell proteins (von Willebrand factor, CD31, the endothelial nitric oxide synthase and vascular endothelial cadherin).

Animals
To analyze the effect of ACE inhibition on COX-2 expression in vivo in male mice (C57 black 6; 6 weeks; Charles River, Wilmington, Mass), ramipril (5 mg/kg per day) was included in the drinking water over 5 days. Thereafter, the mice were anesthetized (isofuran 1.5%) and euthanized by a transverse cut through the large abdominal vessel. The lungs were perfused rapidly with ice-cold phosphate-buffered saline and snap-frozen homogenized as described.¹¹

Immunoblotting
Cells or lung homogenates were either boiled in SDS-sample buffer or lysed in nonidet lysis buffer, left on ice for 10 minutes, and centrifuged at 10 000g for 10 minutes. Supernatants were heated in SDS-PAGE sample buffer and separated by SDS-PAGE.¹⁷ Proteins were detected using their respective antibodies and were visualized by enhanced chemiluminescence using a commercially available kit (Amersham). The COX-2 and CD31 antibodies used for Western blotting were from Santa Cruz Biotechnology, the tubulin antibody from Dianova (Hamburg, Germany), and the ACE monoclonal antibody was provided by Dr Peter Bünning (Aventis, Frankfurt, Germany).

Luciferase Reporter Gene Assay
Cells were transiently transfected with a COX-2 promoter construct (kindly provided by Margarete Goppelt-Struebe, Erlangen, Germany) together with a LacZ construct (Invitrogen, Karlsruhe, Germany) using calcium phosphate precipitation as described.¹⁷ After stimulation, the cells were lysed and luciferase activity was assayed using a commercially available kit (Promega, Mannheim, Germany). The values obtained were normalized to ß-galactosidase activity (Tropix, Bedford, Mass) or to protein content. In some experiments, cells were cotransfected with flag-tagged dominant-negative JNK (pcDNA3.1Flag-JNK1-APF), wild-type JNK (pcDNA3.1Flag-JNK1), or empty vector (pcDNA3.1)¹⁸ 24 hours before transfection with the promoter construct.

Prostaglandin Assays
6-Keto-PG F₁ (6-Keto-PGF₁) and prostaglandin E₂ (PGE₂) levels were determined in the supernatants of primary cultures of human endothelial cells using commercially available kits (PGI₂: Amersham Bioscience; PGE₂: Assay Designs Inc).

Electrophoretic Mobility Shift Assay
Double-stranded oligonucleotides containing the sequence of the binding site for AP-1 (5'-CGCTTGATGAGTCAGCCGGAA-3', Promega, Wis) were labeled with [³²P]-ATP using a Ready-to-go DNA-labeling Kit (Amersham Biosciences, Piscataway, NJ). Nuclear proteins (6 to 10 µg) were incubated with the ³²P-labeled oligonucleotides (10 000 counts/sample) and protein–DNA complexes were resolved by electrophoresis as described.¹⁹ Densitometric analysis of the autoradiographs was performed after nonsaturating exposures and the values expressed as the percentage of the binding activity in unstimulated cell extracts.

AP-1 Decoy Oligonucleotides
To assess the role of AP-1 in the ramiprilat-induced increase in COX-2 expression, endothelial cells were incubated with AP-1 decoy oligonucleotides as described.¹⁹ The AP-1 decoy oligonucleotides (5'-CGCTTGATGAGTCAGCCGGAATTTTTTCCGGCTGACTCATCAAGCG-3') and the AP-1 scrambled oligonucleotides (5'-CGCTTGATTACTTAGCCGGAATTTTTTCCGGCTAAGTAATCAAGCG-3') were synthesized by BioSpring (Frankfurt, Germany). Preliminary experiments assessing the effects of the oligonucleotides on the DNA-binding activity of AP-1 and NF-B were performed to determine the specificity of the oligonucleotides at the concentration used.

Statistical Analysis
Data are expressed as mean±SEM and statistical evaluation was performed using Student t test for unpaired data or 1-way ANOVA followed by a Bonferroni t test when appropriate. Values of P<0.05 were considered statistically significant.

	Results

Top Abstract Introduction Materials and Methods Results Discussion References

 
Effect of an ACE Inhibitor on the Expression of COX-2 in Primary Cultures of Human Endothelial Cells and in the Mouse Lung
To assess the effect of ramiprilat on the expression of COX-2 in endothelial cells, primary cultures of human umbilical vein endothelial cells were stimulated with ramiprilat (100 nmol/L, 4 to 72 hours) and harvested in SDS sample buffer. Ramiprilat induced a significant increase in the expression of COX-2, which was first evident 4 hours after the addition of the ACE inhibitor and remained elevated over the rest of the experimental period (72 hours; Figure 1A). Although COX-1 was expressed in human endothelial cells, ramiprilat failed to increase COX-1 expression (data not shown). A significant increase in the expression of COX-2 was also detected in the lungs of mice treated with ramipril over 5 days (Figure 1B).

View larger version (47K): [in this window] [in a new window]   Figure 1. Effect of the ACE inhibitor ramiprilat on the expression of COX-2 in vitro and in vivo. A, Time course of the ramiprilat (100 nmol/L)-induced increase in COX-2 expression in primary cultures of human umbilical vein endothelial cells. B, Effect of prolonged ramipril treatment (Rami, 5 mg/kg per day) for 5 days on the expression of COX-2 in mouse lungs. COX-2 levels were normalized to either CD31 or tubulin. The bar graphs summarize data obtained in 6 to 8 independent experiments. *P<0.05, **P<0.01 vs control (CTL).

Effect of Ramiprilat on COX-2 Promoter Activity in Porcine Aortic Endothelial Cells
To determine whether the effect of ramiprilat on COX-2 protein expression in vivo and in vitro was mediated by the ACE signaling pathway, we compared the effect of ramiprilat on COX-2 promoter activity in porcine aortic endothelial cells deficient in ACE and in cells stably expressing either human somatic ACE or the nonphosphorylatable ACE mutant (S1270A). Promoter activity was assessed as the cell lines did not express COX-2 under basal conditions or after cell stimulation (data not shown).

Ramiprilat (100 nmol/L, 24 hours) significantly enhanced COX-2 promoter activity in cells expressing human somatic ACE (Figure 2A). However, the ACE inhibitor failed to affect COX-2 promoter activity in ACE-deficient cells or in cells expressing the S1270A ACE mutant. The effect of ramiprilat on COX-2 promoter activity was comparable with that of interleukin (IL)-1ß, but weaker than that of phorbol 12-myristate 13-acetate (Figure 2B).

View larger version (14K): [in this window] [in a new window]   Figure 2. Effect of ramiprilat on the activity of the COX-2 promoter. A, COX-2 promoter activity was assessed in porcine endothelial cell lines overexpressing either human somatic ACE (ACE) or the S1270A ACE mutant (S1270A), or in ACE-deficient endothelial cells (–ACE). Experiments were performed in the absence (C) and presence of ramiprilat (R, 100 nmol/L, 24 hours). B, COX-2 promoter activity was assessed in ACE overexpressing porcine endothelial cells in the absence (CTL) and presence of either ramiprilat (Rami, 100 nmol/L, 24 hours), IL-1ß (20 ng/mL, 24 hours), or PMA (20 ng/mL, 24 hours). COX-2 promoter activity was quantified as luciferase activity relative to the activity measured in control cells. The results shown are the mean±SEM of data obtained in 8 to 10 separate experiments, *P<0.05, **P<0.01 vs control (C, CTL).

Effect of JNK Inhibition on COX-2 Upregulation by Ramiprilat in Endothelial Cells
Pretreatment of primary cultures of human endothelial cells with the JNK inhibitor, SP600125 (5 µmol/L), prevented the ramiprilat-induced (100 nmol/L, 24 hours) increase in COX-2 expression (Figure 3A).

View larger version (37K): [in this window] [in a new window]   Figure 3. Effect of JNK inhibition and dominant-negative JNK on the ramiprilat-induced increase in COX-2 promoter activity and expression. A, Western blots showing the time-dependent effect of ramiprilat (Rami, 100 nmol/L) on the expression of COX-2 in human endothelial cells in the absence and presence of the JNK inhibitor, SP600125 (5 µmol/L). COX-2 protein levels were normalized to ß-actin levels and the bar graph summarizes data obtained in 5 independent experiments. B, Representative Western blot showing the expression of the endogenous and flag-tagged JNKs (Flag-JNK) in cells transfected with an empty plasmid (vector), dominant-negative JNK (dnJNK), or wild-type JNK (wtJNK). The bar graph summarizes the effect of ramiprilat (R, 100 nmol/L, 24 hours) on COX-2 promoter activity in ACE-overexpressing porcine endothelial cells transfected with the appropriate plasmid. Promoter activity was normalized relative to the levels measured in control (C, CTL), solvent-treated cells; *P<0.05 vs the respective CTL.

To determine whether the ramiprilat-induced increase in JNK activity can enhance the activity of the COX-2 promoter, experiments were performed in the ACE-expressing endothelial cell line. Transfection of these cells with a control vector or wild-type JNK failed to affect the ramiprilat-induced increase in promoter activity, whereas transfection with a dominant-negative JNK completely prevented the effect of the ACE inhibitor (Figure 3B).

Role of AP-1 in the Ramiprilat-Induced Upregulation of COX-2
Because COX-2 expression in endothelial cells is reportedly influenced by the transcription factors AP-1 and the cAMP-response element binding protein,²⁰ we determined the effect of ramiprilat on the activity of these transcription factors in the ACE-overexpressing endothelial cell line. Ramiprilat did not affect the DNA binding activity of cAMP-response element binding protein (data not shown), but significantly enhanced the DNA binding activity of AP-1 (Figure 4A). In contrast, the ACE inhibitor did not increase AP-1 activity in ACE-deficient cells or in endothelial cells expressing the S1270A ACE mutant.

View larger version (33K): [in this window] [in a new window]   Figure 4. Role of AP-1 in the ramiprilat-induced increase in COX-2 promoter activity. A, Electrophoretic mobility shift assay showing the effect of ramiprilat (R, 100 nmol/L, 15 minutes) on the DNA binding activity of AP-1 in ACE-overexpressing or S1270A-overexpressing and ACE-deficient (–ACE), endothelial cells. B, ACE-overexpressing endothelial cells were transfected with the COX-2 promoter construct in the absence (Solv) and presence of AP-1 decoy (Decoy) or scrambled (Scr) oligonucleotides (0.5 µmol/L) and incubated with either solvent (C, CTL) or ramiprilat (R, 100 nmol/L) for 36 hours before the analysis of promoter activity. The bar graphs summarize data obtained in 4 to 10 different experiments; *P<0.05 vs CTL.

To directly demonstrate the involvement of AP-1 in the ramiprilat-induced upregulation of COX-2, we assessed the effects of AP-1 decoy oligonucleotides. ACE-overexpressing endothelial cells were incubated with either control oligonucleotides (AP-1 scrambled) or AP-1 decoy oligonucleotides immediately after transfection with the COX-2 promoter and before the addition of the ACE inhibitor. The ramiprilat-induced increase in COX-2 promoter activity was unaffected by scrambled oligonucleotides but was prevented by the AP-1 decoy oligonucleotides (Figure 4B).

AP-1 decoy oligonucleotides also prevented the ramiprilat-induced increase in endogenous COX-2 expression in primary cultures of human endothelial cells (Figure 5A). The ramiprilat-induced increase in COX-2 expression was accompanied by an increase in the accumulation of the stable PGI₂ metabolite, 6-keto PGF₁ in the cell supernatant. The specific COX-2 inhibitor celecoxib (1 µmol/L) attenuated the basal production of 6-keto PGF₁ in solvent-treated cells by 75±3% (n=5, P<0.01) and completely prevented the ACE inhibitor-induced increase in the autacoid so that in the presence of celecoxib 6-keto PGF₁ levels did not differ between the control and ramiprilat-treated groups (Figure 5B). Although ramiprilat also enhanced the celecoxib-sensitive generation of PGE₂ (by 20±5%, n=8, P<0.01), the absolute levels measured were much lower than those of PGI₂, which is in agreement with several previous studies.^21,22 PGI₂ production by ramiprilat-stimulated cells ranged from 6 to 12 ng/mL, whereas PGE₂ levels were between 100 and 220 pg/mL. We were unable to detect any effect of ramiprilat on the production of thromboxane A₂ (data not shown).

View larger version (34K): [in this window] [in a new window]   Figure 5. Effect of ramiprilat and AP-1 decoy oligonucleotides on COX-2 expression and activity. Primary cultures of human endothelial cells were incubated with solvent (C, CTL) or ramiprilat (R, Rami; 100 nmol/L) for 36 hours in the absence (Solv) and presence of (A) AP-1 decoy or scrambled (Scr) oligonucleotides (0.5 µmol/L) or (B) celecoxib (1 µmol/L). 6-Keto PGF1 was assayed in the cells supernatant and the bar graphs summarize the results of 3 to 4 independent experiments; *P<0.05 vs CTL, P<0.01 vs the respective Solv.

	Discussion

Top Abstract Introduction Materials and Methods Results Discussion References

 
The results of the present investigation demonstrate that the binding of the ACE inhibitor, ramipril, to ACE directly elicits a signaling cascade that results in the activation of the transcription factor AP-1 and an increase in the expression and activity of COX-2 in endothelial cells. This signaling cascade is likely to underlie the increase in PGI₂ production reported in patients treated with ACE inhibitors.^14,15

The positive effects of ACE inhibitor therapy are generally attributed to the inhibition of ACE activity and the subsequent decrease in the production of angiotensin II and the accumulation of bradykinin. However, numerous experimental studies clearly indicate that some of the effects of ACE inhibitors cannot be directly attributed to changes in the concentration of either angiotensin II or bradykinin. We have recently reported that ACE is an ectoenzyme and also possesses properties that implicate its involvement in cellular signaling. For example, the cytoplasmic tail of ACE, more specifically Ser¹²⁷⁰, is phosphorylated,¹⁷ and the binding of an ACE inhibitor to ACE increases its phosphorylation and elicits the activation of the ACE-associated kinases CK2 and JNK.¹¹ As a consequence of the activation of JNK in ACE inhibitor-treated cells, c-Jun is translocated to the nucleus and the expression of ACE is increased. The latter effect is not limited to an artificial cell culture system but can be reproduced in mice given an ACE inhibitor in the drinking water,¹¹ and has been documented in individuals treated with ACE inhibitors.^9,10 Although this evidence certainly indicates that "ACE signaling" is a relevant physiological process, it is difficult to envisage that the reported increase in ACE expression can actually contribute to the beneficial effects of ACE inhibitor therapy.

Because the expression of COX-2, like that of ACE, is reported to be regulated by mechanisms involving c-Jun²⁰ and the transcription factor AP-1,^12,20 we determined whether the activation of ACE signaling can affect the expression and activity of COX-2. We found that ramiprilat increases the expression of COX-2 in different experimental models; in primary cultures of human umbilical endothelial cells that endogenously express ACE; in a porcine aortic endothelial cell line overexpressing human somatic ACE; and in the lungs of mice treated with ramipril for 5 days. Strictly speaking, it is impossible to exclude a role for ACE inhibitor-induced alterations in the concentration of angiotensin II or bradykinin in the responses observed in the human or mouse endothelial cells. However, the fact that COX-2 expression was not enhanced in angiotensin and bradykinin receptor-deficient endothelial cell lines that do not express ACE or in cells that overexpress a nonphosphorylatable S1270A mutant certainly indicates an ACE-dependent signaling process. By interfering with the activity of JNK and AP-1, it was possible to prevent the ramiprilat-induced increase in COX-2 promoter activity and protein expression, but again only in cells expressing wild-type ACE.

At first glance, it is not evident that an increase in the expression of COX-2 can be associated with vasoprotective effects because COX-2 is still considered as the "bad" COX isoform, which is generally induced in response to an inflammatory reaction. On the basis of findings obtained in other cell systems, it has been generally assumed that COX-2 plays a detrimental role in cardiovascular homeostasis. Although it is clear that COX-2–derived thromboxanes would be expected to induce vasoconstriction and potentiate an inflammatory state, the enzyme generates mainly PGI₂ when it is expressed in endothelial cells.^23,24 We detected only low levels of PGE₂ in primary cultures of ramiprilat-treated human endothelial cells and were unable to detect thromboxane production. Further evidence that COX-2-derived PGI₂ is beneficial comes from studies reporting that selective COX-2 inhibition attenuates acetylcholine-induced vasodilatation in the forearm circulation of patients with essential hypertension²⁵ and diminishes the positive effects of ACE inhibitors on blood pressure.^26,27

Reports of a link between ACE inhibitor treatment and enhanced endothelium-dependent vasodilatation have long been linked with the increased production of PGI₂.²⁸ In fact, there are several mechanisms by which ACE inhibitors have been proposed to affect the synthesis of PGI₂. For example, because a short-term effect of ramiprilat (peaking within 10 to 60 minutes) on PGI₂ production could be abolished by the B₂ kinin receptor antagonist, icatibant, part of the response has been attributed to the activation of the B₂ kinin receptor signaling pathway.²⁹ However, such a rapid effect is more likely to be related to the activation of phospholipase A₂ enzymes in response to an ACE inhibitor-induced elevation in intracellular calcium¹⁶ than to an alteration in COX expression. Although the evidence linking some of the acute effects of ACE inhibitors with the transactivation of the B₂ kinin receptor⁵ or alterations in B₂ kinin receptor sequestration to caveolae⁶ is convincing, the long-term effects of ACE inhibitors on COX-2 expression reported here is unlikely to be related to the accumulation of bradykinin. The ACE-overexpressing cell line used to assess the effects of ramiprilat on COX-2 promoter activity does not express bradykinin receptors. A bradykinin-independent, long-term effect of ACE inhibitors on PGI₂ production has previously been reported in rats made hypertensive with a nitric oxide synthase inhibitor and treated with quinapril. In the latter study, the ACE inhibitor-induced improvement in flow-induced dilatation in small mesenteric arteries was sensitive to COX-2 inhibition, but not to the B₂ kinin receptor antagonist.³⁰ Because we have shown that ramipril increases COX-2 expression in mice in vivo, it is tempting to suggest that ACE signaling underlies the previously reported ACE inhibitor-induced increase in PGI₂ production.

Taken together, the results of the present investigation indicate that the enhanced production of PGI₂, which is a well-documented consequence of ACE inhibitory therapy, can be attributed to an ACE signaling cascade involving the phosphorylation of ACE and the activation of JNK, AP-1, and COX-2. Although it remains to be determined which additional intracellular mediators participate in the ACE signaling pathway, COX-2–derived PGI₂ may mediate effects other than/in addition to vasodilator function. There is at least circumstantial evidence supporting a role for COX-2 in other ACE-dependent responses inasmuch as the ACE inhibitor captopril was found to increase renal renin production in wild-type and COX-1^–/– mice but not in COX-2^–/– mice.^31,32

	Acknowledgments

 
The authors are indebted to Janna Enderich, Marie von Reutern, and Isabel Winter for expert technical assistance. The experimental work described in this manuscript was supported by the Deutsche Forschungsgemeinschaft (FL 364/1), the Heinrich and Fritz Riese-Stiftung, and a research grant from Aventis.

Received September 22, 2004; first decision October 13, 2004; accepted October 22, 2004.

	References

Top Abstract Introduction Materials and Methods Results Discussion References

 

1. Schartl M, Bocksch WG, Dreysse S, Beckmann S, Hunten U. Remodeling of myocardium and arteries by chronic angiotensin converting enzyme inhibition in hypertensive patients. J Hypertens. 1994; 12: S37–S42.
2. Mancini GB, Henry GC, Macaya C, O’Neill BJ, Pucillo AL, Carere RG, Wargovich TJ, Mudra H, Luscher TF, Klibaner MI, Haber HE, Uprichard AC, Pepine CJ, Pitt B. Angiotensin-converting enzyme inhibition with quinapril improves endothelial vasomotor dysfunction in patients with coronary artery disease. The TREND (Trial on Reversing ENdothelial Dysfunction) Study. Circulation. 1996; 94: 258–265.[Abstract/Free Full Text]
3. Yusuf S, Sleight P, Pogue J, Bosch J, Davies R, Dagenais G. Effects of an angiotensin-converting-enzyme inhibitor, ramipril, on cardiovascular events in high-risk patients. The Heart Outcomes Prevention Evaluation Study Investigators. N Engl J Med. 2000; 342: 145–153.[Abstract/Free Full Text]
4. Hecker M, Blaukat A, Bara AT, Müller-Esterl W, Busse R. ACE inhibitor potentiation of bradykinin-induced venoconstriction. Br J Pharmacol. 1997; 121: 1475–1481.[CrossRef][Medline] [Order article via Infotrieve]
5. Minshall RD, Tan F, Nakamura F, Rabito SF, Becker RP, Marcic B, Erdös EG. Potentiation of the actions of bradykinin by angiotensin I-converting enzyme inhibitors: the role of expressed human bradykinin B₂ receptors and angiotensin I-converting enzyme in CHO cells. Circ Res. 1997; 81: 848–856.[Abstract/Free Full Text]
6. Benzing T, Fleming I, Blaukat A, Müller-Esterl W, Busse R. Angiotensin-converting enzyme inhibitor ramiprilat interferes with the sequestration of the B₂ kinin receptor within the plasma membrane of native endothelial cells. Circulation. 1999; 99: 2034–2040.[Abstract/Free Full Text]
7. Fyhrquist F, Forslund T, Tikkanen I, Gronhagen-Riska C. Induction of angiotensin I-converting enzyme rat lung with Captopril (SQ 14225). Eur J Pharmacol. 1980; 67: 473–475.[CrossRef][Medline] [Order article via Infotrieve]
8. Costerousse O, Allegrini J, Clozel J-P, Menard J, Alhenc-Gelas F. Angiotensin I-converting enzyme inhibition but not angiotensin II suppression alters angiotensin I-converting enzyme gene expression in vessels and epithelia. J Pharmacol Exp Ther. 1998; 284: 1180–1187.[Abstract/Free Full Text]
9. Boomsma F, de Bruyn JH, Derkx FH, Schalekamp MA. Opposite effects of captopril on angiotensin I-converting enzyme ’activity’ and ’concentration’; relation between enzyme inhibition and long-term blood pressure response. Clin Sci (Lond). 1981; 60: 491–498.[Medline] [Order article via Infotrieve]
10. Sassano P, Chatellier G, Billaud E, Alhenc-Gelas F, Corvol P, Menard J. Treatment of mild to moderate hypertension with or without the converting enzyme inhibitor enalapril. Results of a six-month double- blind trial. Am J Med. 1987; 83: 227–235.[CrossRef][Medline] [Order article via Infotrieve]
11. Kohlstedt K, Brandes RP, Muller-Esterl W, Busse R, Fleming I. Angiotensin-converting enzyme is involved in outside-in signaling in endothelial cells. Circ Res. 2004; 94: 60–67.[Abstract/Free Full Text]
12. Eyries M, Agrapart M, Alonso A, Soubrier F. Phorbol ester induction of angiotensin-converting enzyme transcription is mediated by Egr-1 and AP-1 in human endothelial cells via ERK1/2 pathway. Circ Res. 2002; 91: 899–906.[Abstract/Free Full Text]
13. Abulencia JP, Gaspard R, Healy ZR, Gaarde WA, Quackenbush J, Konstantopoulos K. Shear-induced cyclooxygenase-2 via a JNK2/c-Jun-dependent pathway regulates prostaglandin receptor expression in chondrocytic cells. J Biol Chem. 2003; 278: 28388.[Abstract/Free Full Text]
14. Kohno M, Yokokawa K, Minami M, Yasunari K, Maeda K, Kano H, Hanehira T, Yoshikawa J. Plasma levels of nitric oxide and related vasoactive factors following long-term treatment with angiotensin-converting enzyme inhibitor in patients with essential hypertension. Metabolism. 1999; 48: 1256–1259.[CrossRef][Medline] [Order article via Infotrieve]
15. Torsello A, Locatelli V, Cella SG, Sanguini AM, Berti F. Moexipril and quinapril inhibition of tissue angiotensin-converting enzyme activity in the rat: evidence for direct effects in heart, lung and kidney and stimulation of prostacyclin generation. J Endocrinol Invest. 2003; 26: 79–83.[Medline] [Order article via Infotrieve]
16. Busse R, Lamontagne D. Endothelium-derived bradykinin is responsible for the increase in calcium produced by angiotensin-converting enzyme inhibitors in human endothelial cells. Naunyn Schmiedebergs Arch Pharmacol. 1991; 344: 126–129.[Medline] [Order article via Infotrieve]
17. Kohlstedt K, Shoghi F, Müller-Esterl W, Busse R, Fleming I. CK2 phosphorylates the angiotensin-converting enzyme and regulates its retention in the endothelial cell plasma membrane. Circ Res. 2002; 91: 749–756.[Abstract/Free Full Text]
18. Potente M, Michaelis UR, Fisslthaler B, Busse R, Fleming I. Cytochrome P450 2C9-induced endothelial cell proliferation involves induction of mitogen-activated protein (MAP) kinase phosphatase-1, inhibition of the c-Jun N-terminal kinase, and up-regulation of cyclin D1. J Biol Chem. 2002; 277: 15671–15676.[Abstract/Free Full Text]
19. Fisslthaler B, Benzing T, Busse R, Fleming I. Insulin enhances the expression of the endothelial nitric oxide synthase in native endothelial cells: a dual role for Akt and AP-1. Nitric Oxide. 2003; 8: 253–261.[CrossRef][Medline] [Order article via Infotrieve]
20. Smith WL, DeWitt DL, Garavito RM. Cyclooxygenases: structural, cellular, and molecular biology. Annual Review of Biochemistry. 2000; 69: 145–182.[CrossRef][Medline] [Order article via Infotrieve]
21. Busse R, Förstermann U, Matsuda H, Pohl U. The role of prostaglandins in the endothelium-mediated vasodilatory response to hypoxia. Pflugers Arch Eur J Physiol. 1984; 401: 77–83.[CrossRef][Medline] [Order article via Infotrieve]
22. Satoh H, Satoh S. Prostaglandin E₂ and I₂ production in isolated dog renal arteries in the absence or presence of vascular endothelial cells. Biochem Biophys Res Commun. 1984; 118: 873–876.[CrossRef][Medline] [Order article via Infotrieve]
23. Catella-Lawson F, McAdam B, Morrison BW, Kapoor S, Kujubu D, Antes L, Lasseter KC, Quan H, Gertz BJ, Fitzgerald GA. Effects of specific inhibition of cyclooxygenase-2 on sodium balance, hemodynamics, and vasoactive eicosanoids. J Pharmacol Exp Ther. 1999; 289: 735–741.[Abstract/Free Full Text]
24. McAdam BF, Catella-Lawson F, Mardini IA, Kapoor S, Lawson JA, Fitzgerald GA. Systemic biosynthesis of prostacyclin by cyclooxygenase (COX)-2: the human pharmacology of a selective inhibitor of COX-2. Proc Natl Acad Sci U S A. 1999; 96: 272–277.[Abstract/Free Full Text]
25. Bulut D, Liaghat S, Hanefeld C, Koll R, Miebach T, Mugge A. Selective cyclo-oxygenase-2 inhibition with parecoxib acutely impairs endothelium-dependent vasodilatation in patients with essential hypertension. J Hypertens. 2003; 21: 1663–1667.[CrossRef][Medline] [Order article via Infotrieve]
26. Izhar M, Alausa T, Folker A, Hung E, Bakris GL. Effects of COX inhibition on blood pressure and kidney function in ACE inhibitor-treated blacks and hispanics. Hypertension. 2004; 43: 573–577.[Abstract/Free Full Text]
27. Stollberger C, Finsterer J. Nonsteroidal anti-inflammatory drugs in patients with cardio- or cerebrovascular disorders. Z Kardiol. 2003; 92: 721–729.[CrossRef][Medline] [Order article via Infotrieve]
28. Nakagawa M, Sawada S, Toyoda T, Takamatsu H, Tsuji H, Ijichi H. Regulation of prostacyclin generation by angiotensin converting enzyme related substances in cultured human vascular endothelial cells. Clin Exp Hypertens A. 1987; 9: 405–408.[Medline] [Order article via Infotrieve]
29. Wiemer G, Schölkens BA, Becker RHA, Busse R. Ramiprilat enhances endothelial autacoid formation by inhibiting breakdown of endothelium-derived bradykinin. Hypertension. 1991; 18: 558–563.[Abstract/Free Full Text]
30. Henrion D, Dechaux E, Dowell FJ, Maclour J, Samuel JL, Levy BI, Michel JB. Alteration of flow-induced dilatation in mesenteric resistance arteries of L-NAME treated rats and its partial association with induction of cyclo-oxygenase-2. Br J Pharmacol. 1997; 121: 83–90.[Medline] [Order article via Infotrieve]
31. Cheng HF, Wang JL, Zhang MZ, Wang SW, McKanna JA, Harris RC. Genetic deletion of COX-2 prevents increased renin expression in response to ACE inhibition. Am J Physiol Renal Physiol. 2001; 280: F449–F456.[Abstract/Free Full Text]
32. Cheng HF, Wang SW, Zhang MZ, McKanna JA, Breyer R, Harris RC. Prostaglandins that increase renin production in response to ACE inhibition are not derived from cyclooxygenase-1. Am J Physiol Regul Integr Comp Physiol. 2002; 283: R638–R646.[Abstract/Free Full Text]

This article has been cited by other articles:

				C. Kojima, A. Kawakami, T. Takei, K. Nitta, and M. Yoshida Angiotensin-Converting Enzyme Inhibitor Attenuates Monocyte Adhesion to Vascular Endothelium through Modulation of Intracellular Zinc J. Pharmacol. Exp. Ther., December 1, 2007; 323(3): 855 - 860. [Abstract] [Full Text] [PDF]

				Z. Chen, P. A. Deddish, R. D. Minshall, R. P. Becker, E. G. Erdos, and F. Tan Human ACE and bradykinin B2 receptors form a complex at the plasma membrane FASEB J, November 1, 2006; 20(13): 2261 - 2270. [Abstract] [Full Text] [PDF]

				S. Donnini, R. Solito, A. Giachetti, H. J. Granger, M. Ziche, and L. Morbidelli Fibroblast Growth Factor-2 Mediates Angiotensin-Converting Enzyme Inhibitor-Induced Angiogenesis in Coronary Endothelium J. Pharmacol. Exp. Ther., November 1, 2006; 319(2): 515 - 522. [Abstract] [Full Text] [PDF]

				M. Paul, A. Poyan Mehr, and R. Kreutz Physiology of local Renin-Angiotensin systems. Physiol Rev, July 1, 2006; 86(3): 747 - 803. [Abstract] [Full Text] [PDF]

				K. Kohlstedt, C. Gershome, M. Friedrich, W. Muller-Esterl, F. Alhenc-Gelas, R. Busse, and I. Fleming Angiotensin-Converting Enzyme (ACE) Dimerization Is the Initial Step in the ACE Inhibitor-Induced ACE Signaling Cascade in Endothelial Cells Mol. Pharmacol., May 1, 2006; 69(5): 1725 - 1732. [Abstract] [Full Text] [PDF]

				I. Fleming Signaling by the Angiotensin-Converting Enzyme Circ. Res., April 14, 2006; 98(7): 887 - 896. [Abstract] [Full Text] [PDF]

				D. Biyashev, F. Tan, Z. Chen, K. Zhang, P. A. Deddish, E. G. Erdos, and C. Hecquet Kallikrein activates bradykinin B2 receptors in absence of kininogen Am J Physiol Heart Circ Physiol, March 1, 2006; 290(3): H1244 - H1250. [Abstract] [Full Text] [PDF]

				Z. Chen, F. Tan, E. G. Erdos, and P. A. Deddish Hydrolysis of Angiotensin Peptides by Human Angiotensin I-Converting Enzyme and the Resensitization of B2 Kinin Receptors Hypertension, December 1, 2005; 46(6): 1368 - 1373. [Abstract] [Full Text] [PDF]

				M. L. Hemming and D. J. Selkoe Amyloid {beta}-Protein Is Degraded by Cellular Angiotensin-converting Enzyme (ACE) and Elevated by an ACE Inhibitor J. Biol. Chem., November 11, 2005; 280(45): 37644 - 37650. [Abstract] [Full Text] [PDF]

				P. REDELINGHUYS, A. T. NCHINDA, and E. D. STURROCK Development of Domain-Selective Angiotensin I-Converting Enzyme Inhibitors Ann. N.Y. Acad. Sci., November 1, 2005; 1056(1): 160 - 175. [Abstract] [Full Text] [PDF]

				O. A. Carretero Novel mechanism of action of ACE and its inhibitors Am J Physiol Heart Circ Physiol, November 1, 2005; 289(5): H1796 - H1797. [Full Text] [PDF]

				I. Fleming, K. Kohlstedt, and R. Busse New fACEs to the Renin-Angiotensin System Physiology, April 1, 2005; 20(2): 91 - 95. [Abstract] [Full Text] [PDF]

This Article Abstract Full Text (PDF) All Versions of this Article: 45/1/126    most recent 01.HYP.0000150159.48992.11v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Kohlstedt, K. Articles by Fleming, I. Search for Related Content PubMed PubMed Citation Articles by Kohlstedt, K. Articles by Fleming, I. Pubmed/NCBI databases Gene GEO Profiles HomoloGene UniGene Compound via MeSH Substance via MeSH Hazardous Substances DB CELECOXIB Related Collections Cardiovascular Pharmacology Other Vascular biology ACE/Angiotension receptors Cell signalling/signal transduction Gene expression