doi: 10.1161/hq0202.104123
© 2002 American Heart Association, Inc.
Vascular Biology |
Prostacyclin Synthase Gene Transfer Modulates Cyclooxygenase-2–Derived Prostanoid Synthesis and Inhibits Neointimal Formation in Rat Balloon-Injured Arteries
From Internal Medicine II (M.Y., K.O., M.H., H.M., T.H.), Physiology II (K.N.), and the Department of Health Sciences (T.I.), Nagoya University School of Medicine, Nagoya, Japan, and the Departments of Surgery and Pathology (Y.N.), Children’s Hospital, Harvard Medical School, Boston, Mass.
Correspondence to Kenji Okumura, MD, Internal Medicine II, Nagoya University School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8550, Japan. E-mail kenji{at}med.nagoya-u.ac.jp
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Abstract |
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Previous studies have shown that prostacyclin (PGI2) synthase (PCS) gene transfer inhibits neointimal formation in balloon-injured arteries. However, the role of each cyclooxygenase (COX) isoform in this healing mechanism remains unknown. We hypothesized that overexpression of PCS may modulate COX-2–mediated prostaglandin (PG) metabolism. That is to say, excessive PGH2 derived from COX-2 after balloon injury may be converted into PGI2 rather than PGE2 or thromboxane (TX) A2 by overexpressed PCS. We examined the expression of COX isoforms and evaluated the role of COX-2 with regard to the effects of PCS gene transfer by using 4-(4-cyclohexyl-2-methyloxazol-5-yl)-2-fluorobenzenesulfonamide (JTE-522), a selective COX-2 inhibitor. Rats were divided into 4 groups in conjunction with PCS gene transfer and JTE-522 treatment. The PCS gene (30 µg) was transfected into rat balloon-injured arteries by a lipotransfection method. JTE-522 (30 mg/kg per day) was administered for 14 days after balloon injury. Immunohistochemical analysis demonstrated marked COX-2 expression on the neointima. PCS gene transfer markedly inhibited neointimal formation, but JTE-522 reversed this beneficial effect. PCS gene transfer augmented PGI2 production and decreased PGE2 production without affecting TXA2 production, but JTE-522 inhibited this increase in PGI2 production. In conclusion, PCS gene transfer modulated COX-2–mediated prostanoid synthesis and inhibited neointimal formation after balloon injury.
Key Words: prostacyclin • cyclooxygenase-2 • gene therapy • restenosis • balloon injury
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Introduction |
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Cyclooxygenase (COX) is the key enzyme that regulates the amount and duration of prostaglandin (PG) production in vessels.1,2 Recently, the role of COX in cardiovascular diseases such as atherosclerosis has been studied extensively. COX converts arachidonic acid into PGG2 and then into PGH2. PGH2 is metabolized by specific isomerases to prostanoids such as prostacyclin (PGI2), thromboxane (TX) A2, and PGE2, which exert a variety of biological actions involved in the maintenance of vascular homeostasis.3 There are 2 isoforms of the enzyme, COX-1 and COX-2, and they are encoded by distinct genes.4,5 COX-1 is constitutively expressed in most tissues, whereas COX-2 is an inducible isoform and almost undetectable in physiological conditions.6 COX-2 is induced by interleukin-1ß, tumor necrosis factor-
, bacterial lipopolysaccharide, growth factors, phorbol esters, and other agents.7 In normal vessels, the endothelium is enriched with COX activity,8 and the constitutive isoform of COX (COX-1) determines PG production in the endothelium.9 Once the barrier function of the endothelium is disrupted by an injury, such as angioplasty, COX-1 activity may decrease, and in turn, to compensate for the loss of COX-1 function, COX-2 expression is rapidly induced to an excessive level, resulting in an imbalance of PGs among PGH2, PGE2, PGI2, and TXA2. In that case, the vessel wall tends to be vasoconstrictive and thrombogenic, which might, in turn, lead to atherosclerosis.10
PGI2 is generated by vessel endothelium and vascular smooth muscle cells (VSMCs).11 PGI2 is a potential vasodilator,12 which inhibits smooth muscle cell proliferation13 and platelet aggregation via cAMP14 and also modulates cholesterol turnover.15 Thus, PGI2 may play an important role in modulating the vascular response to injury. We have recently shown that prostacyclin synthase (PCS) gene transfer inhibits neointimal formation in rat balloon-injured arteries16,17; however, the role of each COX isoform in this healing mechanism after arterial injury remains unknown. Pritchard et al18 have reported that the smooth muscle cells of arteries express COX-2 mRNA and COX-2 protein after balloon injury. Moreover, there is evidence that COX-2 is expressed on VSMCs in human atherosclerotic lesions.19–21 Previous studies have indicated that there may be a specific link between the accumulation of PGE2 and the induction of COX-2.22,23 We hypothesized that COX-2–derived excessive PGH2 could be converted into PGI2 rather than PGE2 and TXA2 by the overexpression of PCS. We carried out immunohistochemical analysis to clarify the distribution of COX isoforms in rat arterial walls after balloon injury and evaluated the involvement of COX-2 by using 4-(4-cyclohexyl-2-methyloxazol-5-yl)-2-fluorobenzenesulfonamide (JTE-522), a selective COX-2 inhibitor, after PCS gene transfer.
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Methods |
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Preparation of PCS Plasmid DNA
The plasmids encoding the PCS gene used in the present study have been described in detail previously.16,17 Briefly, a polymerase chain reaction–amplified PCS gene was cloned into pTarget (Promega), a cytomegalovirus (CMV) enhancer/promoter and simian virus 40 polyadenylation signal–driven mammalian expression vector, yielding pCMV-PCS. The functional character of these plasmids, such as protein expression level in vitro, has been confirmed in our previous reports.16,17
Gene Delivery In Vivo and Inhibition of COX-2
Male Sprague-Dawley rats weighing 350 to 400 g were obtained from Japan SLC, Inc (Nagoya, Japan). We followed the National Institutes of Health guidelines regarding care and use of the animals. The in vivo gene transfer into rat carotid arteries was performed as previously described.16,17 Rats were anesthetized with sodium pentobarbital (50 mg/kg IP). After a midcervical incision, the right common carotid artery and its bifurcation were exposed. To prevent acute thrombosis during the procedure, heparin sodium (200 IU/kg) was intravenously injected 5 minutes before the balloon injury. The right common carotid artery was balloon-injured 3 times with a 2F Fogarty catheter (Baxter Healthcare) inserted through the external carotid artery as previously described by Clowes et al.24 To attain a constant degree of vessel wall injury for each of the animals, we kept the diameter of the balloon and the resistance during withdrawal constant and the same for each of the animals. The sham operation involved simple ligation of the right external carotid arteries without balloon injury. Two vascular clips were placed at the distal end and in the middle of the injured arterial segment. A 24-gauge cannula was introduced into the common carotid artery via the external carotid artery. The lumen of the injured segment between the 2 clips was washed with PBS. pCMV-PCS (30 µg) with 100 µL Lipofectamine Plus reagent (GIBCO-BRL) in a total volume of 300 µL was instilled into the lumen for 30 minutes, and then the catheter was carefully withdrawn, and the blood flow was restored. After confirmation of the carotid arterial flow, the wound was sutured. All procedures were performed under sterile conditions. In a prior study, the extent of endothelial denudation was confirmed at 2 days after balloon injury by Evans blue staining. We confirmed that the present plasmid vector system could transfer target genes into the injured arteries by using a plasmid carrying the lacZ gene (pCMV-lacZ) based on X-Gal staining, and the transgene efficiency was 8.5% in our previous reports.16,17 JTE-522 was suspended in 0.5% carboxymethylcellulose sodium (CMC-Na) solution. JTE-522 (30 mg/kg) or vehicle (0.5% CMC-Na) was administered orally at a volume of 5 mL/kg with gastric gavage starting 30 minutes before the injury and continuing for 14 days. The balloon-injured rats were divided into 4 groups in conjunction with PCS gene transfer and COX-2 inhibitor treatment as follows: (1) vehicle (injured group), (2) pCMV-PCS plus vehicle (pPCS group), (3) JTE-522 (JTE-522 group), and (4) pCMV-PCS plus JTE-522 (pPCS+JTE-522 group). All groups had 6 rats. Systolic blood pressure and heart rate were measured before and 14 days after balloon injury by the tail-cuff method with a sphygmomanometer (BP98A, Softron). The urinary level of 6-keto-PGF1 was measured with a radioimmunoassay kit (NEN).
Immunohistochemistry
To evaluate COX-1 and COX-2 expression in balloon-injured arteries (n=6) and uninjured arteries from sham-operated animals (n=6), the rats were killed at 3, 7, and 14 days after balloon injury, and the right carotid arteries were perfusion-fixed with 2.5% phosphate-buffered glutaraldehyde and harvested under anesthesia with sodium pentobarbital. Immunohistochemical labeling was carried out on adjacent tissue sections. The excised carotid arteries were paraffin-embedded, cut into 4-µm cross sections, and mounted on glass slides. These sections were incubated in 0.3% hydrogen peroxide methanol for 30 minutes to block endogenous peroxidase activity. The nonspecific binding of rabbit serum was prevented by preincubating the sections with 0.1% normal goat serum. The sections were sequentially incubated at 4°C overnight with polyclonal rabbit anti-murine COX-1 antibody (catalogue No. 160109, Cayman Chemical Co)25 at a concentration of 1:200 or polyclonal rabbit anti-murine COX-2 antibody (catalogue No. 160106, Cayman Chemical Co)26 at a concentration of 1:200. After they were washed with PBS, the sections were incubated with biotinylated goat anti-rabbit IgG-conjugated horseradish peroxidase (Dako Japan Co) for an additional 60 minutes at room temperature. Each incubation was followed by a wash in PBS. Staining was visualized with chromogen, 0.06% 3,3'-diaminobenzidine/5% hydrogen peroxide in 0.05 mol/L Tris-HCl (pH 7.6), and hematoxylin for a counterstain. Control sections were incubated with nonimmune rabbit IgG at a concentration of 1:200.
Prostanoid Production in Rat Carotid Artery
The injured carotid arteries of each group were resected 7 days after balloon injury. The arteries were cut into 5-mm lengths, washed with PBS, and incubated in 1 mL of PBS (pH 7.4) at 37°C for 45 minutes. The levels of 6-keto-PGF1, TXB2, and PGE2 in the medium were measured to evaluate local PGI2, TXA2, and PGE2 production, respectively, with radioimmunoassay kits (NEN). Moreover, we similarly examined the effect of COX-1 and COX-2 blockade in the injured and PCS gene-transfected arteries (n=6 each) by using indomethacin (Sigma), a nonselective COX inhibitor. Indomethacin (5 mg/kg) was suspended in 0.5% CMC-Na solution and administered orally at a volume of 5 mL/kg with gastric gavage once daily for 7 days after balloon injury.
Quantification of Neointimal Formation
Fourteen days after balloon injury, 6 cross sections from each carotid artery were stained with hematoxylin and eosin and examined morphometrically with a computerized digital image analysis system (NIH image) in a blind manner. The areas within the external elastic lamina (EEL area), the internal elastic lamina (IEL area), and the luminal area were measured. Other areas were calculated as follows: medial area=EEL area-IEL area; neointimal area=IEL area-luminal area; neointima-to-media (I/M) ratio=neointimal area/medial area.
Statistical Analysis
All data are expressed as the mean±SEM. Statistical analysis for multiple comparisons among the groups used 1-way ANOVA followed by the Bonferroni test. A value of P<0.05 was considered statistically significant.
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Results |
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COX Isoform Expression in Rat Balloon-Injured and Uninjured Arterial Vessel Wall
Fourteen days after balloon injury, sections of the injured and uninjured arterial segments were analyzed for COX-1 and COX-2 protein by immunohistochemical analysis (n=6 each). In the balloon-injured arteries, there was diffuse expression of COX-2 protein on the neointima and adventitia (Figure 1D and 1F), whereas COX-1 expression was not detectable on the neointima (Figure 1C and 1E). Interestingly, COX-1 and COX-2 colocalized with regenerated endothelial cells (inset, Figure 1E and 1F). In contrast, in the uninjured control arteries, there was constitutive expression of COX-1 protein in the endothelium and adventitia (Figure 1A), whereas there was no detectable COX-2 except in the adventitia (Figure 1B).
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Time Course of COX-1 and COX-2 Expression
We examined the time course of COX-1 and COX-2 expression after balloon injury by immunohistochemistry. COX-2 was expressed in the exposed subendothelium and in the adventitia by 3 days after injury (Figure 2B), it was clearly evident in the neointimal and adventitial areas by 7 days (Figure 2C), and it was widely expressed in the same areas by 14 days (Figure 2D). However, COX-1 expression was not detectable in the neointima except for the regenerated endothelium at any time after injury.
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Relative Influence of PCS Gene Transfer and COX-1 and COX-2 Blockade on Prostanoid Production in Rat Carotid Arteries
We evaluated the production of PGI2 in the arterial vessels by determining the 6-keto-PGF1 levels 7 days after injury and found the levels to be significantly lower in balloon-injured vessels than the uninjured vessels. In contrast, the levels were significantly higher in pCMV-PCS–transfected vessels than in the injured vessels. The PCS gene transfer restored the 6-keto-PGF1 levels to values even higher than those of the uninjured vessels; however, this effect of pCMV-PCS gene transfer was markedly suppressed with either indomethacin or JTE-522 (n=6 in each group, Figure 3A). We evaluated TXA2, the counterpart of derivatives from PGH2 in the same samples, by determining the levels of TXB2. The TXB2 production levels were higher in the injured vessels than in the uninjured vessels, whereas there was no significant difference in the TXB2 levels between the injured and pCMV-PCS–transfected vessels. Thus, balloon-injured carotid arteries transfected with the PCS gene were able to produce a larger amount of PGI2 without any changes in their ability to produce TXA2. JTE-522 had no effect on TXB2 production. In contrast, indomethacin markedly suppressed TXB2 production (n=6 in each group, Figure 3B). Moreover, PGE2 levels, which are thought to reflect COX-2 activity in tissue,22,23 were measured in the same samples. There was no significant difference in the PGE2 levels between the injured and uninjured vessels, whereas the levels in pCMV-PCS–transfected vessels were significantly lower than the levels in the injured vessels. JTE-522 and indomethacin also markedly suppressed PGE2 production levels (n=6 in each group, Figure 3C).
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P<0.05 vs uninjured; and 
P<0.01 vs pPCS.
Systemic Effects of PCS Gene Transfer and JTE-522 Administration
Fourteen days after injury, systolic blood pressures for the test rats were 129±7, 124±2, 129±4, and 129±3 mm Hg in the injured, pPCS, JTE-522, and pPCS+JTE-522 groups, respectively (n=6 each, P=NS). Regarding heart rate, no significant difference was observed among the 4 groups (383±18, 378±13, 391±14, 372±8 bpm for the injured, pPCS, JTE-522, and pPCS+JTE-522 groups, respectively). The baseline levels of urinary 6-keto-PGF1 were not significantly different among the 4 groups. Seven days after balloon injury, the urinary levels of 6-keto-PGF1 did not significantly change in any of the groups (data not shown). Thus, local delivery of pCMV-PCS to the carotid artery and systemic administration of JTE-522 had no effect on the systemic production of 6-keto-PGF1.
Effects of PCS Gene Transfer and JTE-522 on Neointimal Formation
Figure 4 shows representative histological photomicrography of common carotid arteries 14 days after the injury. Figure 5 shows the morphometric analysis of these arteries. Morphometric analysis showed that the neointimal area in the pPCS group was reduced by 47.4% and that the I/M ratio was reduced by 45.5%, which provided larger luminal areas compared with the injured group 14 days after the injury. However, these beneficial effects of PCS gene transfer were reversed by JTE-522. There was no significant difference in the EEL area or medial area among the 4 groups.
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Discussion |
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In the present study, we have shown that there is a marked expression of COX-2 on the neointima and adventitia in balloon-injured rat carotid arteries, whereas there is less expression of COX-1 on the neointima as observed during immunohistochemical analysis. Increased COX-2 expression was observed by 3 days after injury and was sustained up to 14 days. Furthermore, we observed significant inhibition of neointimal formation (47.4% and 45.5% reduction in the neointimal area and the I/M ratio, respectively) at 14 days after injury by single PCS gene transfer, but this beneficial effect was reversed by the COX-2 inhibitor in this animal model of restenosis.
We used JTE-522 as a selective COX-2 inhibitor. JTE-522 is selective for COX-2 at a daily dose of 0.3 to 30 mg/kg, and it does not cause severe gastric lesions at oral doses up to 300 mg/kg in rats.27 We administered JTE-522 (30 mg/kg per day) orally with gastric gavage for 14 days. This relatively high dose of JTE-522 was selected because optimal inhibition of COX-2 in the vessel walls was insufficient with a dose <10 mg/kg per day in our prior study. At this dose, we did not observe any systemic side effects derived from the administration of JTE-522.
We have shown the time course of COX isoform expression and localization in balloon-injured arteries by immunohistochemical analysis. Pritchard et al18 have reported that smooth muscle cells of rat arteries express COX-2 mRNA and COX-2 protein during 14 days after balloon injury by Northern and Western blot analysis. We could demonstrate COX-2 expression in the neointima and adventitia up to 14 days after balloon injury. The present results are consistent with those of Pritchard et al.
We did not observe a significant reduction in neointimal formation after arterial injury by JTE-522 at a dose of 30 mg/kg per day. Although we obtained an inhibition of neointimal formation after balloon injury by PCS gene transfer, this effect was completely reversed by JTE-522 treatment. These findings demonstrate that the effects of PCS gene transfer in injured arteries are functionally concerned with COX-2 expression. In terms of PG production, PGI2 production decreased significantly after balloon injury, whereas PCS gene transfer increased to more than the levels found in uninjured vessels. This effect of PCS gene transfer was reversed by either JTE-522 or indomethacin to the levels found in balloon-injured arteries (Figure 3A). PGI2 synthesis in the balloon-injured arteries that were transfected with the PCS gene appears to be tightly coupled with the COX-2 protein expression that increased after balloon injury in vivo. PGE2 enhances platelet aggregation28,29 and chemotaxis of leukocytes, it increases vascular permeability,30,31 and it can inhibit in vitro cholesterol ester hydrolase activity, thus leading to lipid deposition in the arterial walls.32 Therefore, an increased release of PGE2 may promote the development of atherosclerosis.10 PGE2 production, which is thought to reflect COX-2 activity,22,23 was not significantly changed by balloon injury. JTE-522 and indomethacin markedly suppressed PGE2 production in the injured arteries (Figure 3C), whereas JTE-522 had no effect on PGE2 production in the uninjured arteries in our preliminary study (data not shown). These results suggest that in normal vessels, COX-1 contributes to PGE2 synthesis, whereas COX-2 mainly contributes in the injured arteries. Thus, the induction of COX-2 in the injured arteries may represent a compensatory mechanism that ensures the release of PGE2 when the endothelium is damaged. In the present study, we found that PGE2 production was significantly reduced by PCS gene transfer after balloon injury. This phenomenon may contribute to the reduction of neointimal formation after arterial injury. Although TXA2 is mostly generated by platelets in normal vessels,33 it is produced in only very small amounts by endothelium and VSMCs.34 In the present study, TXA2 synthesis was significantly increased after balloon injury. Indomethacin markedly inhibited the increase of TXA2 production in the injured vessels, whereas JTE-522 had little effect on it (Figure 3B). These findings suggest that in the injured arteries, COX-1 mainly contributes to the increase in TXA2 synthesis. However, platelets that had adhered to the injured vessel walls may be responsible for this increase of TXA2 production in the injured arteries, because platelet COX-1 is the major source of TXA2.33 The balloon-injured arteries transfected with the PCS gene produced higher levels of PGI2 without affecting TXA2 production, as we have shown previously.16,17 Because TXA2 is a counterpart of PGI2 and facilitates platelet aggregation and VSMC proliferation partly via stimulation of growth factors such as platelet-derived growth factor,35,36 a selective augmentation of PGI2 production caused by PCS gene transfer might contribute to the reduction of neointimal formation. Furthermore, overproduction of PGH2 derived from COX-2 may cause vascular contraction and platelet aggregation.37,38 In contrast, it is reported that COX-2 induction is involved in the acceleration of wound healing, partly via angiogenesis.39,40 In our system, PCS gene transfer may effectively augment PGI2 synthesis by converting PGH2 into PGI2 without interfering with COX-2 induction. These points might be more beneficial than other genetic manipulation of the COX pathway. Finally, we propose the interesting hypothesis that the balloon injury and PCS gene transfer may modulate local PG metabolism distal to the COX pathway in the vessel wall, as shown in Figure 6.
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In conclusion, we demonstrated that the local delivery of PCS gene markedly inhibited neointimal formation in an arterial balloon-injury model. Moreover, we showed that in balloon-injured arteries, COX-2–derived PGH2 could be converted into PGI2 rather than PGE2 or TXA2 by overexpressed PCS. Manipulation of PG metabolism distal to the COX pathway might affect the development of neointimal formation even more beneficially than COX-2 inhibition and could be the basis for novel approaches to the prevention of restenosis after angioplasty. Further investigations should focus on the roles of prostanoids in postangioplastic restenosis.
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Acknowledgments |
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This work was in part supported by grants from the Ministry of Education, Science, and Culture of Japan (No. 12670658 to H. Matsui). JTE-522 was kindly donated by Japan Tobacco Inc (Tokyo, Japan). The authors thank Masafumi Ito for his excellent technical assistance with the immunohistochemical analysis and helpful discussions. This study was performed mainly at the Institute for Laboratory Animal Research, Nagoya University.
Received November 11, 2001; accepted December 5, 2001.
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References |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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1. Sanduja SK, Tsai A-L, Matijevic-Aleksic N, Wu KK. Kinetics of prostacyclin synthesis in a PGHS-1 overexpressed endothelial cell. Am J Physiol. 1994; 267: C1459–C1466.[Medline] [Order article via Infotrieve]
2. Kent RS, Diedrich SL, Whorton R. Regulation of vascular prostaglandin synthesis by metabolites of arachidonic acid in perfused rabbit aorta. J Clin Invest. 1983; 72: 455–465.[Medline] [Order article via Infotrieve]
3.
Smith WL, Garavito RM, DeWitt DL. Prostaglandin endoperoxide H synthases (cyclo-oxygenases)-1 and -2. J Biol Chem. 1996; 271: 33157–33160.
4.
Hla T, Neilson K. Human cyclooxygenase-2 cDNA. Proc Natl Acad Sci U S A. 1992; 89: 7384–7388.
5.
Xie WL, Chipman JG, Robertson DL, Erikson RL, Simmons DL. Expression of a mitogen-responsive gene encoding prostaglandin synthase is regulated by mRNA splicing. Proc Natl Acad Sci U S A. 1991; 88: 2692–2696.
6. O’Neill G, Ford-Hutchinson AW. Expression of mRNA for cyclooxygenase-1 and cyclooxygenase-2 in human tissues. FEBS Lett. 1993; 330: 156–160.[Medline] [Order article via Infotrieve]
7. Michell JA, Larkin SW, Williams TJ. Cyclo-oxygenase-2: regulation and relevance in inflammation. Biochemical Pharmacol. 1995; 50: 1535–1542.[CrossRef][Medline] [Order article via Infotrieve]
8. Moncada S, Gryglewski R, Bunting S, Vane JR. An enzyme isolated from arteries transforms prostaglandin endoperoxides to an unstable substance that inhibits platelet aggregation. Nature. 1976; 263: 663–665.[CrossRef][Medline] [Order article via Infotrieve]
9.
Mitchell JA, Akarasereenont P, Thiemermann C, Flower RJ, Vane JR. Selectivity of nonsteroidal antiinflammatory drugs as inhibitors of constitutive and inducible cyclooxygenase. Proc Natl Acad Sci U S A. 1993; 90: 11693–11697.
10.
Rolland PH, Jouve R, Pellegrin E, Mercier C, Serradimigni A. Alteration in prostacyclin and prostaglandin E2 production: correlation with changes in human aortic atherosclerotic disease. Arteriosclerosis. 1984; 4: 70–78.
11. Smith WL, De Witt DL, Allen ML. Biochemical distribution of PGI2 synthase antigen in smooth muscle cells. J Biol Chem. 1983; 258: 4922–4926.
12. Lam JYT, Chesebro JH, Badimon L, Fuster V, Exogenous prostacyclin decreases vasoconstriction but not platelet thrombus deposition after arterial injury. J Am Coll Cardiol. 1993; 21: 488–492.[Abstract]
13. Jones DA, Benjamin CW, Linseman DA. Activation of thromboxane and prostacyclin receptors elicits opposing effects on vascular smooth muscle cell growth and mitogen-activated protein kinase signaling cascades. Mol Pharmacol. 1995; 48: 890–896.[Abstract]
14. Moncada S, Vane JR. Pharmacology and endogenous roles of prostaglandin endoperoxides, thromboxane A2 and prostacyclin. Pharmacol Rev. 1979; 30: 293–331.
15. Isogaya M, Yamada N, Noike H, Ueno Y, Kumagai H, Ochi Y, Okazaki S, Nishio S. Inhibition of restenosis by beraprost sodium (a prostaglandin I2 analogue) in the atherosclerotic rabbit artery after angioplasty. J Cardiovasc Pharmacol. 1995; 25: 947–952.[Medline] [Order article via Infotrieve]
16.
Numaguchi Y, Naruse K, Harada M, Osanai H, Mokuno S, Murase K, Matsui H, Toki Y, Ito T, Okumura K, et al. Prostacyclin synthase gene transfer accelerates reendothelialization and inhibits neointimal formation in rat carotid arteries after balloon injury. Arterioscler Thromb Vasc Biol. 1999; 19: 727–733.
17.
Harada M, Toki Y, Numaguchi Y, Osanai H, Ito T, Okumura K, Hayakawa T. Prostacyclin synthase gene transfer inhibits neointimal formation in rat balloon-injured arteries without bleeding complications. Cardiovasc Res. 1999; 43: 481–491.
18.
Pritchard KA Jr, O’Banion MK, Miano JM, Vlasic N, Bhatia UG, Young DA, Stemerman MB. Induction of cyclooxygenase-2 in rat vascular smooth muscle cells in vitro and in vivo. J Biol Chem. 1994; 269: 8504–8509.
19.
Baker CS, Hall RJ, Evans TJ, Pomerance A, Maclouf J, Creminon C, Yacoub MH, Polak JM. Cyclooxygenase-2 is widely expressed in atherosclerotic lesions affecting native and transplanted human coronary arteries and colocalizes with inducible nitric oxide synthase and nitrotyrosine particularly in macrophages. Arterioscler Thromb Vasc Biol. 1999; 19: 646–655.
20.
Schonbeck U, Sukhova GK, Graber P, Coulter S, Libby P. Augmented expression of cyclooxygenase-2 in human atherosclerotic lesions. Am J Pathol. 1999; 155: 1281–1291.
21.
Belton O, Byrne D, Kearney D, Leahy A, Fitzgerald DJ. Cyclooxygenase-1 and-2-dependent prostacyclin formation in patients with atherosclerosis. Circulation. 2000; 102: 840–845.
22. Matsumoto H, Naraba H, Murakami M, Kudo I, Yamaki K, Ueno A, Oh-ishi S. Concordant induction of prostaglandin E2 synthase with concordant cyclooxygenase-2 leads to preferred production of prostaglandin E2 over thromboxane and prostaglandin D2 in lipopolysaccharide stimulated rat peritoneal macrophages. Biochem Biophys Res Commun. 1997; 230: 110–114.[CrossRef][Medline] [Order article via Infotrieve]
23. Panara MR, Greco A, Santini MG, Rotondo MT, Padovano R, di Giamberardino M, Cipollone F, Cuccurullo F, Patrono C, Patrignani P. Effects of the novel anti-inflammatory compounds NS-398 and L-745,337, on the cyclo-oxygenase activity of human blood prostaglandin endoperoxide synthases. Br J Pharmacol. 1995; 116: 2429–2434.[Medline] [Order article via Infotrieve]
24. Clowes AW, Reidy MA, Clowes MM. Mechanisms of stenosis after arterial injury. Lab Invest. 1998; 49: 208–215.
25.
Morita I, Schindler M, Regier MK, Otto JC, Hori T, DeWitt DL, Smith WL. Different intracellular locations for prostaglandin endoperoxide H synthase-1 and -2. J Biol Chem. 1995; 270: 10902–10908.
26. Harris RC, McKanna JA, Akai Y, Jacobson HR, Dubois RN. Cyclooxygenase-2 is associated with the macula densa of rat kidney and increases with salt restriction. J Clin Invest. 1994; 94: 2504–2510.[Medline] [Order article via Infotrieve]
27. Matsushita M, Masaki Y, Yagi Y, Tanaka T, Wakitani K. Pharmacological profile of JTE-522, a novel prostaglandin H synthase-2 inhibitor, in rats. Inflamm Res. 1997; 46: 461–466.[CrossRef][Medline] [Order article via Infotrieve]
28. Needleman P, Wyche A, Raz A. Platelet and blood vessel arachidonate metabolism and interactions. J Clin Invest. 1979; 63: 345–349.[Medline] [Order article via Infotrieve]
29. Moncada S, Vane JR. Arachidonic acid metabolites and the interactions between platelets and blood vessels. N Engl J Med. 1979; 300: 1142–1147.[Medline] [Order article via Infotrieve]
30. Subbian MTR. Prostaglandin E2 biosynthesis and effect in pigeon aorta. Atherosclerosis. 1978; 29: 487–495.[CrossRef][Medline] [Order article via Infotrieve]
31. Berberian PA, Ziboh VA, Hsia SL. Prostaglandin E2 biosynthesis changes in rabbit aorta and skin during experimental atherogenesis. J Lipid Res. 1976; 17: 46–52.[Abstract]
32. Berberian PA, Ziboh VA, Hsia SL. Inhibition of cholesterol esterification in rabbit aorta by prostaglandin E2. Atherosclerosis. 1977; 27: 213–220.[CrossRef][Medline] [Order article via Infotrieve]
33. Clarke RJ, Mayo G, Price P, Fitzgerald GA. Suppression of thromboxane A2 but not of systemic prostacyclin by controlled-release aspirin. N Engl J Med. 1991; 325: 1137–1141.[Abstract]
34. Coughlin SR, Moskowitz MA, Zetter BR, Antoniades HN, Levine L. Platelet-dependent stimulation of prostacyclin synthesis by platelet derived growth factor. Nature. 1980; 288: 600–602.[CrossRef][Medline] [Order article via Infotrieve]
35. Jones DA, Benjamin CW, Linseman DA. Activation of thromboxane and prostacyclin receptors elicits opposing effects on vascular smooth muscle cell growth and mitogen-activated protein kinase signaling cascades. Mol Pharmacol. 1995; 48: 890–896.[Abstract]
36.
Pakala R, Willerson JT, Benedict CR. Effect of serotonin, thromboxane A2, and specific receptor antagonists on vascular smooth muscle cell proliferation. Circulation. 1997; 96: 2280–2286.
37. Vanhoutte PM. Endothelium-dependent contractions in arteries and veins. Blood Vessels. 1987; 24: 141–144.[Medline] [Order article via Infotrieve]
38.
Kato T, Iwama Y, Okumura K, Hashimoto H, Ito T, Satake T. Prostaglandin H2 may be the endothelium-derived contracting factor released by acetylcholine in the aorta of the rat. Hypertension. 1990; 15: 475–481.
39. Tsujii M, Kawano S, Tsuji S, Sawaoka H, Hori M, DuBois RN. Cyclooxygenase regulates angiogenesis induced by colon cancer cells. Cell. 1998; 93: 705–716.[CrossRef][Medline] [Order article via Infotrieve]
40. Mizuno H, Sakamoto C, Matsuda K, Wada K, Uchida T, Noguchi H, Akamatsu T, Kasuga M. Induction of cyclooxygenase 2 in gastric mucosal lesions and its inhibition by the specific antagonist delays healing in mice. Gastroenterology. 1997; 112: 387–397.[CrossRef][Medline] [Order article via Infotrieve]
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