Donate Help Contact The AHA Sign In Home
American Heart Association
Circulation Research
Search: search_blue_button Advanced Search
Circulation Research. 1996;78:337-342

This Article
Right arrow Abstract Freely available
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow Request Permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Lee, J. S.
Right arrow Articles by Bloch, K. D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Lee, J. S.
Right arrow Articles by Bloch, K. D.
(Circulation Research. 1996;78:337-342.)
© 1996 American Heart Association, Inc.


Articles

Chronic Inhalation of Nitric Oxide Inhibits Neointimal Formation After Balloon-Induced Arterial Injury

Joon S. Lee, Christophe Adrie, Howard J. Jacob, Jesse D. Roberts, Jr, Warren M. Zapol, Kenneth D. Bloch

From the Cardiovascular Research Center and Cardiac Unit of the General Medical Service and Department of Anaesthesia at Massachusetts General Hospital and Harvard Medical School, Boston, Mass.

Correspondence to Kenneth D. Bloch, MD, Cardiovascular Research Center, CNY-4, Massachusetts General Hospital, 149 13th St, Charlestown, MA 02129. E-mail blochk@helix.mgh.harvard.edu.


*    Abstract
up arrowTop
*Abstract
down arrowIntroduction
down arrowMaterials and Methods
down arrowResults
down arrowDiscussion
down arrowReferences
 
Abstract Systemic and local intravascular NO administration inhibits neointimal formation after vascular injury in animal models. NO appears to attenuate smooth muscle proliferation both directly and indirectly by preventing the release of growth factors. Inhalation of low concentrations of NO dilates pulmonary vascular smooth muscle but does not cause systemic vasodilatation. Recently, NO inhalation was found to inhibit platelet function in vivo. We studied the effects of NO inhalation on neointimal formation after balloon-induced injury of the adult rat carotid artery. Beginning 60 minutes before carotid injury, rats breathed either air with 0 or 80 ppm NO for 14 days. Rats were killed, carotid arteries were fixed and paraffin-embedded, and neointimal formation was measured by analyzing the ratio of intimal to medial areas (I/M ratio) in carotid artery cross sections. Intimal hyperplasia was evident in both groups of animals, but I/M ratios were 43% less in animals breathing 80 ppm NO for 2 weeks than in animals breathing air alone (0.78±0.12 and 1.37±0.11 [mean±SE], respectively; P<.02). Similarly, 1 week after carotid injury, neointimal formation was less in rats breathing 80 ppm NO than in rats breathing air alone (I/M ratio, 0.39±0.11 versus 0.76±0.06; P<.02). Breathing 20 ppm NO for 2 weeks or 80 ppm NO for 1 week followed by air alone for 1 week did not attenuate neointimal formation measured at 14 days. In anesthetized rats breathing 80 ppm NO or air alone for 1 hour, neither systemic blood pressure nor bleeding time differed. These observations demonstrate that inhaling 80 ppm NO inhibits neointimal formation after balloon-induced carotid artery injury in rats. NO inhalation may represent a safe and novel method of preventing restenosis after percutaneous angioplasty.


Key Words: nitric oxide • neointimal formation • angioplasty • inhalation therapy


*    Introduction
up arrowTop
up arrowAbstract
*Introduction
down arrowMaterials and Methods
down arrowResults
down arrowDiscussion
down arrowReferences
 
Percutaneous angioplasty is used to treat occlusive arterial disease of both the coronary and the peripheral vascular systems. Unfortunately, although initially successful in >95% of cases, in 30% to 50% of patients, a gradual renarrowing process, which is commonly termed "restenosis," occurs within 6 months.1 This process is characterized by the migration and proliferation of smooth muscle cells in the arterial intima.1 2

Success in attenuating neointimal hyperplasia in animal models by delivering NO to the area of arterial injury has been reported by several investigators. NO delivery was achieved by intravenous administration of an NO donor compound,3 local administration of a gene encoding NO synthase, the enzyme responsible for synthesis of NO from L-arginine,4 and stimulation of endogenous NO production by systemic administration of L-arginine.5 6 7 NO appears to attenuate neointimal formation directly by decreasing smooth muscle cell proliferation8 9 10 and indirectly by inhibiting platelet and leukocyte functions and by stimulating endothelial cell growth.11 12 13

Recently, a novel method of NO delivery was developed: inhalation of low concentrations of NO gas. Inhaled NO readily diffuses into the pulmonary vasculature but, upon reaching the circulation, is rapidly inactivated by hemoglobin.14 15 Inhalation of low concentrations of NO gas selectively dilates the pulmonary vessels without reducing systemic blood pressure, inducing pulmonary injury, or causing methemoglobinemia.16 17 18 19 20 21 NO inhalation does appear to have systemic effects: Hogman and colleagues22 23 reported a 40% prolongation of the bleeding time in rabbits breathing 30 ppm NO. Inhalation of very high concentrations of NO (300 ppm) prolonged the bleeding time 63% but was associated with mild systemic hypotension. Moreover, in a canine model, we recently observed that the inhalation of 20 and 80 ppm NO decreased platelet-mediated coronary artery reocclusion after thrombolysis without causing systemic hypotension or methemoglobinemia.24

In the present study, the ability of inhaled NO to attenuate neointimal formation after balloon-induced carotid artery injury was studied in rats. Continuous inhalation of 80 ppm NO significantly decreased the neointimal formation in response to vascular injury.


*    Materials and Methods
up arrowTop
up arrowAbstract
up arrowIntroduction
*Materials and Methods
down arrowResults
down arrowDiscussion
down arrowReferences
 
Arterial Injury
Adult male Sprague-Dawley rats (Charles River Laboratories, Wilmington, Mass) underwent balloon-induced injury of the common carotid artery using methods previously described to induce consistent neointimal hyperplasia.25 Rats weighing 300 to 350 g were anesthetized by intraperitoneal injection of ketamine (60 to 80 mg/kg) and acepromazine (0.1 mg/kg). A longitudinal midline cervical incision was made, and the left carotid artery was isolated. After dissection of the internal and external carotid bifurcation, the distal external carotid segment was ligated with a 4-0 silk suture. A small arteriotomy was made in the external carotid, and a 2F Fogarty balloon catheter (Baxter Edwards LIS) was inserted through the arteriotomy and advanced {approx}2 cm below the carotid bifurcation. The balloon was distended with saline to 2 atm using an ACS Inflator Plus (Advanced Cardiovascular Systems Inc) and was gently withdrawn to the level of the bifurcation. The balloon was deflated, and the balloon-induced injury was repeated for a total of three times. The balloon catheter was then withdrawn, and after allowing backbleeding through the arteriotomy site to eliminate any potential thrombus or air bubbles, the external carotid was ligated proximal to the arteriotomy site using 4-0 silk sutures. After visual inspection to ensure adequate pulsation of the common carotid artery, the surgical incision was closed, and the rats were allowed to recover from anesthesia.

Chronic NO Inhalation
Rats breathed NO in specially prepared 40-L acrylic inhalation chambers.26 The gas mixtures were blended using separately regulated and calibrated flowmeters for oxygen, pressurized air, and NO stock gases (800 and 10 000 ppm, Airco). The oxygen concentration in the effluent gas from the chamber was analyzed periodically throughout the experimental period to ensure an FIO2 of 0.21 using a polarographic electrode (Oxygen Monitor, Hudson Ventronics Division). Concentrations of NO, as well as nitrogen dioxide and nitrogen with higher oxidation states (NOx), in the effluent gas were measured with an NO/NOx chemiluminescence analyzer27 (model 14A, Thermo Environmental Instruments Inc). Soda lime (J.T. Baker Chemical Co), which was maintained in the chambers to reduce the levels of NOx, was replaced twice a week. NO concentration was maintained at 20 or 80 ppm, depending on the experiment. Serial measurements revealed that NOx levels were <=6% of the NO concentration. The gases exiting the exposure chambers, as well as those discharging from the chemiluminescence instrument, were scavenged with a Venturi trap maintained at negative atmospheric pressure by the laboratory's central vacuum system.

Control animals were maintained in filtered cages in the same room as the NO-treated animals. NO exposure was begun {approx}60 minutes before surgery. During the surgical procedure, the NO-treated group was taken out of the chamber and exposed to NO by a modified face mask fed from the inflow gas tubing of the chambers. Analysis of the gas in the face mask revealed NO levels of 50 to 80 ppm during the 80-ppm experiments and 10 to 20 ppm during the 20-ppm studies. The total duration of time out of the chamber and under the face mask ranged from 20 to 40 minutes.

Rats were given free access to water and rat chow throughout the study. Fifteen rats breathing air alone, 8 rats breathing 20 ppm NO, and 7 rats breathing 80 ppm NO were weighed before carotid injury and at the time of death 14 days later.

Morphometric Analysis
Rats were euthanized by an intraperitoneal injection of sodium pentobarbital. A 16-gauge catheter was introduced into the ascending aorta via the left ventricular apex. The descending thoracic aorta was ligated, and the right atrium was vented. The vasculature was then perfused at a pressure of 100 mm Hg with 100 mL of normal saline, followed by 50 mL of 2% paraformaldehyde in PBS. After in vivo fixation for 15 to 30 minutes, both common carotid arteries were excised and underwent additional fixation overnight in PBS containing 4% paraformaldehyde. The tissue was dehydrated using sequentially increasing concentrations of ethanol followed by xylene and embedded in paraffin. Cross sections (6 µm) were cut and stained with hematoxylin and eosin and/or elastin stain for analysis. For each animal, a single section 7 to 8 mm proximal to the carotid bifurcation was photographed, and the image was digitized at 2000 dots per inch using a Kodak RF 2035 Plus Film Scanner (Eastman Kodak Co). The carotid artery intimal and medial areas were calculated using an image analysis computer program (NIH Image). Section preparation, photography, and morphometric analysis were performed by an investigator blinded to the experimental group.

Bleeding Time and Blood Pressure Measurements
Twenty rats weighing 350 to 450 g were anesthetized as described above. For each rat, the right common carotid artery was cannulated with a polyethylene catheter (model PE-50, Intramedic, Becton Dickinson), and a tracheostomy was performed. After paralysis with 0.2 mg/kg pancuronium (Gensia Pharmaceuticals, Inc), each animal was ventilated at 50 breaths per minute and a tidal volume of 5 mL/kg with a rodent ventilator (model 683, Harvard Apparatus). FIO2 was kept constant at 0.21. Carotid arterial blood pressure was continuously measured with calibrated pressure transducers (Cobe Laboratories) and recorded (model 7754, Hewlett Packard). Bleeding times were performed at room temperature with the tip of the tail 10 cm below the platform upon which the animal was placed. Bleeding was induced by transection of the tail 5 mm from the tip. The tails were gently blotted with filter paper, and the time in seconds to cessation of bleeding was noted by an investigator blinded to the inhaled NO concentration.

After ventilation with air for a 20-minute stabilization period, rats were ventilated with either 0 (n=10) or 80 (n=10) ppm NO for 60 minutes. Bleeding times were performed at the end of this exposure period. To ensure consistent oxygenation and ventilation, heparinized blood samples were obtained from the carotid artery during the stabilization period and during the 60-minute exposure period, and blood gas tension and pH were measured on a blood gas analyzer (model 238, Ciba Corning Diagnostics Ltd).

Statistical Analysis
All data are presented as mean±SE. Intimal and medial areas and ratios of intimal to medial areas (I/M ratios) of the four groups of rats studied 14 days after carotid injury were compared by a factorial model of ANOVA. When significant differences were detected, Scheffé's analysis was used post hoc to compare groups. Intimal and medial areas and I/M ratios of the two groups of rats studied 7 days after carotid injury were compared using an unpaired Student's t test. Significance was declared at P<.05.


*    Results
up arrowTop
up arrowAbstract
up arrowIntroduction
up arrowMaterials and Methods
*Results
down arrowDiscussion
down arrowReferences
 
A total of 82 rats underwent the carotid artery injury procedure. There were no postprocedure deaths. All rats, regardless of their experimental group, exhibited normal grooming behavior and activity levels throughout the study. Other than occasional ptosis on the side of the carotid injury, no rats exhibited a gross neurological deficit. No significant differences were observed in weight gained over the 2-week experimental period in rats breathing 0, 20, or 80 ppm NO (data not shown).

In the first series of experiments, the rats undergoing balloon-induced injury to the carotid artery breathed either air (n=35) or 80 ppm NO in air (n=20) throughout the duration of the study. The animals were killed 14 days after injury for morphometric analysis of the injured carotid artery. Upon light microscopic examination, three rats breathing air alone and one breathing NO were found to have thrombotic occlusion of the lumen of the injured carotid artery. These four animals were excluded from further analysis. Vessels harvested from both groups exhibited loss of endothelium and the development of neointimal hyperplasia as described previously by other investigators.25 Examination by light microscopy did not reveal a qualitative difference in the cellular morphology of the neointima between animals breathing NO and those that did not (FigureDown). Quantitative analysis revealed that the intimal area was 45% less in rats breathing 80 ppm NO than in rats breathing air alone (P<.04), whereas medial areas did not differ between the two groups (TableDown). The I/M ratio was 43% less in animals inhaling 80 ppm NO for 14 days than in animals breathing air (P<.02).



View larger version (0K):
[in this window]
[in a new window]
 
Figure 1. Cross sections of balloon-injured rat carotid arteries (original magnification x100). Sections of carotid arteries are shown 14 days after injury from rats exposed to air alone (top) and exposed to air with 80 ppm NO (bottom). The sections shown represent carotid arteries with the median intimal-to-medial ratios for the two groups.


View this table:
[in this window]
[in a new window]
 
Table 1. Morphometric Analysis of Injured Rat Carotid Arteries

The effect of breathing NO on neointimal formation was also examined 1 week after carotid injury. Neointimal formation was less in carotid arteries 1 week after injury than 2 weeks after injury. One week after carotid injury, inhaling 80 ppm NO decreased neointimal formation by 55% (P<.01) but did not alter the medial area. The I/M ratio in injured carotid arteries from rats breathing 80 ppm was 49% less than that in arteries from rats breathing air (P<.02).

Two additional groups of animals were studied. To investigate the effect of breathing a lower concentration of NO on the neointimal response, 8 rats subjected to carotid injury were exposed to 20 ppm NO for 2 weeks and then killed. In these rats, the carotid artery I/M ratio was 0.92±0.23 and did not differ from that observed in rats breathing air alone. To determine whether shorter exposures to NO were sufficient to attenuate neointimal formation, 7 rats subjected to carotid artery injury breathed air with 80 ppm NO for 1 week, followed by breathing air alone for a second week. In these rats, the I/M ratio was 1.06±0.24 and was not different from that observed in rats breathing air for 2 weeks.

To investigate the effect of NO inhalation on systemic platelet function, tail transection bleeding times were measured in anesthetized rats breathing NO for 1 hour. Bleeding time did not differ between 10 rats breathing air (213±20 seconds) and 10 rats breathing 80 ppm NO (246±18 seconds). Systemic blood pressures were also measured in these animals. Mean arterial blood pressures did not differ significantly in rats breathing air (69±3 mm Hg) or 80 ppm NO (78±5 mm Hg).


*    Discussion
up arrowTop
up arrowAbstract
up arrowIntroduction
up arrowMaterials and Methods
up arrowResults
*Discussion
down arrowReferences
 
In the present study, chronic inhalation of low concentrations of NO gas was found to inhibit neointimal formation after balloon-induced vascular injury. Breathing 80 ppm NO decreased neointimal formation 1 and 2 weeks after injury (TableUp). To achieve this inhibition, the rats needed to continuously breathe NO: inhaling NO for the first 7 days after injury followed by 7 days of air breathing did not inhibit neointimal formation at 14 days. Breathing a lower concentration of inhaled NO (20 ppm) for 2 weeks did not attenuate the neointimal response to injury.

Ventilation of anesthetized rats with 80 ppm NO did not alter systemic blood pressure. Although exposures of unanesthetized rats might have provided different results, studies in awake sheep revealed that breathing 80 ppm NO did not alter systemic blood pressure.16

Although inhibition of neointimal formation by local or systemic delivery of NO donor compounds has been observed previously,3 5 6 7 28 this is the first report of producing such an effect using the novel respiratory therapy, NO inhalation. The magnitude of inhibition achieved by inhaling 80 ppm NO was comparable to that reported for the administration of L-arginine.5 6 7 Our findings were also similar to those of Guo et al,3 who observed that intravenous administration of the NO donor compound SPM-5185 decreased neointimal formation in rat carotid arteries 7 days after desiccation-induced intimal injury and did not cause systemic hypotension.

Several mechanisms potentially account for the inhibition of neointimal formation by NO inhalation. Many investigators have postulated that the effect of NO on neointimal hyperplasia is mediated by its ability to directly inhibit smooth muscle cell proliferation at the site of injury. This is supported by in vitro experiments showing that NO donor compounds suppress the proliferation of isolated vascular smooth muscle cells3 8 9 29 ; however, recent evidence suggests that under certain conditions NO may also stimulate smooth muscle cell mitogenesis.30

Because NO is almost immediately inactivated in the presence of hemoglobin, NO has an extremely short half-life in blood of 111 to 130 milliseconds.29 Using chemiluminescence, Rich et al15 measured the concentration of NO in the effluent of isolated perfused rat lungs ventilated with 1000 ppm NO; they observed that if the lungs were perfused with whole blood, NO was undetectable in the effluent. The rapid inactivation of NO by hemoglobin accounts for the ability of inhaled NO to produce local pulmonary vasodilatation without reducing the systemic vascular resistance or systemic arterial pressure in animal models and in humans.16 17 18 19 20 21 Therefore, it is unlikely that NO absorbed into the pulmonary circulation decreases neointimal formation at the site of carotid injury via a direct effect.

Under certain conditions, NO can react with thiol-containing molecules, which then have NO-like vasodilatory and platelet inhibitory properties.31 NO bound to thiols has a longer functional half-life in blood than does free NO. During NO inhalation, S-nitrosothiols may be produced in the pulmonary vasculature or formed in the airways32 and then absorbed into the circulation. These S-nitrosothiols could travel to the site of arterial injury and produce the same effect on neointimal formation as NO donor compounds. Rimar and Gillis14 measured the vasodilator activity present in the effluent of isolated perfused rabbit lungs ventilated with 150 ppm NO; they observed that even small amounts of blood in the perfusate blocked the ability of the effluent to vasodilate aortic rings. These results suggest that circulating S-nitrosothiols formed during NO inhalation possess minimal vasodilator properties or are present in very low concentrations and are unlikely to directly inhibit neointimal formation.

Another mechanism potentially accounting for the effect of inhaled NO on neointimal formation may be that exposure of one or more circulating blood elements to NO, during transit through the pulmonary circulation, modulates their function at the site of vascular injury, inhibiting neointimal development. Through this mechanism, inhaled NO would be able to exert a systemic vascular effect despite being anatomically limited to the pulmonary vasculature. Although the design of the present study does not directly reveal which circulating blood element mediates the effect of inhaled NO, candidates include platelets and leukocytes.

Many investigators have demonstrated the platelet-inhibitory effects of NO and NO donor compounds.13 33 34 In vitro, NO inhibits platelet adhesion,35 36 activation,37 aggregation,35 38 and mitogen release.39 In vivo, systemic administration of NO donor compounds to animals and humans prolongs the bleeding time40 41 and inhibits platelet adherence to atherosclerotic plaques,42 as well as to balloon-injured arteries.3 43 44 The importance of platelets for neointimal formation after vascular injury is well established. Friedman et al45 demonstrated prevention of neointimal formation after balloon-induced injury in rabbits by inducing thrombocytopenia using antiplatelet antisera. In the injured rat carotid artery model, Fingerle et al46 observed that induction of thrombocytopenia for 48 to 96 hours attenuated neointimal formation measured 4 and 7 days after injury.

In contrast to the observations of Hogman and colleagues,22 23 who reported a modest prolongation of the bleeding time in humans and animals during NO inhalation, we did not observe a prolongation of the tail-transection bleeding time in rats breathing 80 ppm NO for 1 hour. The tail-transection bleeding times that we measured, which reflect both platelet and coagulation function, were similar to those reported by de Gaetano and colleagues.47 48 It is possible that shorter or longer durations of exposure to NO might have prolonged the bleeding time. The failure of inhaled NO to prolong the bleeding time does not exclude the possibility that inhaled NO attenuated neointimal formation by altering systemic platelet function. Samama et al49 recently reported that inhalation of up to 100 ppm NO by six patients with adult respiratory distress syndrome did not prolong the bleeding time but did decrease ex vivo platelet aggregation. We observed that inhalation of 20 and 80 ppm NO increased vascular patency in a canine model of platelet-mediated coronary artery thrombosis after thrombolysis24 but did not prolong the bleeding time measured on the ventral aspect of the tongue and on the shaved forepaw (authors' unpublished data, 1995).

Another potential cellular mediator of the effect of inhaled NO on neointimal formation is the leukocyte. It is accepted that leukocytes, especially monocytes, play an important role in the pathogenesis of primary atherosclerosis as well as restenosis.50 In vitro, NO inhibits many leukocyte functions, including adhesion51 and chemotaxis.12 It remains to be determined whether NO inhalation alters systemic leukocyte function or whether NO-exposed leukocytes modulate neointimal formation.

Based on these observations from a single rodent model of vascular injury, it cannot be concluded that inhaled NO will prevent restenosis after percutaneous coronary angioplasty in humans. However, several features of inhaled NO therapy suggest that it may be clinically useful. First, NO gas may be readily administered via face mask or nasal prongs. Second, no significant side effects have been reported associated with inhaling NO in the low concentrations used in clinical studies (up to 80 ppm). Third, NO inhalation in humans, as well as rats, does not cause systemic hypotension. Fourth, breathing NO causes little or no prolongation of the bleeding time in the four species studied (human, dog, rabbit, and rat), suggesting that its clinical use will be associated with a low risk of hemorrhagic complications. A potential disadvantage of inhaled NO for the prevention of neointimal formation is that continuous inhalation appears to be necessary: neointimal formation in animals breathing 80 ppm NO for 1 week followed by breathing air for 1 week did not differ from that in animals breathing air alone for 2 weeks. This latter observation is similar to the finding that transient thrombocytopenia decreased neointimal formation when measured 1 week after injury but not when measured 2 weeks after injury.46 It is conceivable that agents, such as selective inhibitors of type V cGMP phosphodiesterase,52 will be used to augment and/or prolong the effect of inhaled NO or to permit intermittent administration. Alternatively, inhaled NO may be used to postpone neointimal formation, during which time other agents may be used, for example, to accelerate reendothelialization of injured vascular segments.53

In summary, inhalation of 80 ppm NO inhibits the neointimal response to balloon-induced injury of the rat carotid artery. NO inhalation did not cause systemic hypotension or prolong the bleeding time. These results suggest that inhaled NO may provide a safe respiratory therapy to inhibit clinical restenosis.


*    Acknowledgments
 
This study was supported by National Institutes of Health grants T32 HL-07208 (Dr Lee), HL-42397 (Drs Zapol and Roberts), and HL-45895 (Dr Bloch) and by a grant to the Cardiovascular Research Center from Bristol Myers Squibb Pharmaceuticals. We would like to thank Chris Simpson for his expert assistance and advice in the preparation of the histological sections and Jonathan Delgado for his help with the computer software.

Received September 5, 1995; accepted November 22, 1995.


*    References
up arrowTop
up arrowAbstract
up arrowIntroduction
up arrowMaterials and Methods
up arrowResults
up arrowDiscussion
*References
 

  1. Liu MW, Roubin GS, King SB. Restenosis after coronary angioplasty: potential biologic determinants and role of intimal hyperplasia. Circulation. 1989;79:1374-1387. [Abstract/Free Full Text]
  2. Austin GE, Ratliff NB, Hollman J, Tabei S, Phillips DF. Intimal proliferation of smooth muscle as an explanation for recurrent coronary artery restenosis after percutaneous transluminal coronary angioplasty. J Am Coll Cardiol. 1985;6:369-375. [Abstract]
  3. Guo J, Milhoan KA, Tuan RS, Lefer AM. Beneficial effect of SPM-5185, a cysteine-containing nitric oxide donor, in rat carotid artery intimal injury. Circ Res. 1994;75:77-84. [Abstract/Free Full Text]
  4. van der Leyen HE, Gibbons GH, Morishita R, Lewis NP, Zhang L, Nakajima M, Kaneda Y, Cooke JP, Dzau V. Gene therapy inhibiting neointimal vascular lesion: in vivo transfer of endothelial cell nitric oxide synthase gene. Proc Natl Acad Sci U S A. 1995;92:1137-1141. [Abstract/Free Full Text]
  5. Tarry WC, Makhoul RG. L-Arginine improves endothelium-dependent vasorelaxation and reduces intimal hyperplasia after balloon angioplasty. Arterioscler Thromb. 1994;14:938-943. [Abstract/Free Full Text]
  6. Hamon M, Vallet B, Bauters C, Wernert N, McFadden EP, Lablanche J-M, Dupuis B, Bertrand M. Long-term oral administration of L-arginine reduces intimal thickening and enhances neoendothelium-dependent acetylcholine-induced relaxation after arterial injury. Circulation. 1994;90:1357-1362. [Abstract/Free Full Text]
  7. McNamara DB, Bedi B, Aurora H, Tena L, Ignarro LJ, Kadowitz PJ, Akers DL. L-Arginine inhibits balloon catheter-induced intimal hyperplasia. Biochem Biophys Res Commun. 1993;193:291-296. [Medline] [Order article via Infotrieve]
  8. Assender JW, Southgate KM, Newby AC. Does nitric oxide inhibit smooth muscle proliferation? J Cardiovasc Pharmacol. 1991;17:S104-S107.
  9. Garg UC, Hassid A. Nitric oxide-generating vasodilators and 8-bromocyclic guanosine monophosphate inhibit mitogenesis and proliferation of cultured rat smooth muscle cells. J Clin Invest. 1989;83:1774-1777.
  10. Kariya K, Kawahara Y, Araki S, Fukuzaki H, Takai Y. Antiproliferative action of cyclic GMP-elevating vasodilators in cultured rabbit aortic smooth muscle cells. Atherosclerosis. 1989;80:143-147. [Medline] [Order article via Infotrieve]
  11. Ziche M, Morbidelli L, Masini E, Granger H, Geppetti P, Ledda F. Nitric oxide promotes DNA synthesis and cyclic GMP formation in endothelial cells from postcapillary venules. Biochem Biophys Res Commun. 1993;192:1198-1203. [Medline] [Order article via Infotrieve]
  12. Bath PM. The effect of nitric oxide-donating vasodilators on monocyte chemotaxis and intracellular cGMP concentrations in vitro. Eur J Clin Pharmacol. 1993;45:53-58. [Medline] [Order article via Infotrieve]
  13. Bassenge E. Antiplatelet effects of endothelium-derived relaxing factor and nitric oxide donors. Eur Heart J. 1991;12:S12-S15.
  14. Rimar S, Gillis CN. Selective pulmonary vasodilation by inhaled nitric oxide is due to hemoglobin inactivation. Circulation. 1993;88:2884-2887. [Abstract/Free Full Text]
  15. Rich CF, Roos CM, Anderson SM, Urich DC, Daugherty MO, Johns RA. Inhaled nitric oxide: dose response and the effects of blood in the isolated rat lung. J Appl Physiol. 1993;75:1278-1284. [Abstract/Free Full Text]
  16. Frostell C, Fratacci MD, Wain JC, Jones R, Zapol WM. Inhaled nitric oxide: a selective pulmonary vasodilator reversing hypoxic pulmonary vasoconstriction. Circulation. 1991;83:2038-2047. [Abstract/Free Full Text]
  17. Kouyoumdijian C, Adnot S, Levame N, Eddahibi S, Bousbaa H. Continuous inhalation of nitric oxide protects against development of pulmonary hypertension in chronically hypoxic rats. J Clin Invest. 1994;94:578-584.
  18. Roberts JD, Lang P, Bigatello LM, Vlahakes GJ, Zapol WM. Inhaled nitric oxide in congenital heart disease. Circulation. 1993;87:447-453. [Abstract/Free Full Text]
  19. Roberts JD Jr, Chen T-Y, Kawai N, Wain J, Dupuy P, Shimouchi A, Bloch KD, Polaner D, Zapol WM. Inhaled nitric oxide reverses pulmonary vasoconstriction in the hypoxic and acidotic newborn lamb. Circ Res. 1993;72:246-254. [Abstract/Free Full Text]
  20. Pepke-Zaba J, Higenbottam TW, Dinh-Xuan AT, Stone D, Wallwork J. Inhaled nitric oxide as a cause of selective pulmonary vasodilation in pulmonary hypertension. Lancet. 1991;338:1173-1174. [Medline] [Order article via Infotrieve]
  21. Semigran MJ, Cockrill BA, Kacmarek R, Thompson BT, Zapol WM, Dec GW, Fifer MA. Hemodynamic effects of inhaled nitric oxide in heart failure. J Am Coll Cardiol. 1994;24:982-988. [Abstract]
  22. Hogman M, Frostell C, Arnberg H, Hedenstierna G. Bleeding time prolongation and NO inhalation. Lancet. 1993;341:1664-1665. [Medline] [Order article via Infotrieve]
  23. Hogman M, Frostell C, Arnberg H, Sandhagen B, Hedenstierna G. Prolonged bleeding time during nitric oxide inhalation in the rabbit. Acta Physiol Scand. 1994;151:125-129. [Medline] [Order article via Infotrieve]
  24. Adrie C, Moreno PR, Bloch KD, Hurford WE, Guerrero L, Holt R, Zapol WM, Gold HK, Semigran MJ. Inhaled nitric oxide increases coronary artery patency after thrombolysis. FASEB J. 1995;9:A876. Abstract.
  25. Clowes AW, Reidy MA, Clowes MM. Kinetics of cellular proliferation after arterial injury, I: smooth muscle growth in the absence of endothelium. Lab Invest. 1983;49:327-333. [Medline] [Order article via Infotrieve]
  26. Roberts JD Jr, Roberts CT, Jones RC, Zapol WM, Bloch KD. Continuous nitric oxide inhalation reduces pulmonary arterial structural changes, right ventricular hypertrophy, and growth retardation in the hypoxic newborn rat. Circ Res. 1995;76:215-222. [Abstract/Free Full Text]
  27. Fontijin A, Sabadell AJ, Ronco RJ. Homogenous chemiluminescent measurement on nitric oxide with ozone. Anal Chem. 1970;42:575-579.
  28. Taguchi J, Abe J, Okazaki H, Takuwa Y, Kurokawa K. L-Arginine inhibits neointimal formation following balloon injury. Life Sci. 1993;53:387-392.
  29. Kelm M, Schrader J. Control of coronary vascular tone by nitric oxide. Circ Res. 1990;66:1561-1575. [Abstract/Free Full Text]
  30. Hassid A, Arabshahi H, Boucier T, Dhaunsi GS, Matthews C. Nitric oxide selectively amplifies FGF-2-induced mitogenesis in primary rat aortic smooth muscle cells. Am J Physiol. 1994;267:H1040-H1048. [Abstract/Free Full Text]
  31. Stamler JS, Simon DI, Osborne JA, Mullins ME, Jaraki O, Michel T, Singel DJ, Loscalzo J. S-Nitrosylation of proteins with nitric oxide: synthesis and characterization of biologically active compounds. Proc Natl Acad Sci U S A. 1992;89:444-448. [Abstract/Free Full Text]
  32. Gaston B, Reilly J, Drazen JM, Fackler J, Ramdev P, Arnelle D, Mullins ME, Sugarbaker DJ, Chee C, Singel DJ, Loscalzo J, Stamler JS. Endogenous nitrogen oxides and bronchodilator S-nitrosothiols in human airways. Proc Natl Acad Sci U S A. 1993;90:10957-10961. [Abstract/Free Full Text]
  33. Folts JD, Stamler J, Loscalzo J. Intravenous nitroglycerin infusion inhibits cyclic blood flow responses caused by periodic platelet thrombus formation in stenosed canine coronary arteries. Circulation. 1991;83:2122-2127. [Abstract/Free Full Text]
  34. Loscalzo J. Antiplatelet and antithrombotic effects of organic nitrates. Am J Cardiol. 1992;70:18B-22B. [Medline] [Order article via Infotrieve]
  35. Radomski MW, Palmer RMJ, Moncada S. The role of nitric oxide and cGMP in platelet adhesion to vascular endothelium. Biochem Biophys Res Commun. 1987;148:1482-1489. [Medline] [Order article via Infotrieve]
  36. deGraaf JC, Banga JD, Moncada S, Palmer RMJ, deGroot PG, Sixma JJ. Nitric oxide functions as an inhibitor of platelet adhesion under flow conditions. Circulation. 1992;85:2284-2290. [Abstract/Free Full Text]
  37. Radomski MW, Rees DD, Dutra A, Moncada S. S-Nitroso-glutathione inhibits platelet activation in vitro and in vivo. Br J Pharmacol. 1992;107:745-749. [Medline] [Order article via Infotrieve]
  38. Furlong B, Henderson AH, Lewis MJ, Smith JA. Endothelium-derived relaxing factor inhibits in vitro platelet aggregation. Br J Pharmacol. 1987;90:687-692. [Medline] [Order article via Infotrieve]
  39. Barrett ML, Willis AL, Vane JR. Inhibition of platelet-derived mitogen release by ntric oxide. Agents Actions. 1989;27:488-491. [Medline] [Order article via Infotrieve]
  40. Ring T, Knudsen F, Kristensen SD, Larsen CE. Nitroglycerin prolongs the bleeding time in healthy males. Thromb Res. 1983;29:553-559. [Medline] [Order article via Infotrieve]
  41. Simon DI, Stamler JS, Jaraki O, Keaney JF, Osborne JA, Francis SA, Singel DJ, Loscalzo J. Antiplatelet properties of protein S-nitrothiols derived from nitric oxide and endothelium-derived relaxing factor. Arterioscler Thromb. 1993;13:791-799. [Abstract/Free Full Text]
  42. Sinzinger H, Fitscha P, O'Grady J, Rauscha F, Rogatti W, Vane JR. Synergistic effect of prostaglandin E1 and isosorbide dinitrate in peripheral vascular disease. Lancet. 1990;335:627-628. [Medline] [Order article via Infotrieve]
  43. Groves PH, Penny WJ, Cheadle HA, Lewis MJ. Exogenous nitric oxide inhibits in vivo platelet adhesion following balloon angioplasty. Cardiovasc Res. 1992;26:615-619. [Abstract/Free Full Text]
  44. Groves PH, Lewis MJ, Cheadle HA, Penny WJ. SIN-1 reduces platelet adhesion and platelet thrombus formation in a porcine model of balloon angioplasty. Circulation. 1993;87:590-597. [Abstract/Free Full Text]
  45. Friedman RJ, Stemerman MB, Wenz B, Moore S, Gauldie J, Gent M, Tiell ML, Spaet TH. The effect of thrombocytopenia on experimental arteriosclerotic lesion formation in rabbits. J Clin Invest. 1977;60:1191-1201.
  46. Fingerle J, Johnson R, Clowes AW, Majesky MW, Reidy MA. Role of platelets in smooth muscle cell proliferation and migration after vascular injury in rat carotid artery. Proc Natl Acad Sci U S A. 1989;86:8412-8416. [Abstract/Free Full Text]
  47. Dejana E, Villa S, deGaetano G. Bleeding time in rats: a comparison of different experimental conditions. Thromb Haemost. 1982;48:108-111. [Medline] [Order article via Infotrieve]
  48. Buczko W, Gambino MC, deGaetano G. Prolongation of rat tail bleeding time by ketanserin: mechanism of action. Eur J Pharmacol. 1984;103:261-268. [Medline] [Order article via Infotrieve]
  49. Samama CM, Diaby M, Fellahi J-L, Mdahafa A, Eyraud D, Arock M, Guillosson J-J, Coriat P, Rouby J-J. Inhibition of platelet aggregation by inhaled nitric oxide in patients with acute respiratory distress syndrome. Anesthesiology. 1995;83:56-65. [Medline] [Order article via Infotrieve]
  50. Ross R. The pathogenesis of atherosclerosis a perspective for the 1990s. Nature. 1993;362:801-809. [Medline] [Order article via Infotrieve]
  51. Kubes P, Suzuki M, Granger DN. Nitric oxide: an endogenous modulator of leukocyte adhesion. Proc Natl Acad Sci U S A. 1991;88:4651-4655. [Abstract/Free Full Text]
  52. Ichinose F, Adrie C, Hurford WE, Zapol WM. Prolonged pulmonary vasodilator action of inhaled nitric oxide by zaprinast in awake lambs. J Appl Physiol. 1995;78:1288-1295. [Abstract/Free Full Text]
  53. Asahara T, Bauters C, Pastore C, Kearney M, Rossow S, Bunting S, Ferrara N, Symes JF, Isner JM. Local delivery of vascular endothelial growth factor accelerates reendothelialization and attenuates intimal hyperplasia in balloon-injured rat carotid artery. Circulation. 1995;91:2793-2801.[Abstract/Free Full Text]



This article has been cited by other articles:


Home page
J Am Coll CardiolHome page
X. Liu, Y. Huang, P. Pokreisz, P. Vermeersch, G. Marsboom, M. Swinnen, E. Verbeken, J. Santos, M. Pellens, H. Gillijns, et al.
Nitric Oxide Inhalation Improves Microvascular Flow and Decreases Infarction Size After Myocardial Ischemia and Reperfusion
J. Am. Coll. Cardiol., August 21, 2007; 50(8): 808 - 817.
[Abstract] [Full Text] [PDF]


Home page
Am. J. Physiol. Heart Circ. Physiol.Home page
R. Hataishi, A. C. Rodrigues, T. G. Neilan, J. G. Morgan, E. Buys, S. Shiva, R. Tambouret, D. S. Jassal, M. J. Raher, E. Furutani, et al.
Inhaled nitric oxide decreases infarction size and improves left ventricular function in a murine model of myocardial ischemia-reperfusion injury
Am J Physiol Heart Circ Physiol, July 1, 2006; 291(1): H379 - H384.
[Abstract] [Full Text] [PDF]


Home page
Am. J. Physiol. Heart Circ. Physiol.Home page
R. Hataishi, W. M. Zapol, K. D. Bloch, and F. Ichinose
Inhaled nitric oxide does not reduce systemic vascular resistance in mice
Am J Physiol Heart Circ Physiol, May 1, 2006; 290(5): H1826 - H1829.
[Abstract] [Full Text] [PDF]


Home page
J. Lipid Res.Home page
Z. W. Q. Moore and D. Y. Hui
Apolipoprotein E inhibition of vascular hyperplasia and neointima formation requires inducible nitric oxide synthase
J. Lipid Res., October 1, 2005; 46(10): 2083 - 2090.
[Abstract] [Full Text] [PDF]


Home page
Am. J. Physiol. Heart Circ. Physiol.Home page
D. Zhuang, A.-C. Ceacareanu, B. Ceacareanu, and A. Hassid
Essential role of protein kinase G and decreased cytoplasmic Ca2+ levels in NO-induced inhibition of rat aortic smooth muscle cell motility
Am J Physiol Heart Circ Physiol, April 1, 2005; 288(4): H1859 - H1866.
[Abstract] [Full Text] [PDF]


Home page
HypertensionHome page
J. P. Dina, T. Feres, A. C.M. Paiva, and T. B. Paiva
Role of Membrane Potential and Expression of Endothelial Factors in Restenosis After Angioplasty in SHR
Hypertension, January 1, 2004; 43(1): 131 - 135.
[Abstract] [Full Text] [PDF]


Home page
Arterioscler. Thromb. Vasc. Bio.Home page
P. Maffia, A. Ianaro, R. Sorrentino, L. Lippolis, F. M. Maiello, P. del Soldato, A. Ialenti, and G. Cirino
Beneficial Effects of NO-Releasing Derivative of Flurbiprofen (HCT-1026) in Rat Model of Vascular Injury and Restenosis
Arterioscler. Thromb. Vasc. Biol., February 1, 2002; 22(2): 263 - 267.
[Abstract] [Full Text] [PDF]


Home page
Proc. Natl. Acad. Sci. USAHome page
C. Napoli, G. Aldini, J. L. Wallace, F. de Nigris, R. Maffei, P. Abete, D. Bonaduce, G. Condorelli, F. Rengo, V. Sica, et al.
Efficacy and age-related effects of nitric oxide-releasing aspirin on experimental restenosis
PNAS, January 24, 2002; (2002) 22639399.
[Abstract] [Full Text] [PDF]


Home page
Am. J. Physiol. Lung Cell. Mol. Physiol.Home page
A. N. MacRitchie, K. H. Albertine, J. Sun, P. S. Lei, S. C. Jensen, A. A. Freestone, P. M. Clair, M. J. Dahl, E. A. Godfrey, D. P. Carlton, et al.
Reduced endothelial nitric oxide synthase in lungs of chronically ventilated preterm lambs
Am J Physiol Lung Cell Mol Physiol, October 1, 2001; 281(4): L1011 - L1020.
[Abstract] [Full Text] [PDF]


Home page
Am. J. Physiol. Heart Circ. Physiol.Home page
C. Brown, Y. Lin, and A. Hassid
Requirement of protein tyrosine phosphatase SHP2 for NO-stimulated vascular smooth muscle cell motility
Am J Physiol Heart Circ Physiol, October 1, 2001; 281(4): H1598 - H1605.
[Abstract] [Full Text] [PDF]


Home page
J Am Coll CardiolHome page
S. Kaul, B. Cercek, J. Rengstrom, X.-P. Xu, M. D. Molloy, P. Dimayuga, A. K. Parikh, M. C. Fishbein, J. Nilsson, T. B. Rajavashisth, et al.
Polymeric-based perivascular delivery of a nitric oxide donor inhibits intimal thickening after balloon denudation arterial injury: role of nuclear factor-kappaB
J. Am. Coll. Cardiol., February 1, 2000; 35(2): 493 - 501.
[Abstract] [Full Text] [PDF]


Home page
Circ. Res.Home page
K.-Y. Chyu, P. Dimayuga, J. Zhu, J. Nilsson, S. Kaul, P. K. Shah, and B. Cercek
Decreased Neointimal Thickening After Arterial Wall Injury in Inducible Nitric Oxide Synthase Knockout Mice
Circ. Res., December 3, 1999; 85(12): 1192 - 1198.
[Abstract] [Full Text] [PDF]


Home page
Cardiovasc ResHome page
C. S Hayward, R. P Kelly, and P. S Macdonald
Inhaled nitric oxide in cardiology practice
Cardiovasc Res, August 15, 1999; 43(3): 628 - 638.
[Abstract] [Full Text] [PDF]


Home page
Cardiovasc ResHome page
M. Kibbe, T. Billiar, and E. Tzeng
Inducible nitric oxide synthase and vascular injury
Cardiovasc Res, August 15, 1999; 43(3): 650 - 657.
[Abstract] [Full Text] [PDF]


Home page
Arch SurgHome page
N. K. Veeramachaneni, A. H. Harken, and C. B. Cairns
Clinical Implications of Hemoglobin as a Nitric Oxide Carrier
Arch Surg, April 1, 1999; 134(4): 434 - 437.
[Abstract] [Full Text] [PDF]


Home page
J Am Coll CardiolHome page
T. Le Tourneau, E. Van Belle, D. Corseaux, B. Vallet, G. Lebuffe, B. Dupuis, J.-M. Lablanche, E. McFadden, C. Bauters, and M. E. Bertrand
Role of nitric oxide in restenosis after experimental balloon angioplasty in the hypercholesterolemic rabbit: effects on neointimal hyperplasia and vascular remodeling
J. Am. Coll. Cardiol., March 1, 1999; 33(3): 876 - 882.
[Abstract] [Full Text] [PDF]


Home page
Arterioscler. Thromb. Vasc. Bio.Home page
S. Fang, R. V. Sharma, and R. C. Bhalla
Enhanced Recovery of Injury-Caused Downregulation of Paxillin Protein by eNOS Gene Expression in Rat Carotid Artery : Mechanism of NO Inhibition of Intimal Hyperplasia?
Arterioscler. Thromb. Vasc. Biol., January 1, 1999; 19(1): 147 - 152.
[Abstract] [Full Text] [PDF]


Home page
CirculationHome page
A. Gries, C. Bode, K. Peter, A. Herr, H. Bohrer, J. Motsch, and E. Martin
Inhaled Nitric Oxide Inhibits Human Platelet Aggregation, P-Selectin Expression, and Fibrinogen Binding In Vitro and In Vivo
Circulation, April 21, 1998; 97(15): 1481 - 1487.
[Abstract] [Full Text] [PDF]


Home page
Am. J. Physiol. Heart Circ. Physiol.Home page
R. O. Han, D. S. Ettenson, E. W. Y. Koo, and E. R. Edelman
Heparin/heparan sulfate chelation inhibits control of vascular repair by tissue-engineered endothelial cells
Am J Physiol Heart Circ Physiol, December 1, 1997; 273(6): H2586 - H2595.
[Abstract] [Full Text] [PDF]


Home page
Circ. Res.Home page
Z. Nong, M. Hoylaerts, N. Van Pelt, D. Collen, and S. Janssens
Nitric Oxide Inhalation Inhibits Platelet Aggregation and Platelet-Mediated Pulmonary Thrombosis in Rats
Circ. Res., November 19, 1997; 81(5): 865 - 869.
[Abstract] [Full Text]


Home page
Arterioscler. Thromb. Vasc. Bio.Home page
S. Heeneman, J. F.M. Smits, P. J.A. Leenders, P. M.H. Schiffers, and M. J.A.P. Daemen
Effects of Angiotensin II on Cardiac Function and Peripheral Vascular Structure During Compensated Heart Failure in the Rat
Arterioscler. Thromb. Vasc. Biol., October 1, 1997; 17(10): 1985 - 1994.
[Abstract] [Full Text]


Home page
Proc. Natl. Acad. Sci. USAHome page
C. Napoli, G. Aldini, J. L. Wallace, F. de Nigris, R. Maffei, P. Abete, D. Bonaduce, G. Condorelli, F. Rengo, V. Sica, et al.
Efficacy and age-related effects of nitric oxide-releasing aspirin on experimental restenosis
PNAS, February 5, 2002; 99(3): 1689 - 1694.
[Abstract] [Full Text] [PDF]


This Article