| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
1 Department of Molecular and Cellular Physiology, Louisiana State University Medical Center, Shreveport, Louisiana 71130-3392; and 2 Harvard Medical School, Cardiovascular Research Center, Massachusetts General Hospital, Charlestown, Massachusetts 02129-2060
 |
ABSTRACT |
|---|
 TOP
 ABSTRACT
 INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
Myocardial
ischemia and reperfusion (MI/R) initiates a cascade of
polymorphonuclear neutrophil (PMN)-mediated injury, the magnitude of
which may be influenced by the bioavailability of nitric oxide
(NO). We investigated the role of endothelial cell nitric
oxide synthase (ecNOS) in MI/R injury by subjecting wild-type and
ecNOS-deficient (
/) mice to 20 min of coronary artery
occlusion and 120 min of reperfusion. Myocardial infarct size
represented 20.9 ± 2.9% of the ischemic zone in wild-type mice,
whereas the ecNOS / mice had significantly
(P < 0.01) larger infarcts measuring 46.0 ± 3.8% of the ischemic zone. Because P-selectin is thought to
be involved with the pathogenesis of neutrophil-mediated I/R injury, we
assessed the effects of MI/R on P-selectin expression in the myocardium
of wild-type and ecNOS / mice. P-selectin expression
measured with a radiolabeled monoclonal antibody (MAb) technique after
MI/R in wild-type mice was 0.037 ± 0.009 µg MAb/g tissue, whereas ecNOS / coronary vasculature was
characterized by significantly (P < 0.05) higher P-selectin expression (0.080 ± 0.013 µg MAb/g
tissue). Histological examination of the postischemic myocardium
revealed significantly (P < 0.01)
more neutrophils in the ecNOS / (29.5 ± 2.5 PMN/field) compared with wild-type (5.0 ± 0.9 PMN/field) mice. A
similar trend in infarct size and neutrophil accumulation was observed
when wild-type and ecNOS / mice were subjected to 30 min
of ischemia and 120 min of reperfusion. These novel in vivo
findings demonstrate a cardioprotective role for ecNOS-derived NO in
the ischemic-reperfused mouse heart.
mouse; neutrophils; infarction; leukocyte adhesion molecules
| |
INTRODUCTION |
|---|
|
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
NITRIC OXIDE SYNTHASE (NOS) generates nitric oxide (NO) by converting L-arginine to L-citrulline (29). At present, there are three known isoforms of NOS designated: 1) neuronal NOS (nNOS or NOS 1), 2) inducible NOS (iNOS or NOS 2), and 3) endothelial cell NOS (ecNOS or NOS 3) (33). Previous investigations have shown the expression and function of the three different isoforms to vary with each organ and the physiological or pathophysiological state of the tissue. It is well accepted that ecNOS modulates a variety of tissue-specific events, including vasodilation (10), leukocyte-endothelial cell interactions (22), platelet adhesion (15), platelet aggregation (29, 40), microvascular permeability (21), and smooth muscle cell proliferation (12). However, the precise role of NO generated by ecNOS is not completely understood in many physiological and pathological states.
Myocardial ischemia and reperfusion (MI/R) injury is widely accepted as a stimulus for tissue destruction and possible cardiac failure (1). Accordingly, much attention has been directed toward elucidating the mechanisms of MI/R injury. Previous studies have shown this cascade of injury to be inflammatory in nature and involve interactions between circulating polymorphonuclear neutrophils (PMNs) and the coronary endothelium (30). Furthermore, PMN-mediated myocardial reperfusion injury is a sequential process involving three interdependent steps: 1) neutrophil rolling, 2) firm adhesion, and 3) transmigration (24).
Although numerous studies have investigated the possible function of NO in pathological sequelae, no area of NO physiology has attracted more attention than its involvement in I/R injury. Many studies indicate an important capacity for NO-mediated cardioprotection in MI/R injury. Previous MI/R studies have shown that NO donors attenuate I/R arrhythmias (37, 45) and myocardial infarct (Inf) size (27, 36), and improve postischemic coronary blood flow and contractile function (35, 36). Recently, it has also been demonstrated that inhaled NO protects against I/R injury (9). Similarly, other studies have shown that administration of the NO precursor L-arginine produces comparable results with respect to MI/R injury (34, 35, 46). Furthermore, studies have revealed that the benefits of L-arginine and NO donors in MI/R injury are likely related to attenuation of leukocyte-endothelial cell interactions (22, 27) and the anti-oxidant actions of NO (11, 23).
In sharp contrast to the reports demonstrating cardioprotective actions of NO in the setting of MI/R injury there are also a number of studies suggesting that NO can be cytotoxic and actually contribute to myocardial cell injury following coronary ischemia and reperfusion (31, 38, 41, 48). Administration of NOS inhibitors before ischemia has been shown to reduce myocardial infarction (38, 48) and the extent of reperfusion injury (41) following coronary ischemia and reperfusion. Finally, it has been proposed that NO formed in the coronary circulation contributes to cell signaling involved in the pathophysiology of MI/R injury (31).
Previous experimental studies on the role of NO in MI/R were
complicated by the lack of specificity and the side effects of traditional NOS inhibitors as well as the complex pharmacology of
NO-donating agents. Recently transgenic mice have been developed that
are deficient in ecNOS and therefore are incapable of endothelial cell
NO production. It has previously been reported that these mice exhibit
a complete lack of vascular reactivity to the endothelium-dependent vasodilator acetylcholine and are hypertensive (17, 43). Therefore, the
ecNOS null mouse is an excellent model system for investigation of the
potential physiological and pathophysiological actions of endothelial
cell-derived NO. In the present study, we examined the effects of
coronary artery occlusion and reperfusion in both wild-type and
ecNOS-deficient (/) mice to fully elucidate the role of
NO in MI/R injury. Specifically, we determined the extent of myocardial
cell necrosis, neutrophil accumulation, and coronary vascular
P-selectin expression in mice following acute MI/R injury.
| |
MATERIALS AND METHODS |
|---|
|
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
Transgenic mice.
ecNOS / mice were generated by Fishman and colleagues, as
described previously (17, 43). SV129 mice (Jackson Laboratories) were
utilized as the control (wild-type mice) animals for the ecNOS
/ mice. All mice utilized in the study were between 12 and 16 wk old. All experimental procedures complied with the
Guide for the Care and Use of Laboratory
Animals, DHHS Publication No. (NIH) 86-23, Revised
1985, approved by the American Physiological Society, and with federal
and state regulations.
Measurement of ecNOS mRNA by RT-PCR. Total RNA was extracted from the mouse lung using the acid guanidium-phenol-chloroform extraction method described by Chomczynski and Sacchi (4).
First-strand cDNA synthesis was performed at 42°C for 20 min using 2 µg total RNA from mouse lung in a 20-µl reaction mixture containing 50 mM Tris · HCl, pH 8.3, 75 mM KCl, 3 mM MgCl2, 10 mM dithiothreitol, 20 µM dNTP (an equal mixture of dATP, dGTP, dCTP, and dTTP), 1 µM oligo(dT), and 200 U Superscript RT (GIBCO-BRL, Gaithersburg, MD). Denaturation was performed at 65°C for 5 min, followed by primer annealing at 42°C for 5 min, before addition of the enzyme. The mouse ecNOS fragment (from 3049 to 3402) was amplified using forward primer (5'-GACTGGCATTGCACCCTTCCGG-3'), corresponds to 3049 to 3070, and reverse primer (5'-GTTGCCAGAATTCTCTGCACGG-3'), corresponds to 3402 to 3381 of the mouse ecNOS gene (14). This PCR product was cloned using PCR 2.1-TOPO Cloning Kit (gift from Invitrogen, Carlsbad, CA). The fragment of ecNOS was verified by sequencing the insert in the plasmid. The glyceraldehyde-3-phosphate dehydrogenase (GAPDH) fragment (from 300 to 614) was amplified using forward primer (5'-CACCACCATGGAGAAGGCTG-3'), corresponds to 300 to 319, and reverse primer (5'-ATGATGTTCTGGGCAGCCCC-3'), corresponds to 614 to 5595 of the rat GAPDH gene (44).Surgical procedures. Animals were anesthetized with pentobarbital sodium (100 mg/kg ip). Anesthesia was maintained via supplemental doses of pentobarbital sodium (30 mg/kg ip) as needed. A midline incision was made from the xiphoid process to the submentum. The salivary glands were separated from the midline to allow access to the trachea. A tracheotomy was then performed to facilitate breathing. A section of polyethylene 90 tubing was inserted into the animal's trachea and connected via a loose junction to a Harvard respirator (model 683 rodent respirator, Harvard Apparatus). The respirator's tidal volume was set at 1.0 ml/min, and the rate was set at 120 strokes/min and was supplemented with 100% oxygen. The right carotid artery was then cannulated with polyethylene 10 tubing to monitor hemodynamics and to facilitate the infusion of Evans blue at the end of the experiment.
After an equilibration period of 10 min, a thoracotomy was performed. Using an electrocautery (model 100, Geiger Instrument), we made an incision to the left of the sternum. The pericardial sac was then removed. Ligation of the left anterior descending (LAD) coronary artery was performed using a 7-0 silk suture attached to a BV-1 needle (Ethicon). A small piece of polyethylene tubing was used to secure the ligature without damaging the artery. The chest wall was approximated and covered with Parafilm wax paper to prevent desiccation. At the appropriate time (20 or 30 min of ischemia), the 7-0 silk ligature was cut and removed from the heart. Reperfusion was visually confirmed in all animals using a dissecting microscope. Animals that did not undergo complete LAD reperfusion were excluded from the study. At 5 min of myocardial ischemia wild-type mice received a bolus injection of epinephrine (1 µg in 50 µl saline) via the carotid artery catheter to maintain systemic blood pressure similar to that observed in ecNOS/ mice. Epinephrine (1 µg) was
administered to wild-type mice throughout the experimental protocol as
needed to maintain blood pressure at levels observed in the ecNOS
/ animals.
Myocardial P-selectin expression.
Radiolabeled P-selectin and control monoclonal antibodies (MAb) were
prepared as previously described (8). All mice were instrumented with
carotid artery and jugular vein catheters. Coronary vascular P-selectin
expression in wild-type (n = 5) and
ecNOS / (n = 3) mice was assessed following sham myocardial ischemia (20 min) and reperfusion (20 min). Additionally, expression of P-selectin
in the coronary vasculature was assessed following MI/R in wild-type
(n = 4) and ecNOS /
(n = 4) mice. All animals underwent 20 min of LAD occlusion followed by 20 min of reperfusion. At
15 min of reperfusion, radiolabeled antibodies were injected. Monoclonal radiolabeled (125I)
antibody directed against P-selectin (RB40.34, PharMingen) and a
nonbinding radiolabeled (131I)
antibody (P-23, Pharmacia-Upjohn) were slowly administered through the
jugular vein. After 5 min of circulation (20 min of reperfusion), a
50-µl plasma sample was drawn. The animal was then perfused with 15 ml of warm (pH 7.4), heparinized bicarbonate-buffered saline, while
being exsanguinated to flush the excess P-selectin MAb and nonbinding
control antibody. LAD religation was followed by infusion of ~2 ml of
1% Evans blue to delineate the ischemic zone from the nonischemic
zone. The heart was then excised and serially sectioned. The
nonischemic and ischemic zones were separated and weighed, and regional
cardiac radioactivity was measured using an automatic gamma counter
(1480 Wizard, Wallac) to determine MI/R induced P-selectin expression.
Myocardial histology.
Routine histological staining was performed on multiple sections of
midventricular cardiac sections to determine the extent of neutrophil
(PMN) infiltration. Wild-type (n = 3)
and ecNOS / (n = 3)
hearts were subjected to 20 min of myocardial ischemia and 120 min of reperfusion and stained as previously described. In additional
studies, wild-type (n = 4) and ecNOS
/ (n = 4) mouse hearts
were subjected to 30 min of LAD occlusion and 120 min of reflow and
then submitted for the aforementioned staining protocol.
Determination of area at risk and Inf size. At the conclusion of the 2-h period of reperfusion, the LAD was religated with 7-0 silk suture, and 1.2 ml of 1.0% Evans blue (Sigma) was retrogradely injected into the carotid artery catheter to delineate the in vivo area at risk (AAR).
At the end of the protocol, the heart was excised and fixed in 1.5% solution of SeaPlaque agarose gel (FMC BioProducts). After the gel solidified, the heart was sectioned perpendicular to the long axis in 1-mm portions using a McIlwain tissue chopper (Brinkmann Instruments). The 1-mm sections were placed in individual wells of a six-well cell culture plate with the basal side exposed. Each slice was then counterstained with 3.0 ml of 1.0% 2,3,5-triphenyltetrazolium chloride (Sigma) solution for 5 min at 37°C. Each slice was weighed and visualized under an Olympus SZ4045 (Olympus America) dissecting microscope equipped with a Sony charge-coupled device iris-color video camera (Sony Electronics). The left ventricular area, AAR, and area of infarction for each slice were then determined by computer planimetry using National Institutes of Health Image (v1.57) software. The size of the myocardial infarction was determined by the following previously described equation (32): weight of infarction equals (A1 × Wt1) + (A2 × Wt2) + (A3 × Wt3) + (A4 × Wt4) + (A5 × Wt5), where A is percent area of infarction by planimetry from subscripted numbers 1-5 representing sections and Wt is the weight of the same numbered sections.Statistical analyses. The data were analyzed with ANOVA and Scheffé's post hoc test. All values are reported as means ± SE. Statistical significance was set at P < 0.05.
| |
RESULTS |
|---|
|
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
Detection of ecNOS mRNA.
RT-PCR was performed on two wild-type and two ecNOS /
lungs for the detection of ecNOS mRNA. As shown in Fig.
1, the distinct bands indicate that ecNOS
mRNA is present in both wild-type animals. Conversely, the
corresponding bands are completely absent in the ecNOS /
animals. This confirms the absence of ecNOS in our ecNOS / mice.
|
Hemodynamic measurements.
Mean arterial blood pressure (MABP) and heart rate (HR) were recorded
in all groups throughout the experimental protocols. In addition, we
calculated the rate-pressure product (RPP, MABP × HR/1,000) as an
index of oxygen demand. The data for the 20 min of myocardial
ischemia and 120 min of reperfusion are reported in Table
1. Table 2
summarizes the hemodynamic data from the 30 min of myocardial
ischemia and 120 min of reperfusion group. The
ecNOS / mice exhibited significantly
(P < 0.05) higher baseline MABP and
RPP values compared with wild-type mice in both the 20- and 30-min
myocardial ischemia protocols. Also, at 60 min of reperfusion, the ecNOS / had a significantly
(P < 0.05) lower mean RPP. The remainder of the time points did not demonstrate significantly different values.
|
|
Left ventricular mass.
Chronic hypertension has been shown to result in left ventricular
hypertrophy. We measured the body weight and the left ventricular weight of each to assess the degree of left ventricular hypertrophy in
the ecNOS / mice. The heart weight (mg)-to-body weight
(g) ratio for ecNOS / mice was 3.04 ± 0.20 (n = 14) and for ecNOS /
and 2.90 ± 0.17 for wild-type animals
[P = not significant (NS)].
Myocardial P-selectin expression.
The dual-radiolabeled MAb technique was used to quantify the in vivo
coronary endothelial cell expression of P-selectin following 20 min of
coronary ischemia and 20 min of reperfusion. The data for the
sham I/R experiments revealed no difference
(P = NS) between wild-type (0.001 ± 0.001 µg MAb/g tissue) and ecNOS / (0.004 ± 0.004 µg MAb/g tissue) mouse hearts. Interestingly,
P-selectin expression (Fig. 2) in
the nonischemic zone following 20 min of myocardial ischemia
and 20 min of reperfusion was significantly (P < 0.05) higher in the ecNOS
/ hearts (0.041 ± 0.010 µg MAb/g tissue) than in
the wild-type hearts (0.010 ± 0.005 µg MAb/g tissue). Furthermore, P-selectin was expressed at significantly
(P < 0.05) higher levels in the
ischemic zone of the ecNOS / hearts compared with the
wild-type hearts. Mean P-selectin expression was 0.080 ± 0.013 µg
MAb/g tissue in the ecNOS / ischemic zone and 0.037 ± 0.009 µg MAb/g tissue in the wild-type ischemic zone.
|
Myocardial neutrophil accumulation.
Neutrophil counts within the ischemic zone determined following 20 min
of myocardial ischemia and 120 min of reperfusion are presented
in Fig.
3A. The
ecNOS / hearts contained significantly more
(P < 0.01) neutrophils than the
wild-type hearts (29.5 ± 2.5 vs. 5.0 ± 0.9 PMN/field).
|
/ hearts following 30 min of LAD
occlusion and 120 min of reflow are presented in Fig.
3B. Significantly more
(P < 0.01) neutrophils were
sequestered in the ecNOS / hearts (68.6 ± 4.0 PMN/field) compared with the wild-type control hearts (24.7 ± 3.4 PMN/field).
Myocardial AAR and Inf size.
Summary data for AAR and Inf size following 20 min of coronary
occlusion and 120 min of reflow are shown in Fig.
4A.
Both groups of animals experienced similar-sized ischemic zones per left ventricle (LV) (ecNOS / AAR equals 46.0 ± 3.8%
of LV; wild type AAR equals 43.3 ± 2.9% of LV;
P = NS). Inf size was 20.9 ± 3.4%
of the AAR in wild-type mice and 46.7 ± 4.0% of the AAR in ecNOS
/ mice (P < 0.01).
|
/ AAR equals 54.4 ± 3.2%; wild-type AAR equals 46.6 ± 2.1%), the portion of this area
rendered necrotic (Inf/AAR) was significantly more extensive
(P < 0.01) in the ecNOS
/ hearts (58.4 ± 2.9%) compared with the wild-type
controls (33.0 ± 7.5%).
| |
DISCUSSION |
|---|
|
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
There are a number of salient findings revealed in this study. The most
impressive finding of this study is the extreme disparity in myocardial
infarct size between a wild-type mouse and a mouse that does not
produce NO from ecNOS. This dichotomy between the wild-type and ecNOS
/ mice was observed after both 20 and 30 min of
myocardial ischemia followed by 120 min of reperfusion. This
clearly emphasizes the cardioprotection afforded by constitutive generation of NO. The infarct data alone indicate that NO seems to be a
cornerstone of the intrinsic defenses of the body against MI/R-induced
myocardial cell injury. The results of the present study agree with and
extend previous observations of exogenous NO-exerting cardioprotective
effects (19, 27, 37), whereas NOS inhibitors exacerbated myocardial
injury following ischemia and reperfusion (16, 35).
Using gene-targeted knockout mice, we focused on the role of
physiological levels of NO production rather than administration of NO
donors or NOS inhibitors. Our approach avoided the possible hazards
associated with these previous studies because the use of either of
these methods involves determining the appropriate agent, dose, route
of administration, half-life, and time of administration. In addition,
NOS inhibitors can range from isoform selective to entirely
nonselective. In the present study, we have eliminated these precarious
steps with the use of genetically altered mice. However, there are some
considerations to be made concerning the use of these mice. Although
the measured parameters of oxygen demand did not differ significantly
between ecNOS / and wild-type mice during
ischemia, there may be constitutive changes in the cardiac
phenotype of the ecNOS / mice due to long-standing
hypertension. In addition, the vasoreactivity of the ecNOS
/ mice may be impaired. Consequently, we cannot
completely rule out the possibility of delayed or no reflow.
The present study also demonstrates increased myocardial necrosis
accompanied by enhanced neutrophil accumulation in the
ischemic-reperfused myocardium of the ecNOS / mice. This
investigation confirmed a large disparity between wild-type and ecNOS
/ mice in terms of neutrophil infiltration after both 20 and 30 min of coronary occlusion and 120 min of
reperfusion. These data implicate neutrophils as a key
mediator for the exaggerated myocardial injury resulting from
ischemia and reperfusion in the ecNOS / mice.
There is a large body of evidence suggesting that MI/R injury is
largely an inappropriate immune response that is mediated by
neutrophils (7, 30). It has previously been demonstrated that
neutrophils contribute to MI/R injury in a variety of animals models
and in humans (18, 20). In addition, anti-neutrophils agents are cardioprotective in the setting of myocardial reperfusion injury (30).
Neutrophil-mediated myocardial injury is dependent on the interaction
of adhesion glycoproteins expressed on the surface of circulating
neutrophils (L-selectin, P-selectin glycoprotein ligand-1,
and CD11/CD18) with the counter receptors expressed on the surface of
the coronary endothelium [P-selectin, E-selectin, and
intercellular adhesion molecule 1 (ICAM-1)] (2, 7,
24). NO released by the endothelium has been shown to
inhibit the surface expression of a number of endothelial cell adhesion
molecules, including P-selectin (5, 13), E-selectin
(6), vascular cell adhesion molecule 1 (42), and ICAM-1 (6, 42). In
addition, NO has also been shown to inhibit the activation of nuclear
transcription factor 
B, which is thought to regulate numerous
inflammatory and immune responses involving endothelial cell adhesion
molecule expression (39). Moreover, the antiadhesive actions of NO are thought to be in part related to the anti-oxidant actions of NO in the
microcirculation (11).
The upregulation of P-selectin following MI/R is understood to be an early and necessary step for neutrophil tethering to the coronary endothelium (24, 28). Consequently, the degree of P-selectin expression determines the abundance of rolling neutrophils that may eventually adhere to the endothelium, extravasate into the tissue, and thereby mediate necrosis. Previous investigations have shown that inhibition of P-selectin with MAb (25, 47) directed against P-selectin or a soluble carbohydrate ligand (3, 26) attenuated myocardial necrosis following MI/R. The P-selectin expression data from the present study extend these observations further. We observed that the absence of ecNOS (as confirmed by RT-PCR) resulted in a significant increase in coronary P-selectin expression following MI/R. This indicates that NO generated by ecNOS at least partially governs P-selectin expression subsequent to ischemia and reperfusion of the myocardium.
In conclusion, the present study clearly reinforces previous studies that suggest the vital role that endothelial cell-derived NO plays in vascular homeostasis within the coronary circulation. Abolition of ecNOS dramatically exacerbated the extent of myocardial reperfusion injury following acute coronary artery ischemia and reperfusion. In addition, the level of coronary P-selectin expression and PMN infiltration into the ischemic-reperfused myocardium was markedly increased in mice lacking the gene for ecNOS. The results of this study provide further support for the anti-neutrophil actions of NO in the setting of acute inflammation and also suggest that NO may be of tremendous value for the treatment of MI/R injury.
| |
ACKNOWLEDGEMENTS |
|---|
We thank DeRoyal Surgical (Powell, TN) and Ethicon Surgical (Somerville, NJ) for the generous donation of surgical supplies.
| |
FOOTNOTES |
|---|
We acknowledge the expert assistance of Dr. Alexander Minchenko for performing the RT-PCR experiments.
This research was supported by the National Institute of Diabetes and Digestive and Kidney Diseases Grant PO1-DK-43785 to D. N. Granger and by Grant JDF-195065 from the Juvenile Diabetes Foundation to D. J. Lefer.
The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.
Address for reprint requests and other correspondence: D. J. Lefer, Dept. of Molecular and Cellular Physiology, LSU Medical Center, 1501 Kings Highway, Shreveport, Louisiana 71130 (E-mail: dlefer{at}lsumc.edu).
Received 27 August 1998; accepted in final form 4 January 1999.
| |
REFERENCES |
|---|
|
TOP
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
1.
Braunwald, E.,
and
R. A. Kloner.
Myocardial reperfusion: a double edged sword.
J. Clin. Invest.
76:
1713-1719,
1985.
2.
Bruehl, R. E.,
K. L. Moore,
D. E. Lorant,
N. Borregaard,
G. A. Zimmerman,
R. P. McEver,
and
D. F. Bainton.
Leukocyte activation induces surface redistribution of P-selectin glycoprotein ligand-1.
J. Leukoc. Biol.
61:
489-499,
1997[Abstract].
3.
Buerke, M.,
A. S. Weyrich,
Z. Zheng,
F. C. Gaeta,
M. J. Forrest,
and
A. M. Lefer.
Sialyl Lewisx-containing oligosaccharide attenuates myocardial reperfusion injury in cats.
J. Clin. Invest.
93:
1140-1148,
1994.
4.
Chomczynski, P.,
and
N. Sacchi.
Single-step method of RNA isolation by acid guanidium thiocyanate-phenol-chloroform extraction.
Anal. Biochem.
162:
156-159,
1987[Medline].
5.
Davenpeck, K. L.,
T. W. Gauthier,
and
A. M. Lefer.
Inhibition of endothelial-derived nitric oxide promotes P-selectin expression and actions in the rat microcirculation.
Gastroenterology
107:
1050-1058,
1994[Medline].
6.
DeCaterina, R.,
P. Libby,
H.-B. Peng,
V. J. Thannickol,
T. B. Rajavashisth,
M. A. Gimbrone,
W. S. Shin,
and
J. K. Liao.
Nitric oxide decreases cytokine-induced endothelial activation. NO selectively reduces endothelial expression of adhesion molecules and proinflammatory cytokines.
J. Clin. Invest.
96:
60-68,
1995.
7.
Entman, M. L.,
L. H. Michael,
R. D. Rossen,
W. J. Dreyer,
D. C. Anderson,
and
C. W. Smith.
Inflammation in the course of early myocardial ischemia.
FASEB J.
5:
2529-2537,
1991[Abstract].
8.
Eppihimer, M. J.,
J. Russell,
D. C. Anderson,
B. A. Wolitzky,
and
D. N. Granger.
Endothelial cell adhesion molecule expression in gene-targeted mice.
Am. J. Physiol.
273 (Heart Circ. Physiol. 42):
H1903-H1908,
1997
9.
Fox-Robichaud, A.,
D. Payne,
S. U. Hasan,
L. Ostrovsky,
T. Fairhead,
P. Reinhardt,
and
P. Kubes.
Inhaled NO as a viable antiadhesive therapy for ischemia/reperfusion injury of distal microvascular beds.
J. Clin. Invest.
101:
2497-2505,
1998[Medline].
10.
Furchgott, R. F.,
and
J. V. Zawadzki.
The obligatory role of endothelial cells on the relaxation of arterial smooth muscle by acetylcholine.
Nature
288:
373-376,
1980[Medline].
11.
Gaboury, J.,
R. C. Woodman,
D. N. Granger,
P. Reinhardt,
and
P. Kubes.
Nitric oxide prevents leukocyte adherence: role of superoxide.
Am. J. Physiol.
265 (Heart Circ. Physiol. 34):
H862-H867,
1993
12.
Garg, U. C.,
and
A. Hassid.
Nitric oxide generating vasodilators and 8-bromo-cyclic GMP inhibit mitogenesis and proliferation of cultured rat vascular smooth muscle cells.
J. Clin. Invest.
83:
1974-1977,
1989.
13.
Gauthier, T. W.,
K. L. Davenpeck,
and
A. M. Lefer.
Nitric oxide attenuates leukocyte-endothelial interaction via P-selectin in splanchnic ischemia-reperfusion.
Am. J. Physiol.
267 (Gastrointest. Liver Physiol. 30):
G562-G568,
1994
14.
Gnanapandithen, K.,
Z. Chen,
C. L. Kau,
R. M. Gorczynski,
and
P. A. Marsden.
Cloning and characterization of murine endothelial constitutive nitric oxide synthase.
Biochim. Biophys. Acta
1308:
103-106,
1996[Medline].
15.
Groves, P. H.,
M. J. Lewis,
H. A. Cheadle,
and
W. J. Penny.
SIN-1 reduces platelet adhesion and platelet thrombus formation in a porcine model of balloon angioplasty.
Circulation
87:
590-597,
1993
16.
Hasebe, N.,
Y.-T. Shen,
and
S. F. Vatner.
Inhibition of endothelium-derived relaxing factor enhances myocardial stunning in conscious dogs.
Circulation
88:
2862-2871,
1993
17.
Huang, P. L.,
Z. Huang,
H. Mashimo,
K. D. Bloch,
M. A. Moskowitz,
J. A. Bevan,
and
M. C. Fishman.
Hypertension in mice lacking the gene for endothelial nitric oxide synthase.
Nature
377:
239-242,
1995
18.
Ikeda, H.,
Y. Takajo,
K. Ichiki,
T. Ueno,
S. Maki,
T. Noda,
K. Sugi,
and
T. Imaizumi.
Increased soluble form of P-selectin in patients with unstable angina.
Circulation
92:
1693-1696,
1995
19.
Johnson, G., III,
P. S. Tsao,
and
A. M. Lefer.
Cardioprotective effects of authentic nitric oxide in myocardial ischemia with reperfusion.
Crit. Care Med.
19:
244-252,
1991[Medline].
20.
Kakita, K.,
H. Ogawa,
H. Yasue,
T. Sakamoto,
H. Suefuji,
H. Sumida,
and
K. Okumura.
Soluble P-selectin is released into the coronary circulation after coronary spasm.
Circulation
92:
1726-1730,
1995
21.
Kubes, P., and D. N. Granger. Nitric oxide
modulates microvascular permeability.
Circulation 611-615, 1992.
22.
Kubes, P.,
M. Suzuki,
and
D. N. Granger.
Nitric oxide: an endogenous modulator of leukocyte adhesion.
Proc. Natl. Acad. Sci. USA
88:
4651-4655,
1991
23.
Kurose, I.,
R. Wolf,
M. B. Grisham,
T. Y. Aw,
R. D. Specian,
and
D. N. Granger.
Microvascular responses to inhibition of nitric oxide production. Role of active oxidants.
Circ. Res.
76:
30-39,
1995
24.
Lasky, L. A.
Selectins: interpreters of cell-specific carbohydrate information during inflammation.
Science
258:
964-969,
1992
25.
Lefer, D. J.,
D. M. Flynn,
and
A. J. Buda.
Effects of a monoclonal antibody directed against P-selectin after myocardial ischemia and reperfusion.
Am. J. Physiol.
270 (Heart Circ. Physiol. 39):
H88-H98,
1996
26.
Lefer, D. J.,
D. M. Flynn,
M. L. Phillips,
M. Ratcliffe,
and
A. J. Buda.
A novel sialyl Lewisx analog attenuates neutrophil accumulation and myocardial necrosis after ischemia and reperfusion.
Circulation
90:
2390-2401,
1994
27.
Lefer, D. J.,
K. Nakanishi,
W. E. Johnston,
and
J. Vinten-Johansen.
Antineutrophil and myocardial protecting actions of a novel nitric oxide donor after acute myocardial ischemia and reperfusion of dogs.
Circulation
88:
2337-2350,
1993
28.
Lorant, D. E.,
M. K. Topham,
R. E. Whatley,
R. P. McEver,
T. M. McIntyre,
S. M. Prescott,
and
G. A. Zimmerman.
Inflammatory roles of P-selectin.
J. Clin. Invest.
92:
559-570,
1993.
29.
Loscalzo, J.,
and
G. Welch.
Nitric oxide and its role in the cardiovascular system.
Prog. Cardiovasc. Dis.
38:
87-104,
1995[Medline].
30.
Lucchesi, B. R.
Modulation of leukocyte-mediated myocardial reperfusion injury.
Annu. Rev. Physiol.
84:
400-411,
1991.
31.
Maulik, N.,
D. T. Engelman,
M. Watanabe,
R. M. Engelman,
G. Maulik,
G. A. Cordis,
and
D. K. Das.
Nitric oxide signaling in the ischemic heart.
Cardiovasc. Res.
30:
593-601,
1995[Medline].
32.
Michael, L. H.,
M. L. Entman,
C. J. Hartley,
K. A. Youker,
J. Zhu,
S. R. Hall,
H. K. Hawkins,
K. Berens,
and
C. M. Ballantyne.
Myocardial ischemia and reperfusion: a murine model.
Am. J. Physiol.
269 (Heart Circ. Physiol. 38):
H2147-H2154,
1995
33.
Michel, T.,
and
O. Feron.
Nitric oxide synthases: which, where, how, and why?
J. Clin. Invest.
100:
2146-2152,
1997[Medline].
34.
Nakanishi, K. J.,
J. Vinten-Johansen,
D. J. Lefer,
Z. Zhao,
W. C. Fowler III,
D. S. McGee,
and
W. E. Johnston.
Intracoronary L-arginine during reperfusion improves endothelial function and reduces infarct size.
Am. J. Physiol.
263 (Heart Circ. Physiol. 32):
H1650-H1658,
1992
35.
Pabla, R.,
A. J. Buda,
D. M. Flynn,
S. A. Blesse,
A. M. Shin,
M. J. Curtis,
and
D. J. Lefer.
Nitric oxide attenuates neutrophil-mediated myocardial contractile dysfunction after ischemia and reperfusion.
Circ. Res.
78:
65-72,
1996
36.
Pabla, R.,
A. J. Buda,
D. M. Flynn,
D. B. Salzberg,
and
D. J. Lefer.
Intracoronary nitric oxide improves postischemic coronary blood flow and myocardial contractile function.
Am. J. Physiol.
269 (Heart Circ. Physiol. 38):
H1113-H1121,
1995
37.
Pabla, R.,
and
M. J. Curtis.
Effects of NO modulation on cardiac arrhythmias in the rat isolated heart.
Circ. Res.
77:
984-992,
1995
38.
Patel, V. C.,
D. M. Yellon,
K. J. Singh,
G. H. Neild,
and
R. G. Woolfson.
Inhibition of nitric oxide limits infarct size in the in situ rabbit heart.
Biochem. Biophys. Res. Commun.
194:
234-238,
1993[Medline].
39.
Peng, H. B.,
P. Libby,
and
J. K. Liao.
Induction and stabilization of I kappa B alpha by nitric oxide mediates inhibition of NF-kappa B.
J. Biol. Chem.
270:
14214-14219,
1995
40.
Radomski, M. W.,
R. M. J. Palmer,
and
S. Moncada.
The anti-aggregating properties of vascular endothelium: interactions between prostacyclin and nitric oxide.
Br. J. Pharmacol.
92:
639-644,
1987[Medline].
41.
Schultz, R.,
and
R. Wambolt.
Inhibition of nitric oxide synthesis protects the isolated working rabbit heart from ischaemia-reperfusion injury.
Cardiovasc. Res.
30:
432-439,
1995[Medline].
42.
Shin, W. S.,
Y. H. Hong,
H. B. Peng,
R. de Caterina,
P. Libby,
and
J. K. Liao.
Nitric oxide attenuates vascular smooth muscle cell activation by interferon-gamma. The role of constitutive NF-kappa B activity.
J. Biol. Chem.
271:
11317-11324,
1996
43.
Steudel, W.,
F. Ichinose,
P. L. Huang,
W. E. Hurford,
R. C. Jones,
J. A. Bevan,
M. C. Fishman,
and
W. M. Zapol.
Pulmonary vasoconstriction and hypertension in mice with targeted disruption of the endothelial nitric oxide synthase (NOS) gene.
Circ. Res.
81:
34-41,
1997
44.
Tso, J. Y.,
X. H. Sun,
T. Kao,
K. S. Reece,
and
R. Wu.
Isolation and characterization of rat human glyceraldehyde-3-phosphate dehydrogenase cDNAs: genomic complexity and molecular evolution of the gene.
Nucleic Acids Res.
13:
2485-2502,
1985
45.
Wainwright, C. L., and P. A. Martorana.
Pirsidomine, a novel nitric oxide donor, suppresses ischemic
arrhythmias in anesthetized pigs. J. Cardiovasc.
Pharmacol. 22, Suppl. 7: S44-S50,
1993.
46.
Weyrich, A. S.,
X.-L. Ma,
and
A. M. Lefer.
The role of L-arginine in ameliorating reperfusion injury after myocardial ischemia in the cat.
Circulation
86:
279-288,
1993
47.
Weyrich, A. S.,
X. Y. Ma,
D. J. Lefer,
K. H. Albertine,
and
A. M. Lefer.
In vivo neutralization of P-selectin protects feline heart and endothelium in myocardial ischemia and reperfusion injury.
J. Clin. Invest.
91:
2620-2629,
1993.
48.
Woolfson, R. G.,
V. C. Patel,
G. H. Neild,
and
D. M. Yellon.
Inhibition of nitric oxide synthesis reduces infarct size by an adenosine-dependent mechanism.
Circulation
91:
1545-1551,
1995
This article has been cited by other articles:
|
|
 |
|
  U. Landmesser, K. C. Wollert, and H. Drexler Potential novel pharmacological therapies for myocardial remodelling Cardiovasc Res, December 15, 2008; (2008) cvn317v2. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Kobayashi, Y. Tanoue, M. Eto, H. Baba, S. Kimura, S. Oda, K. Taniguchi, and R. Tominaga A Rho-kinase inhibitor improves cardiac function after 24-hour heart preservation J. Thorac. Cardiovasc. Surg., December 1, 2008; 136(6): 1586 - 1592. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Matsuhisa, H. Otani, T. Okazaki, K. Yamashita, Y. Akita, D. Sato, A. Moriguchi, H. Imamura, and T. Iwasaka Angiotensin II type 1 receptor blocker preserves tolerance to ischemia-reperfusion injury in Dahl salt-sensitive rat heart Am J Physiol Heart Circ Physiol, June 1, 2008; 294(6): H2473 - H2479. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. J. Leary, S. Rajasekaran, R. R. Morrison, E. I. Tuomanen, T. K. Chin, and P. A. Hofmann A cardioprotective role for platelet-activating factor through NOS-dependent S-nitrosylation Am J Physiol Heart Circ Physiol, June 1, 2008; 294(6): H2775 - H2784. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Q. Wang, R. V. Donthi, J. Wang, A. J. Lange, L. J. Watson, S. P. Jones, and P. N. Epstein Cardiac phosphatase-deficient 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase increases glycolysis, hypertrophy, and myocyte resistance to hypoxia Am J Physiol Heart Circ Physiol, June 1, 2008; 294(6): H2889 - H2897. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Fu, Z. Wang, W. L. Chen, P. K. Moore, and Y. Z. Zhu Cardioprotective effects of nitric oxide-aspirin in myocardial ischemia-reperfused rats Am J Physiol Heart Circ Physiol, September 1, 2007; 293(3): H1545 - H1552. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. A. Kaufmann, C. Lewis, A. Xie, A. Mirza-Mohd, and J. R. Lindner Detection of recent myocardial ischaemia by molecular imaging of P-selectin with targeted contrast echocardiography Eur. Heart J., August 2, 2007; 28(16): 2011 - 2017. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. K. Means, C.-Y. Xiao, Z. Li, T. Zhang, J. H. Omens, I. Ishii, J. Chun, and J. H. Brown Sphingosine 1-phosphate S1P2 and S1P3 receptor-mediated Akt activation protects against in vivo myocardial ischemia-reperfusion injury Am J Physiol Heart Circ Physiol, June 1, 2007; 292(6): H2944 - H2951. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Hazarika, M. R. Van Scott, and R. M. Lust Severity of myocardial injury following ischemia-reperfusion is increased in a mouse model of allergic asthma Am J Physiol Heart Circ Physiol, January 1, 2007; 292(1): H572 - H579. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. W. Elrod, J. J. M. Greer, and D. J. Lefer Sildenafil-mediated acute cardioprotection is independent of the NO/cGMP pathway Am J Physiol Heart Circ Physiol, January 1, 2007; 292(1): H342 - H347. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. R. Duranski, J. W. Elrod, J. W. Calvert, N. S. Bryan, M. Feelisch, and D. J. Lefer Genetic overexpression of eNOS attenuates hepatic ischemia-reperfusion injury Am J Physiol Heart Circ Physiol, December 1, 2006; 291(6): H2980 - H2986. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. O. Weinberg, M. Scherrer-Crosbie, M. H. Picard, B. A. Nasseri, C. MacGillivray, J. Gannon, Q. Lian, K. D. Bloch, and R. T. Lee Rosuvastatin reduces experimental left ventricular infarct size after ischemia-reperfusion injury but not total coronary occlusion Am J Physiol Heart Circ Physiol, April 1, 2005; 288(4): H1802 - H1809. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Altay, E. R. Gonzales, T. S. Park, and J. M. Gidday Cerebrovascular inflammation after brief episodic hypoxia: modulation by neuronal and endothelial nitric oxide synthase J Appl Physiol, March 1, 2004; 96(3): 1223 - 1230. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Vinten-Johansen Involvement of neutrophils in the pathogenesis of lethal myocardial reperfusion injury Cardiovasc Res, February 15, 2004; 61(3): 481 - 497. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. P. Jones, J. J. M. Greer, A. K. Kakkar, P. D. Ware, R. H. Turnage, M. Hicks, R. van Haperen, R. de Crom, S. Kawashima, M. Yokoyama, et al. Endothelial nitric oxide synthase overexpression attenuates myocardial reperfusion injury Am J Physiol Heart Circ Physiol, January 1, 2004; 286(1): H276 - H282. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. A. Wink, K. M. Miranda, T. Katori, D. Mancardi, D. D. Thomas, L. Ridnour, M. G. Espey, M. Feelisch, C. A. Colton, J. M. Fukuto, et al. Orthogonal properties of the redox siblings nitroxyl and nitric oxide in the cardiovascular system: a novel redox paradigm Am J Physiol Heart Circ Physiol, December 1, 2003; 285(6): H2264 - H2276. [Abstract] [Full Text] [PDF]
|
|
|
