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Circulation. 1997;96:3314-3320

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(Circulation. 1997;96:3314-3320.)
© 1997 American Heart Association, Inc.

Articles

Increased Formation of the Isoprostanes IPF2{alpha}-I and 8-Epi-Prostaglandin F2{alpha} in Acute Coronary Angioplasty

Evidence for Oxidant Stress During Coronary Reperfusion in Humans

Muredach P. Reilly, MB; Norman Delanty, MB; Louis Roy, MD; Joshua Rokach, PhD; Peter O. Callaghan, MB; Peter Crean, MD; John A. Lawson; ; Garret A. FitzGerald, MD

From The Center for Experimental Therapeutics, University of Pennsylvania (M.P.R., N.D., J.A.L., G.A.F.); Institute of Cardiology, Hopital Laval, Quebec City, Canada (L.R.); Claude Pepper Institute and Department of Chemistry, Florida Institute of Technology (J.R.), Melbourne; and Department of Cardiology, St James Hospital, Dublin, Ireland (P.O.C., P.C.).


*    Abstract
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 *Abstract
down arrowIntroduction
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 down arrowResults
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Background The role of oxidant stress in cardiac ischemia/reperfusion injury in humans remains controversial. This is due, in part, to the limitations of available indices of oxidant stress in vivo. Isoprostanes are stable, free radical–catalyzed products of arachidonic acid. We assessed their formation in patients undergoing coronary reperfusion via percutaneous transluminal coronary angioplasty (PTCA).

Methods and Results We developed specific, mass spectrometry assays for two structurally distinct F2 isoprostanes, 8-epi-PGF2{alpha} and IPF2{alpha}-I. Urine samples for isoprostane determination were collected in patients undergoing coronary arteriography (n=11), elective PTCA (n=15), and angiography after thrombolysis for acute myocardial infarction (MI) (n=10). Urinary levels (pmol/mmol creatinine) of both isoprostanes were markedly increased from baseline in the first 6 hours after PTCA for acute MI (105±17.8 versus 230±66 for 8-epi-PGF2{alpha} [P=.009] and 466±91 versus 833±153 for IPF2{alpha}-I [P=.001]) and returned toward preprocedural values by 24 hours (122±18 for 8-epi-PGF2{alpha} and 457±102 for IPF2{alpha}-I). There was a slight increase in urinary 8-epi-PGF2{alpha} levels (64.7±9.5 versus 84.9±10.6; P=.02) after diagnostic coronary arteriography and elective PTCA (88.7±7.5 versus 114.3±16.1; P=.01). A striking correlation was observed (r=.68, P<.0001; n=33) between urinary 8-epi-PGF2{alpha} and IPF2{alpha}-I levels in patients receiving thrombolytic agents for acute MI.

Conclusions Urinary F2 isoprostane levels are elevated in patients after treatments resulting in reperfusion for acute MI. These findings provide evidence consistent with increased oxidant stress in vivo in this setting. Measurement of urinary isoprostanes may offer a noninvasive approach to the assessment of oxidant stress and the efficacy of antioxidant therapies in these syndromes.


Key Words: oxidant stress • lipids • angioplasty


*    Introduction
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 up arrowAbstract
 *Introduction
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 down arrowDiscussion
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The role of oxidant stress in the pathophysiology of human disease remains controversial.1 This is particularly true in the case of cardiac ischemia reperfusion.2,3 Despite evidence of a relationship between free radical generation and myocardial dysfunction in animal models,4,5 little evidence of such a relationship exists in humans. Thus, in patients with acute MI who are treated with thrombolytic drugs or PTCA, it remains unclear whether interventions aimed at suppressing free radical generation can improve clinical outcome. Attempts to address this possibility are constrained, in part, by the limitations intrinsic to available indices of oxidant stress in vivo.6,7 Traditional approaches have targeted nonspecific or unstable products of such reactions8,9 or have relied on spin adduct formation or lipoprotein oxidizability induced ex vivo.10,11 More recently, a number of novel assays of more stable markers of oxidant stress have been described,12,13 including the estimation of a family of stable end products of lipid peroxidation, the isoeicosanoids.14,15

Arachidonic acid is subject to oxidative modification, resulting in isomeric species analogous to the traditional enzymatic products of this fatty acid.16 Thus, IsoPs include isomers of the D, E, and F PGs.17 F2 IsoPs, isomers of PGF2{alpha}, are divided into four classes according to the site of free radical attack on arachidonic acid (Fig 1Down).18 They have been detected in human urine and plasma,14,15 and a number have been ascribed biological activities in vitro.19,20 We developed specific and sensitive assays using GC/MS for two distinct F2 IsoPs, 8-epi-PGF2{alpha} (class IV)21 and IPF2{alpha}-I (class I)22 and are exploring their potential use as specific, chemically stable, noninvasive indices of free radical generation in vivo. We previously reported finding increased formation of 8-epi-PGF2{alpha} in several syndromes of oxidant stress in humans, such as cigarette smoking, acetaminophen poisoning, and coronary reperfusion with thrombolytic drugs.15,23,24



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  		Figure 1. Arachidonic acid is subject to attack in situ on membrane phospholipids by free radicals (FR), leading to IsoP generation. Alternatively, it may be metabolized enzymatically, after phospholipase-dependent cleavage from cell membranes, by COXs to produce PGs. The F2 IsoP, isomers of PGF2{alpha}, can be divided into four classes: 8-epi-PGF2{alpha} belongs to class IV, and IPF2{alpha}-I belongs to class I. 8-Epi PGF2{alpha} is a minor byproduct of the COX pathways in vitro. However, in parallel experiments, IPF2{alpha}-I is not generated in this manner.

ROS are thought to mediate the reperfusion injury that characterizes myocardial stunning in animal models of coronary occlusion/reperfusion. Indeed, suppression of ROS generation in such models has resulted in a marked attenuation of myocardial dysfunction.4,5 Syndromes of reperfusion injury in humans include the regional myocardial stunning seen in some patients after acute revascularization for MI and the global myocardial reperfusion injury that may follow CABG.25,26 Furthermore, increased free radical generation has been reported in both of these settings.27–32 However, the effect of administration of antioxidant drugs during reperfusion for MI has been disappointing.33,34 It is unclear whether this reflected a true lack of efficacy, a deficiency of trial design, or inadequate dose finding. Remarkably, the most appropriate antioxidant dosing regimen for vitamins C and E in patients with cardiovascular disease remains to be determined.

We recently demonstrated increased generation of 8-epi-PGF2{alpha} in a number of syndromes of cardiac ischemia/reperfusion.24 The results of present study extend these observations by demonstrating a relationship between urinary IsoP generation and reperfusion in patients undergoing acute revascularization for MI. Furthermore, similar results are obtained with measurement of two distinct F2 IsoPs. These results reinforce the likelihood that urinary IsoPs may represent a time-integrated, noninvasive index of free radical generation in syndromes of cardiac reperfusion. Measurement of IsoPs may be used to guide the development and evaluation of rational antioxidant regimens in this setting.


*    Methods
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 *Methods
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Clinical Studies
Patients were enrolled at Saint James' Hospital, Dublin, Ireland, and at Hopital Laval, Quebec, Canada. All studies were approved by the institutional review boards of the respective hospitals, and informed consent was obtained from the volunteers participating in the investigation. Three studies were performed. The first two were conducted at Saint James' Hospital. The patient mix reflected that of the hospital referral base; all were Caucasian, and socioeconomic classes 3 through 5 predominated (15 of 26). In the first, urinary excretion of F2 IsoPs was characterized in 11 clinically stable patients (age, 46 to 70 years; 7 men and 4 women) undergoing diagnostic coronary arteriography. Clinical and angiographic details of these patients are provided in the TableDown. Four patients had previously documented coronary disease, 10 had symptoms of angina, and all had positive stress tests. Eight patients were taking aspirin, and 2 were cigarette smokers. Seven patients had significant coronary disease (>50% lesion in at least one vessel) at coronary arteriography. Three serial 6-hour urine samples were collected from these patients, one before and two after (0 to 6 hours and 6 to 12 hours) angiography. There were no complications related to the procedure.


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  		Table 1. Clinical and Angiographic Characteristics of Patients Undergoing Diagnostic Coronary Angiography, Elective PTCA, and Angiography After Acute MI

In the second study, urinary IsoP excretion was assessed in 15 patients (age, 45 to 72 years; 11 men and 4 women) undergoing elective PTCA. Clinical and angiographic details of these patients are summarized in the TableUp. All patients had undergone diagnostic coronary catheterization at the same hospital within the previous month and had symptoms typical of angina. All patients had one-vessel coronary artery disease. All patients received chronic aspirin therapy before the procedure, and 2 were current smokers. Concurrent heparin was administered to all patients and continued until the morning after the procedure, when femoral lines were removed. There were no acute vessel closures in this study group, and no patient experienced complicating MI or significant bleeding. There were no stent implantations. Serial 6-hour urine samples were collected from these patients in a similar protocol to that in the first study.

The third study was performed at Hopital Laval. IsoP generation was examined in a cohort of patients who were sequentially randomized to the PRIME study35 at this center and in whom serial urinary samples could be collected. This study was a multicenter, randomized, controlled, dose-finding trial of Efegatran (a specific thrombin inhibitor) in patients receiving thrombolysis for acute MI. All patients had typical chest pain within 12 hours of arrival at the emergency department (ED) with ST-segment elevation on their initial ECG and no contraindication to thrombolytics. Patients received oral aspirin (325 mg) and front-loaded IV rt-Pa (15-mg bolus, 0.75 mg/kg over the following 30 minutes, and 0.5 mg/kg over a final 60 minutes) and underwent coronary angiography {approx}90 minutes after administration of lytic drug. At this time, PTCA was performed if the culprit vessel was closed or had TIMI grade flow of <2. Ten patients (age, 37 to 70 years; 10 men) of 38 who had been randomized during a 12-month period, had adequate urine sampling performed. This involved the collection of a "spot" urine sample in the ED before undergoing cardiac catheterization and three sequential 6-hour urine collections after the performance of angiography or PTCA. Seven of the 10 patients underwent PTCA in this setting; 5 patients had an occluded vessel, and 2 had TIMI grade 1 flow. Clinical and angiographic details of these patients are given in the TableUp.

IsoP Analysis
Urine samples were collected into polyethylene bottles containing 0.1% of the antioxidant, butylated hydroxyanisole. They were kept refrigerated during the collection period, after which they were immediately aliquoted and stored at -70°C. Samples were bulk shipped on dry ice to the Center for Experimental Therapeutics, University of Pennsylvania, where IsoP analysis was performed.

8-Epi PGF2{alpha} was quantified by GC/MS as previously described.21 Urinary IPF2{alpha}-I was analyzed using similar techniques with the following modifications.22 The internal standard was tetradeuterated(17,17,18,18) IPF2{alpha}-I. After the addition of the internal standard to a 100-µL urine sample, 250 mg of 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide methiodide was added and allowed to stand for 30 minutes at room temperature. The sample was then extracted on a Rapid Trace solid-phase extraction workstation using a C18 SPE cartridge. The solvent program included washes with 1 mL each of H2O and 25% ethanol/H2O followed by elusion with 1 mL of ethyl acetate. After drying under N2, the sample was dissolved in 10 µL of methanol and 15% KOH/H2O and allowed to stand for 30 minutes at room temperature. After the addition of 200 µL of 0.5 N HCl, the sample was extracted with 1 mL of ethyl acetate and dried under N2. Formation of the pentafluorobenzyl ester and subsequent thin-layer chromatographic purification were carried out as for 8-epi-PGF2a. For GC/MS analysis, the trimethylsilyl derivative, as opposed to the tert-butyldimethylsilyl ester, was used. The GC was programmed to go from 190°C to 320°C at 20°C/min; retention time was {approx}8.5 minutes. Ions monitored were m/z 569 and 573. Quantification of both IsoPs was performed using peak area ratio. Urinary creatinines were determined by the alkaline picrate (Jaffe) reaction,36 and urinary IsoP levels were expressed as picomoles per millimole (pmol/mmol) of creatinine.

Blood samples were collected for serum creatine phosphokinase levels in patients with acute MI at 6, 12, and 24 hours after admission. Levels were assayed in the clinical chemistry laboratory of the Hopital Laval.

Statistical Analysis
A paired t test was used when two samples from the same patients were being compared. More than two samples were compared with the use of a one-way ANOVA and, if significant differences occurred, by Duncan's multiple range tests to assess where the differences lay. Only preplanned comparisons were tested, and significance levels were corrected for number of comparisons made. Relationships between two continuous variables were assessed by regression analysis using the Pearson correlation coefficient. All data are expressed as mean±SEM, and differences were considered significant at a value of P<.05.


*    Results
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 *Results
down arrowDiscussion
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Urinary 8-epi-PGF2{alpha} excretion increased slightly after the performance of diagnostic coronary angiography (Fig 2Down). Levels (pmol/mmol creatinine) were 64.7±9.5 before the procedure compared with 84.9±10.6 (P=.02) and 81.4±12.7 (NS) in the first 6 hours and 6 to 12 hours after the procedure, respectively.



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  		Figure 2. Urinary 8-epi-PGF2{alpha} excretion is increased in the first 6 hours (**P<.0l) after angioplasty in patients with acute MI (black bars; n=7) and returns toward baseline by 12 to 24 hours after the procedure. There is a minor increase in 8-epi-PGF2{alpha} levels after diagnostic coronary angiography (white bars; n=11; *P<.05) and elective angioplasty (shaded bars; n=15; *P<.05).

There also was a modest, but significant, increase in urinary 8-epi-PGF2{alpha} after elective coronary PTCA (Fig 2Up). Levels rose from 88.7±7.5 at baseline to 114.3±16.1 (P=.01) in the first 6 hours and 100.3±9.4 (NS) at 6 to 12 hours after successful one-vessel PTCA.

In contrast to controls, urinary 8-epi-PGF2{alpha} levels were markedly elevated after PTCA for acute MI. Levels (pmol/mol creatinine) in the 7 patients requiring PTCA rose from 105±17.8 in the ED to 230±66 (P=.009) in the first 6 hours, 166±31 (NS) at 6 to 12 hours, and 122±18 (NS) at 12 to 24 hours after the procedure (Fig 2Up). Levels were similarly elevated in the overall group (n=10), whether they required PTCA or had been reperfused with thrombolytics alone (before, 87.2±15; 0 to 6 hours, 183±51.2 [P=.004], 6 to 12 hours, 136±26 [P=.05], and 12 to 24 hours, 100±16 [NS]). There was a notable heterogeneity in the increment in urinary 8-epi-PGF2{alpha} among individual patients (Fig 3Down); however, there was an increase in IsoP biosynthesis corresponding to the time of the procedure in all patients, with maximal fold increases over basal ranging from 1.3 to 3.7. Levels had declined to preprocedural values by 12 to 24 hours after the PTCA.



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  		Figure 3. Urinary excretion of 8-epi-PGF2{alpha} (fold increase over basal) in individual patients with acute MI (n=10).

The time course of altered urinary 8-epi-PGF2{alpha} excretion corresponded to the rise in serum creatine phosphokinase release observed in the patients with acute MI (Fig 4Down). The peak in serum creatine phosphokinase in these patients was consistent with early reperfusion.37



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  		Figure 4. There was a similar temporal pattern of urinary 8-epi-PGF2{alpha} excretion ({bullet}) and serum creatine phosphokinase levels ({blacksquare}) in patients after thrombolysis for acute MI.

Urinary IPF2{alpha}-I levels (pmol/mmol creatinine) were higher than 8-epi-PGF2{alpha} levels in patients presenting with acute MI (466±91 versus 105±7.8; P=.003). Similar to 8-epi-PGF2{alpha}, urinary IPF2{alpha}-I also increased 0 to 6 hours after PTCA (833±153; P=.001) and returned toward preprocedure values over time (631±160 [NS] at 6 to 12 hours and 457±102 [NS] at 12 to 24 hours after PTCA). Furthermore, urinary levels of 8-epi-PGF2{alpha} and IPF2{alpha}-I were closely correlated (r=.68; P<.0001) in patients undergoing PTCA for AM1 (Fig 5Down).



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  		Figure 5. Correlation (r=.68; P<.0001; n=33) of urinary 8-epi-PGF2{alpha} and IPF2{alpha}-I levels in patients receiving thrombolytic agents for acute MI.


*    Discussion
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 up arrowAbstract
 up arrowIntroduction
 up arrowMethods
 up arrowResults
 *Discussion
down arrowReferences
 
F2 IsoPs are free radical–catalyzed PGF2{alpha} isomers formed in situ from arachidonic acid esterified in the membrane phospholipid.38 They are cleaved out, presumably by a phospholipase A2, and appear in plasma and urine. Unlike conjugated dienes or lipid hydroperoxides,8,9 they represent chemically stable end products of lipid peroxidation. Recently, isomers of PGE and PGD17 as well as isoleukotrienes39 and isothromboxanes40 have been described. 8-Epi-PGF2{alpha}, an abundant F2 IsoP, has been ascribed biological activity: it has been speculated to ligate a receptor distinct from but closely related to that for thromboxane A2.41,42

Urinary determination of F2 IsoP excretion may be a sensitive index of oxidant stress.15,24,43 Excretion is increased in an animal model of free radical injury,44 in humans after poisoning with acetaminophen and paraquat,15 in patients with alcoholic liver disease.45 and in chronic cigarette smokers,23,43 diverse settings in which free radical–mediated tissue injury is strongly implicated. We also provided preliminary evidence in the last group that administration of vitamin C, which is deficient in smokers,46 depresses urinary 8-epi-PGF2{alpha} excretion. 8-Epi PGF2{alpha} may reflect lipid peroxidation in atherogenesis. For example, zymosan-stimulated human monocytes, coincubated in vitro with LDL, markedly augment 8-epi-PGF2{alpha} formation.47 This increase is prevented by pretreatment of the monocytes with antioxidants. Furthermore, we demonstrated a strong correlation between traditional indices of oxidant stress and 8-epi-PGF2{alpha} generation in LDL particles oxidized by CuCl2 in vitro.24 Consistent with these observations, we detected 8-epi-PGF2{alpha} in human atherosclerotic plaque and immunolocalized it to macrophages and smooth muscle cells within the lesion.48

Recently, we reported the first evidence of increased IsoP generation in syndromes of coronary reperfusion.24 Urinary 8-epi-PGF2{alpha} excretion was increased in a canine model of coronary thrombolysis, in patients with acute MI treated with thrombolytic drugs, and after clamp release in patients undergoing CABG. The increase in urinary 8-epi-PGF2{alpha} during coronary reperfusion corresponded in both time and magnitude to changes in spin-trapping of PBN adducts, as detected by EPR performed ex vivo in patients undergoing CABG.

The predominant pathway for 8-epi-PGF2{alpha} formation in vivo is via free radical–catalyzed transformation of arachidonic acid. However, we have shown that 8-epi-PGF2{alpha}, unlike other F2 IsoPs, is also a minor product of COX-1 in activated human platelets21 and of COX-2 in monocytes in vitro.47 Many of the clinical syndromes in which free radicals have been implicated are associated with platelet activation, including reperfusion with thrombolytic drugs49,50 and PTCA.51 A formal evaluation of the effects of aspirin administration in chronic cigarette smokers, a setting of modest COX-1 activation compared with thrombolysis, suggests that the COX-1 pathway is a trivial contributor to overall 8-epi-PGF2{alpha} formation, as reflected by its excretion in urine.23 Whether this remains true in settings of marked COX activation, such as thrombolysis for acute MI, is currently unclear.

The present study extends and confirms our findings of increased 8-epi-PGF2{alpha} biosynthesis in syndromes of coronary reperfusion.24 It offers the first evidence of oxidative stress in patients undergoing PTCA for acute MI with the use of noninvasive measurement of IsoPs. The temporal relationship between maximal urinary IsoP excretion and PTCA suggests an association between reperfusion and IsoP generation. A less impressive increase was evident in patients undergoing elective PTCA, in whom the degree of ischemia/reperfusion was presumably less pronounced. Of note, diagnostic angiography was associated with a small increment in IsoP biosynthesis. A similar minor increment in urinary prostaglandin has been reported during diagnostic catheterization.51 Previously, we provided evidence consistent with the measurement of 8-epi as a noninvasive index of oxidant stress.15,23,24,52 We now report that a second F2 IsoP, IPF2{alpha}-I, is even more abundant in the urine of patients with coronary artery disease. Importantly, this compound is not formed by COX in vitro or in vivo (J.A.L., P.B. Barry, D. Pratico, J. Rokach, and G.A.F., unpublished data, 1997). Like 8-epi-PGF2{alpha}, there also is a marked increase in excretion of IPF2a-I after PTCA in acute MI patients. The strong correlation between IPF2{alpha}-I and 8-epi-PGF2{alpha} implies that the increment in generation of 8-epi-PGF2{alpha} reflects lipid peroxidation rather than COX activation. This finding is consistent with our previous observations, suggesting that the COX pathway is a trivial contributor to overall 8-epi-PGF2{alpha} generation in vivo.23

The source of IsoPs in urine is likely to be conditional on the experimental setting or the disease under study, as is the case with conventional PGs.53,54 However, we assume that urinary 8-epi-PGF2{alpha} is likely to reflect oxidant stress in tissues other than the kidney. Thus, although studies of the formal disposition of 8-epi-PGF2{alpha} in humans have not been reported, both plasma and urinary 8-epi-PGF2{alpha} are elevated in liver cirrhosis,45 and both plasma and urinary levels of "total" F2-IsoPs are elevated in smokers,43 two syndromes of presumed extrarenal oxidant stress.

A limitation to the study of oxidant injury in cardiovascular disease has been the imprecision of analytical indices of the process in vivo8,9 and the use of ex vivo methodology that is of arguable relevance to what is actually occurring in vivo.11 The absence of appropriate quantitative markers of oxidant stress in vivo has frustrated the ability to develop a rational basis for the dose selection and clinical evaluation of antioxidant drugs and vitamins.55–57 Thus, there is strong evidence for free radical generation and related myocardial dysfunction in animal models of myocardial reperfusion injury using in vivo administration of chemicals that form spin adducts detected by EPR.45 Suppression of such free radical–derived signals with a variety of antioxidants, either alone or in combination, results in attenuation of myocardial dysfunction in these models. The direct EPR techniques used in these studies are not applicable in humans. Therefore, clinical investigators have had to rely on ex vivo spin-trapping techniques and the measurement of less reliable markers of lipid peroxidation, such as malondialdehyde and lipid hydroperoxides. These studies have provided evidence suggestive of free radical generation in humans during PTCA,10,27 acute MI,28,29 and CABG.30,31 However, there has not been a systematic approach to the evaluation of antioxidant strategies in reperfusion syndromes in humans. This has resulted in some confusion. Thus, the lack of clinical efficacy of recombinant human superoxide dismutase given to patients undergoing PTCA for acute MI33 could have been due to a lack of therapeutic effect of this specific agent, the dose selected, or the relative unimportance of oxidant stress in this clinical setting. In contrast, administration of N-acetylcysteine with streptokinase to a group of patients with acute MI34 reportedly suppressed plasma maldionaldehyde, a nonspecific index of oxidant stress, and preserved left ventricular function.

Our findings are of potential importance in clinical practice. First, these noninvasive approaches could be used to determine the relationship between oxidative stress during MI and clinical outcome in larger populations of patients. Our recent development of an immunoassay for 8-epi-PGF2{alpha} would be more applicable than GC/MS to such an application.58 Second, these findings suggest that there is a rational basis for evaluating antioxidants as therapy adjuvant to reperfusion strategies, such as PTCA and thrombolytic drugs. Finally, these noninvasive, quantitative indices of oxidant stress may be used to provide a rational basis for dose selection before evaluation of antioxidant strategies in cardiovascular disease.


*    Selected Abbreviations and Acronyms
 
CABG = coronary artery bypass graft surgery
COX = cyclooxygenase
EPR = electroparamagnetic resonance
GC = gas chromatography
IsoP = isoprostane
MI = myocardial infarction
MS = mass spectrometry
PG = prostaglandin
PTCA = percutaneous transluminal coronary angioplasty
ROS = reactive oxygen species


*    Acknowledgments
 
This study was supported by grants from the Health Research Board of Ireland (Dr Reilly), the Wellcome Trust (Dr FitzGerald and Dr Delanty), and the National Institutes of Health (grant HL-54500) (Dr FitzGerald). Dr FitzGerald is the Robinette Foundation Professor of Cardiovascular Medicine. We are indebted to Marie Mai Lariviere, RN, for her assistance with sample and data collection.


*    Footnotes
 
Reprint requests to Dr G.A. FitzGerald, Center for Experimental Therapeutics, 905 Stellar Chance Building, The University of Pennsylvania, 422 Curie Blvd, Philadelphia, PA 19104.

Presented in part at the annual meeting of the American Heart Association (November 1996) and at the Winter Prostaglandin Meeting (Keystone, January 1997).

Received May 1, 1997; revision received July 22, 1997; accepted August 2, 1997.


*    References
up arrowTop
 up arrowAbstract
 up arrowIntroduction
 up arrowMethods
 up arrowResults
 up arrowDiscussion
 *References
 

  1. Halliwell B. Free radicals, antioxidants, and human disease: curiosity, cause or consequence? Lancet. 1994;344:721–724.[Medline] [Order article via Infotrieve]
  2. Kloner RA. Does reperfusion injury exist in humans? J Am Coll Cardiol. 1993;21:53745.
  3. Bolli R. Myocardial `stunning' in man. Circulation. 1992;86:1671–1691.[Medline] [Order article via Infotrieve]
  4. Sekili S, McCay PB, Li X-Y, Zughaib M, Sun J-Z, Tang L, Thornby JI, Bolli R. Direct evidence that the hydroxyl radical plays a pathogenetic role in myocardial `stunning' in the conscious dog and demonstration that stunning can be markedly attenuated without subsequent adverse effects. Circ Res. 1993;73:705–723.[Abstract]
  5. Li X-Y, McCay PB, Zughaib M, Jeroudi MO, Triana F, Bolli R. Demonstration of free radical generation in the `stunned' myocardium in the conscious dog and identification of major differences between conscious and open-chest dogs. J Clin Invest. 1993;92:1025–1041.
  6. Halliwell B, Chirico S. Lipid peroxidation: its mechanism, measurement and significance. Am J Clin Nutr. 1993;57(suppl):715S–725S.
  7. Holley AK, Cheeseman KH. Measuring free radical reactions in vivo. Br Med Bull. 1992;49:494–505.[Abstract/Free Full Text]
  8. Wade CR, van Rij AM. Plasma thiobarbituric acid reactivity: reaction conditions and the role of iron, antioxidants and lipid peroxyradicals on the quantitation of plasma lipid peroxides. Life Sci. 1988;43:1085–1093.[Medline] [Order article via Infotrieve]
  9. Yamamoto Y, Frei B, Ames B. Assay of lipid hydroperoxides using high performance liquid chromatography with isoluminol chemiluminesence detection. Methods Enzymol. 1990;186:371–80.[Medline] [Order article via Infotrieve]
  10. Grech ED, Dodd H1F, Bellamy CM, Perry RA, Morrison WL, Ramsdale DR. Free radical generation during angioplasty reperfusion for acute myocardial infarction. Lancet. 1993;341:990–991.[Medline] [Order article via Infotrieve]
  11. Esterbauer H, Stnegl G, Puhl H, Rotheneder M. Continuous monitoring of in vitro oxidation of human low density Lipoprotein. Free Rad Res Commun. 1989;6:67–75.[Medline] [Order article via Infotrieve]
  12. Loft S, Astrup A, Buemann B, Poulsen HE. Oxidative DNA damage correlates with oxygen consumption in humans. FASEB J. 1994;8:534–537.[Abstract]
  13. Leeuwenburgh C, Rasmussen JE, Hsu FF, Mueller DM, Pennathur S, Heinecke JW. Mass spectrometric quantification of markers for protein oxidation by tyrosyl radical, copper and hydroxyl radical in low density lipoprotein isolated from human atherosclerotic plaques. J Biol Chem. 1997;272:3520–3526.[Abstract/Free Full Text]
  14. Morrow JD, Roberts LJ II. The isoprostanes: current knowledge and directions for future research. Biochem Pharm. 1996;51:1–9.[Medline] [Order article via Infotrieve]
  15. Delanty N, Reilly M, Pratico D, FitzGerald DJ, FitzGerald GA. 8-Epi PGF2{alpha}: specific analysis of an isoeicosanoid as an index of oxidant stress in vivo. Br J Clin Pharm. 1996;42:15–19.[Medline] [Order article via Infotrieve]
  16. Morrow JD, Harris TM, Roberts LJ II. Noncyclooxygenase oxidative formation of a series of novel prostaglandins: analytical ramifications for measurement of eicosanoids. Anal Biochem. 1990;184:1–10.[Medline] [Order article via Infotrieve]
  17. Morrow JD, Minton TA, Mukundan CR, Campbell MD, Zackert WE, Daniel VC, Badr KF, Blair IA, Roberts LJ II. Free radical-induced generation of isoprostanes in vivo: evidence for the formation of D-ring and E-ring isoprostanes. J Biol Chem. 1994;269:4317–4326.[Abstract/Free Full Text]
  18. Waugh RJ, Murphy RC. Mass spectrometric analysis of four regioisomers of F2 isoprostanes formed by free radical oxidation of arachidonic acid. J Am Soc Mass Spectrom. 1996;7:490–499.
  19. Takahashi K, Nammour TM, Fukunaga M, Ebert J, Morrow JD, Roberts II LJ, Hoover RL, Badr KF. Glomerular actions of a free radical generated novel prostaglandin, 8-epi-prostaglandin F2{alpha} in the rat: evidence for interaction with thromboxane A2 receptors. J Clin Invest l992;90:136–141.
  20. Fukunaga M, Makita N, Roberts LJ, II, Morrow JD, Takahashi K, Badr KF. Evidence for the existence of F2-isoprostane receptors on rat vascular smooth muscle cells. Am J Physiol l993;264:1619–1624.
  21. Pratico D, Lawson JA, FitzGerald GA. Cyclooxygenase-dependent formation of the isoprostane, 8-epi prostaglandin F2{alpha}. J Biol Chem. 1995;270:9800–9808.[Abstract/Free Full Text]
  22. Adiyaman M, Lawson JA, Hwang S-W, Khanapure SP, FitzGerald GA, Rokach J. Total synthesis of a novel isoprostane IPF2{alpha}-I and its identification in biological fluids. Tetrahed Lett. 1996;37:4849–4852.
  23. Reilly MP, Delanty N, Lawson JA, FitzGerald GA. Modulation of oxidant stress in vivo in chronic cigarette smokers. Circulation. 1996;94:19–25.[Medline] [Order article via Infotrieve]
  24. Delanty N, Reilly M, Lawson JA, McCarthy J, Wood F, FitzGerald DJ, FitzGerald GA. 8-Epi PGF2{alpha} generation during coronary reperfusion: a potential quantitative marker of oxidant stress in vivo. Circulation. 1997;95:2492–2499.[Medline] [Order article via Infotrieve]
  25. Galli M, Marcassa C, Bolli R, Gianuzzi P, Temporelli PL, Imparato A, Silva Oreggo PL, Giubbini R, Giordano A, Tavazzi L. Spontaneous delayed recovery of reperfusion and contraction after the first 5 weeks after anterior infarction. Circulation. 1994;90:1386–1397.[Medline] [Order article via Infotrieve]
  26. Breisblatt WM, Stein KL, Wolfe CJ, Follansbee WP, Capozi J, Armitage JM, Hardesty RL. Acute myocardial dysfunction and recovery: a common occurrence after coronary bypass surgery. J Am Coll Cardiol. 1990;15:1261–1269.[Abstract]
  27. Coghlan JG, Flitters WD, Holley AK, Norell M, Mitchell AG, Isley CD, Slater TF. Detection of free radicals and cholesterol hydroperoxides in blood taken from the coronary sinus of man during percutaneous transluminal coronary angioplasty. Free Rad Res Commun. 1991;14:409–417.[Medline] [Order article via Infotrieve]
  28. Davies SW, Ranjadayalan K, Wickens DC, Dorinandy TL, Tinunis AD. Lipid peroxidation associated with successful thrombolysis. Lancet. 1990;335:741–743.[Medline] [Order article via Infotrieve]
  29. Purvis J, Young L Lightbody J, Trimble E, Adgey J. Free radical production following thrombolytic therapy in acute myocardial infarction. Circulation. 1992;86(suppl I):I-804. Abstract.
  30. Coghlan JG, Flitter WD, Clutton SM, Panda R, Daly R, Wright G, Isley CD, Slater TF. Allopurinol pretreatment improves postoperative recovery and reduces lipid peroxidation in patients undergoing coronary bypass grafting. J Thorac Cardiovasc Surg. 1994;107:248–256.[Abstract/Free Full Text]
  31. Tortolani AJ, Powell SR, Misik V, Weglicki WB, Pogo GJ, Kramer JH. Detection of alkoxyl and carbon-centered free radicals in coronary sinus blood from patients undergoing elective cardioplegia. Free Rad Biol Med. 1993;14:421–426.[Medline] [Order article via Infotrieve]
  32. Ferrari R, Alfieri O, Curello S, Ceconi C, Cargnoni A, Marzollo P, Pardinni A, Caradonna E, Visioli O. Occurrence of oxidative stress during reperfusion of the human heart. Circulation. 1990;81:201–211.[Medline] [Order article via Infotrieve]
  33. Flaherty JT, Pitt B, Gruber JW, Heuser RR, Rothbaum DA, Burwell LR, George BS, Kereiakes DJ, Deitchrnan D, Gustafson N, Brinker JA, Becker LC, Mancini J, Topol E, Werns SW. Recombinant human superoxide dismutase (h-SOD) fails to improve recovery of ventricular function in patients undergoing coronary angioplasty for acute myocardial infarction. Circulation. 1994;89:1982–1991.[Medline] [Order article via Infotrieve]
  34. Arstall MA, Yang J, Stafford 1, Betts WH, Horowitz JD. N-Acetylcysteine in combination with nitroglycerin and streptokinase for the acute myocardial infarction. Circulation. 1995;92:2855–2862.[Medline] [Order article via Infotrieve]
  35. Ohman EM, Slovak IP, Anderson RL, Grossman WJ, Barbeau GR, Butler JF, Frey MT, Talley DJ, Leimberger 1D, Scherer JC, Kleiman NS. Potent inhibition of thrombin with Efegatran in combination with t-PA in acute myocardial infarction: results of a multicenter randomized dose ranging trial. Circulation. 1996;94(suppl I):I-430. Abstract.
  36. Llobat-Estelles M, Sevillano-Cabeja A, Campins-Falco P. Kinetic and chemometric studies of the determination of creatinine using the Jaffe reaction, part I: kinetics of the reaction: analytical conclusion. Analyst. 1989;114:597–602.[Medline] [Order article via Infotrieve]
  37. Karajounis L, Moreno F, Menlove RL, Ipsen S, Anderson JL. Effect of early thrombolytic therapy (anistrepase versus streptokinase) on enzymatic and electrocardiographic infarct size in acute myocardial infarction: TEAM-2 Investigators. Am J Cardiol. 1991;68:848–856.[Medline] [Order article via Infotrieve]
  38. Morrow JD, Awad JA, Boss HJ, Blair IA, Roberts LJ II. Non-cyclooxygenase derived prostanoids (F2-isoprostanes) are formed in situ on phospholipids. Proc Natl Acad Sci U S A.. 1992;89:10721–10725.[Abstract/Free Full Text]
  39. Harrison KA, Murphy RC. Isoleukotrienes: biologically active free radical products of lipid peroxidation. J Biol Chem. 1995;270:17273–17278.[Abstract/Free Full Text]
  40. Pratico D, Lawson J, FitzGerald GA. Isothromboxane formation by human monocytes. Blood. 1995;86:509. Abstract.
  41. Yin K, Halushka PV, Yan Y, Wong PY. Antiaggregatory activity of 8-epi prostaglandin F2a and other F-series prostanoids and their binding to thromboxane A2/prostaglandin H2 receptors in human platelets. J Pharmacol Exp Ther. 1994;270:1192–1196.[Abstract/Free Full Text]
  42. Pratico D, Smyth EM, Violi F, FitzGerald GA. Local amplification of platelet function by 8-epi PGF2{alpha} is not mediated by thromboxane receptor isoforms. J Biol Chem. 1996;271:14916–14924.[Abstract/Free Full Text]
  43. Morrow JD, Frei B, Atkinson AW, Gaziano M, Lynch SM, Shyr Y, Strauss WE, Oates JA, Roberts LJ II. Increase in circulating products of lipid peroxidation (F2 isoprostanes) in smokers: smoking as a cause of oxidative damage. N Engl J Med. 1995;332:1198–1203.[Abstract/Free Full Text]
  44. Morrow JD, Awad JA, Kato T, Takahashi K, Badr KF, Roberts LJ II, Burk RF. Formation of novel non–cyclooxygenase-derived prostanoids (F2-isoprostane) in carbon tetrachloride hepatotoxicity: an animal model of lipid peroxidation. J Clin Invest. 1992;90:2502–2507.
  45. Pratico D, Basili B, Cordova C, Iuliano L, Ferro D, Camastra C, FitzGerald GA, Violi F. Increased oxidant stress in liver cirrhosis. J Invest Med. 1996;44:301A. Abstract.
  46. Schectman G, Byrd JC, Gruchow HW. The influence of smoking on vitamin C status in adults. Am J Public Health. 1989;79:158–162.[Abstract/Free Full Text]
  47. Pratico D, FitzGerald GA. Generation of 8-epi prostaglandin F2{alpha} by human monocytes. J Biol Chem. 1996:271:8919–8924.
  48. Pratico D, Iuliano L, Mauriello A, Spagnoli L, Lawson JA, Maclouf J, Violi F, Fitzgerald GA. Localization of distinct F2-isoprostanes in human atherosclerotic lesions. J Clin Invest. 1997;100:2028-2034.
  49. FitzGerald DJ, Catella F, Roy L, Fitzgerald GA. Platelet activation in vivo after intravenous streptokinase in patients with acute myocardial infarction. Circulation. 1988;77:142–150.[Medline] [Order article via Infotrieve]
  50. Kerins DM, Roy L, FitzGerald GA, Fitzgerald DJ. Platelet and vascular function during coronary thrombolysis with tissue-type plasminogen activator. Circulation. 1989;80:1718–1725.[Medline] [Order article via Infotrieve]
  51. Braden G, Knapp HR, FitzGerald GA. Suppression of eicosanoid formation during coronary angioplasty by fish oil and aspirin. Circulation. 1991;83:679–685.
  52. Reilly MP, Delanty N, Tremoli E, Rader D, FitzGerald GA. Elevated levels of 8-epi in familial hypercholesterolemia: evidence of oxidative stress in vivo. Circulation. 1996;94(suppl I):I-683. Abstract.
  53. FitzGerald GA, Pedersen AC, Patrono C. Analysis of prostacyclin and thromboxane biosynthesis in cardiovascular disease. Circulation. 1983;67:1174–1177.[Medline] [Order article via Infotrieve]
  54. Patrono C, Ciabattoni G, Pugliesse F, Pierucci A, Blair IA, FitzGerald GA. Estimated rate of thromboxane secretion into the circulation of normal man. J Clin Invest. 1986;77:590–594.
  55. Hennekens CH, Buring JE, Peto R. Antioxidant vitamins: benefits not yet proved. N Engl J Med. 1994;330:1080–1081.[Free Full Text]
  56. Hennekens CH, Buring JE, Manson J, Stampfer M, Rosner B, Cook NR, Belanger C, LaMotte F, Gaziano JM, Ridker PM, Willet W, Peto R. Lack of effect of long-term supplementation with beta carotene on the incidence of malignant neoplasms and cardiovascular disease. N Engl J Med. 1996;334:1145–1149.[Abstract/Free Full Text]
  57. Stephens NG, Parsons PM, Kelly F, Cheeseman K, Mitchinson MJ, Brown MJ. Randomized controlled trial of vitamin E in patients with coronary disease: Cambridge Heart Antioxidant Study. Lancet. 1996;347:781–786.[Medline] [Order article via Infotrieve]
  58. Wang Z, Ciabattoni G, Creminon C, Lawson JA, FitzGerald GA, Patrono C, Maclouf J. Immunoassay of urinary 8-epi PGF2{alpha}. J Pharmacol Exp Ther. 1995;275:94–100.[Abstract/Free Full Text]



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