Original Contributions
The red wine antioxidant resveratrol protects isolated rat hearts from ischemia reperfusion injury

https://doi.org/10.1016/S0891-5849(99)00063-5Get rights and content

Abstract

The consumption of red wine has been reported to impart a greater benefit in the prevention of coronary heart disease than the consumption of other alcoholic beverages. This beneficial effect is increasingly being attributed to certain antioxidants comprising the polyphenol fraction of red wine such as transresveratrol. In the present study, we investigated the potential cardioprotective effects of resveratrol in the face of ischemia reperfusion (I/R) injury. Isolated perfused working rat hearts after stabilization were perfused with Krebs-Henseleit Bicarbonate buffer (KHB) either in the presence or absence of transresveratrol (RVT) at a concentration of 10 μM for 15 min prior to subjecting them to 30 min of global ischemia followed by 2 h of reperfusion. Left ventricular functions were monitored at various timepoints throughout the reperfusion period to assess the extent of postischemic recovery in comparison with baseline values. Coronary perfusate samples were also collected to determine malonaldehyde (MDA) levels. The results demonstrated that RVT exhibited significant myocardial protection. This was evidenced by improved recovery of post-ischemic ventricular function including developed pressure and aortic flow as compared to the control group (KHB). Values for developed pressure in the RVT-treated group were significantly higher than those in the control group throughout the reperfusion period (71.09 ± 4.88 mmHg vs. 58.47 ±3.88 mmHg, 68.87 ± 5.07 mmHg vs. 49.74 ± 2.65 mmHg and 51.67 ± 3.95 mmHg vs. 30.50 ± 4.80 mmHg at reperfusion timepoints R-15, R-60, and R-120, respectively). From R-30 onwards, aortic flow was markedly higher in the RVT treated group as compared with the control group, the differences being most significant at R-90 (32.45 ± 2.19 ml/min vs. 19.83 ± 1.62 ml/min) and R-120 (27.15 ±2.27 ml/min vs. 14.10 ± 1.69 ml/min). In contrast to the KHB treated group, the RVT-treated group displayed significant reduction in MDA formation especially in the immediate early reperfusion period (63.71 ± 8.19 pM/ml vs. 130.86 ± 4.76 pM/ml, 63.84 ± 15.62 pM/ml vs. 156.99 ± 18.93 pM/ml, 71.29 ± 2.80 pM/ml vs. 129.5 ± 10.30 pM/ml and 56.25 ± 5.79 pM/ml vs. 127.99 ±3.50 pM/ml at timepoints R-1, R-3, R-5, and R-7, respectively) indicating a reduction in I/R injury related oxidative stress. Infarct size was markedly reduced in the RVT group when compared with the control group (10.57 ± 0.35% vs. 36.27 ± 5.28%). In vitro studies revealed RVT to be a potent scavenger of peroxyl radicals suggestive of a probable mechanism involved in the protective ability of RVT. The results of this study indicate that resveratrol possesses cardioprotective effects which may be attributed to its peroxyl radical scavenging activity.

Introduction

Resveratrol (3,5,4′-trihydroxystilbene) is a natural phytoalexin found in a wide variety of plant species including grapes. It is abundantly present in the seeds and skin of grapes and constitutes one of the major components of red wine [1]. Resveratrol has been found to exert several interesting biologic effects. In addition to its anti-inflammatory activity attributed to cycloxygenase-1 (COX-1) inhibition (ED50 = 15 μM) [2], it was also found to inhibit ribonucleotide reductase at a concentration of 100 μM [3] and inhibit cycloxygenase-2 (COX-2) transcription and activity (IC50 = 32.2 μM) [4]. The latter two have been proposed to be possible mechanisms for its cancer chemopreventive activity. Recently, resveratrol was found to prevent lipid peroxidation at a concentration of 1.5 μM [5] and lipid peroxidation-induced cell death at a concentration of 10 μM [6]. Resveratrol was also found to stimulate nitric oxide (NO) production in the endothelial cell [7] and both direct and indirect vasodilatory effects on the blood vessels by NO-dependent (> 30 μM) and NO-independent (> 60 μM) mechanisms were demonstrated, respectively.

A significant amount of evidence is present in the literature to support the role of oxygen free radicals in the pathogenesis of myocardial ischemia reperfusion injury [8]. The presence of free radicals in the ischemic reperfused myocardium has been confirmed both directly and indirectly [9]. This receives further support from the evidence that a variety of free radical scavengers and antioxidants are capable of ameliorating ischemia reperfusion injury [10]. Among the oxygen free radicals the most detrimental effects to cells are caused by the hydroxyl radical (OHradical dot). Although organic peroxyl radicals possess a relatively lower oxidising ability in comparison to the hydroxyl radicals, they are however capable of exerting considerable damage to tissues in biological systems [11]. Furthermore, peroxyl radicals are formed in vivo in membranes and lipoproteins as intermediate products of lipid peroxidation, which is associated with ischemia reperfusion injury. In addition to the above, we and others demonstrated that nitric oxide plays a crucial role in myocardial preservation during ischemic arrest [12]. Enhancement of NO production was associated with myocardial preservation during ischemic arrest while inhibition of NO synthesis enhanced myocardial ischemia reperfusion injury.

The antioxidant property of resveratrol as also its capacity to stimulate endothelial NO production prompted us to investigate whether it is capable of exerting any protective effects in the face of ischemia reperfusion (I/R) injury. So far studies have concentrated on considering the polyphenol fraction of red wine from the point of view of being responsible for the beneficial effects of red wine in the prevention of atherosclerosis and coronary heart disease. However it has never been considered that they may possibly be utilized as a therapeutic or treatment modality in acute scenarios such as I/R injury. The aim of this study was to evaluate the possible role of resveratrol in protecting the myocardium from the damaging effects of I/R injury using the isolated perfused working rat heart.

Section snippets

Materials and methods

All animals used in this study received humane care in compliance with the principles of laboratory animal care formulated by the National Society for Medical Research and Guide for the Care and Use of Laboratory Animals prepared by the National Academy of Sciences and published by the National Institutes of Health (publication no. NIH 85-23, revised 1985). Eighteen male Sprague-Dawley rats (275–300 g) were used in this study. They were provided with food and water ad libitum up until the start

Functional recovery

There were no differences in baseline function between groups A and B. As was expected, on reperfusion, the absolute values of all functional parameters were decreased in both groups as compared with the respective baseline values except in the case of aortic diastolic pressure whose values were observed to undergo an increase. Group B (transresveratrol, 10 μM) displayed significant recovery of postischemic myocardial function. Before using trans-resveratrol at a concentration of 10 μM,

Discussion

In this report we provide evidence that resveratrol possesses potent cardioprotective properties. Resveratrol-treated hearts were resistant to ischemia reperfusion injury as evidenced by improved postischemic ventricular functions and reduced infarct size. In concert MDA formation in the coronary effluents of the resveratrol-treated hearts was significantly lower as compared to those of control animals suggesting reduced free radical formation in the hearts pretreated with resveratrol. In vitro

Acknowledgements

This study was supported in part by National Institutes of Health HL 22559, HL 33889, and HL 34360.

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