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Free Radical Biology of the Cardiovascular System

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Free Radical Biology of the Cardiovascular System Clinical Science (2012) 123, 73–91 (Printed in Great Britain) doi:10.1042/CS20110562 73 REVIEW Free radical biology of the cardiovascular system Alex F. CHEN∗, Dan-Dan CHEN∗, Andreas DAIBER†, Frank M. FARACI‡, Huige LI§, Christopher M. REMBOLD and Ismail LAHER¶ ∗Department of Surgery, McGowan Institute of Regenerative Medicine, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, U.S.A., †2nd Medical Clinic, Molecular Cardiology, Medical Center of the Johannes Gutenberg University, Mainz, Germany, ‡Departments of Internal Medicine and Pharmacology, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, U.S.A., §Department of Pharmacology, University Medical Center, Johannes Gutenberg University, Mainz, Germany, Cardiovascular Division, Department of Internal Medicine, University of Virginia, Charlottesville, VA 22908, U.S.A., and ¶Department of Pharmacology and Therapeutics, Faculty of Medicine, University of British Columbia, 2176 Health Sciences Mall, Vancouver, BC, Canada V6T 1Z3 ABSTRACT Most cardiovascular diseases (CVDs), as well as age-related cardiovascular alterations, are accompanied by increases in oxidative stress, usually due to increased generation and/or decreased metabolism of ROS (reactive oxygen species; for example superoxide radicals) and RNS (reactive nitrogen species; for example peroxynitrite). The superoxide anion is generated by several enzymatic reactions, including a variety of NADPH oxidases and uncoupled eNOS (endothelial NO synthase). To relieve the burden caused by this generation of free radicals, which also occurs as part of normal physiological processes, such as mitochondrial respiratory chain activity, mammalian systems have developed endogenous antioxidant enzymes. There is an increased usage of exogenous antioxidants such as vitamins C and E by many patients and the general public, ostensibly in an attempt to supplement intrinsic antioxidant activity. Unfortunately, the results of large-scale trails do not generate much enthusiasm for the continued use of antioxidants to mitigate free-radical-induced changes in the cardiovascular system. In the present paper, we review the clinical use of antioxidants by providing the rationale for their use and describe the outcomes of several large-scale trails that largely display negative outcomes. We also describe the emerging understanding of the detailed regulation of superoxide generation by an uncoupled eNOS and efforts to reverse eNOS uncoupling. SIRT1 (sirtuin 1), which regulates the expression and activity of multiple pro- and anti-oxidant enzymes, could be considered a candidate molecule for a ‘molecular switch’. − Key words: cardiovascular disease, endothelial progenitor cell, free radical, oxidative stress, sirtuin, superoxide anion (O2 ). Abbreviations: ACE, angiotensin-converting enzyme; ADMA, asymmetric ω-NG,NG-dimethylarginine; AGE, advanced glycation end-product; AngII, angiotensin II; ApoE, apolipoprotein E; ARB, angiotensin receptor blocker; AT1 receptor, AngII type 1 receptor; BH4, tetrahydrobiopterin; CAD, coronary artery disease; CVD, cardiovascular disease; EPC, endothelial progenitor cell; FOXO, forkhead box O; GCH, GTP cyclohydrolase; GPx, glutathione peroxidase; GTN, nitroglycerine; KO, knockout; LDL, low- density lipoprotein; l-NAME, NG-nitro-l-arginine methyl ester; MI, myocardial infarction; mitoQ, mitoquinone; MMP, matrix metalloproteinase; mPTP, mitochondrial permeability transition pore; mtKATP channel, mitochondrial ATP-sensitive potassium − − channel; NOS, NO synthase; eNOS, endothelial NOS; O2 , superoxide anion; ONOO , peroxynitrite; PARP, poly(ADP- ribose) polymerase; PETN, pentaerithrityl tetranitrate; PKC, protein kinase C; PPAR, peroxisome- proliferator-activated receptor; RAS, renin–angiotensin system; RNS, reactive nitrogen species; RONS, reactive oxygen and nitrogen species; ROS, reactive oxygen species; siRNA, small interfering RNA; SIRT1, sirtuin 1; SOD, superoxide dismutase; Cu/Zn-SOD, copper/zinc SOD; Mn-SOD, manganese SOD; UAG, unacylated ghrelin; VEGF, vascular endothelial growth factor. Correspondence: Dr Ismail Laher (email [email protected]). C The Authors Journal compilation C 2012 Biochemical Society 74 A. F. Chen and others INTRODUCTION Cu/Zn-SOD (copper/zinc SOD; SOD1), Mn-SOD (manganese SOD; SOD2) and ec-SOD (extracellular There is much interest in the role of free radical oxidative SOD; SOD3). GPx proteins consume reduced gluta- damage in human diseases. The first description of free thione to convert H2O2 into water and lipid peroxides radicals was by Gomberg in 1900 at the University to their respective alcohols, with GPx1 being the most of Michigan; the term free radicals refers to atoms, abundant selenoperoxidase and a key antioxidant enzyme molecules or ions with unpaired electrons in their in many cell types. Catalase is an intracellular antioxidant outer shells, making them highly reactive in biological enzyme that is mainly located in cellular peroxisomes and systems. This unstable configuration provides energy that to some extent in the cytosol, which catalyses the reaction is released through reactions with adjacent molecules of H2O2 into water and molecular oxygen [9]. Non- such as proteins, lipids, carbohydrates and nucleic acids. enzymatic antioxidants include vitamins E (tocopherol) As a result, oxidative stress is implicated in several and C (ascorbic acid), thiol antioxidants (glutathione, human diseases, including hypertension, atherosclerosis thioredoxin and lipoic acid), melatonin, carotenoids, and vascular diabetic complications, as well as in the natural flavonoids and other compounds [10]. aging process [1]. The majority of free radicals that The growing interest in the possible beneficial roles damage biological systems are oxygen free radicals. of antioxidant supplements in the treatment of CVD Oxygen free radicals or, more generally, ROS (reactive (cardiovascular disease) has contributed to a debate about oxygen species), as well as RNS (reactive nitrogen their value in providing complementary therapies aimed species), are products of normal cellular metabolism at improving standard therapy. There are numerous [2]. The most common RONS (reactive oxygen and explanations for the failure to observe noticeable benefits − nitrogen species) include O2 (superoxide anions), H2O2 with the use of currently available antioxidants. It is • (hydrogen peroxide), OH (hydroxyl radicals), carbon- possible that the agents examined are ineffective and • centred peroxides and peroxyl radicals, NO (NO non-specific, and that the dosing regimens and the • − radicals), NO2 (nitrogen dioxide radicals), ONOO duration of therapy are insufficient. Vitamins C and E − (peroxynitrite), ClO (hypochlorite) and others [3–5]. may have pro-oxidant properties with harmful and − O2 can be formed from different sources such as deleterious interactions. It is also possible that orally xanthine oxidase, NADPH oxidases, uncoupled NOSs administered antioxidants may be inaccessible to the (NO synthases), enzymes of the mitochondrial respir- source of free radicals, particularly if ROS are generated in atory chain and cytochrome P450 mono-oxygenases [1]. intracellular compartments and organelles. Furthermore, Among these enzymes, NADPH oxidases and uncoupled antioxidant vitamins do not scavenge H2O2, which may − eNOS (endothelial NOS) are crucial ROS sources under be more important than O2 in CVD [11]. In the present pathological conditions [6]. Oxidative stress defines a review, we will discuss strategies for the design of newer state with either increased (uncontrolled) formation of antioxidants. For example, it may be more promising to RONS and/or impaired cellular antioxidant defence develop antioxidants that accumulate at the site of oxidant system (e.g. down-regulation of important antioxidant formation (e.g. mitochondrial-targeted antioxidants) or proteins) with subsequent depletion of low-molecular- to inhibit the sources of oxidants (e.g. NADPH oxidase mass antioxidants and a shift in the cellular redox balance inhibitors). Another strategy would be the synthetic [7]. production of intrinsic antioxidant enzymes (e.g. SOD- Organisms have evolved a number of antioxidant mimetic antioxidants). enzymes to combat the burden of increased oxidative stress. An antioxidant refers to any molecule capable of stabilizing or deactivating free radicals before they OXIDATIVE STRESS IN VASCULAR AGING attach to cells. Humans have evolved highly complex AND DISEASE antioxidant systems (enzymatic and non-enzymatic) that work synergistically, and in combination with each Aging produces complex changes in both structure and other, to protect cells and organ systems of the body function throughout the vascular tree [12]. Mechanisms against free radical damage. Antioxidants can be either that underlie vascular homoeostasis and maintain oxygen endogenously produced substances or obtained from delivery often become impaired with aging. For example, exogenous sources, e.g. as a part of a diet or as dietary resting blood flow and most vasodilator responses decline supplements. Some dietary compounds that do not with age. Complex responses including neurovascular neutralize free radicals, but enhance endogenous activity, coupling and endothelium-dependent regulation of can also be classified as antioxidants [8]. vascular tone exhibit age-related dysfunction in both The most efficient enzymatic antioxidants involve GPx experimental animals and humans [13]. Vascular or (glutathione peroxidase), catalase
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