Peroxides, Peroxyl, and Hydroxyl Radicals

Peroxide, superoxide, peroxyl and hydroxyl radicals belong to this group of active oxygen species. Hydroxyl radicals, the most reactive species, are generated from the reaction between peroxide and superoxide . 

Several powerful oxidants are produced during the course of metabolism, in both blood cells and most other cells of the body. These include superoxide (02 ), hydrogen peroxide (H2O2), peroxyl radicals (ROO ), and hydroxyl radicals (OH ). The last is a particularly reactive molecule and can react with proteins, nucleic acids, lipids, and other molecules to alter their structure and produce tissue damage. The reactions listed in Table 52-4 play an important role in forming these oxidants and in disposing of them each of these reactions will now be considered in turn. 

The evolution of fluoride ions in actual fuel cell effluent and during laboratory accelerated life studies has been reported. One common example of radical generation from peroxide decomposition is in the Fenton test, where peroxyl or hydroxyl radicals can be formed through the reaction of hydrogen peroxide with Fe(II) (Scheme 3.2). 

The lung also possesses nonenzymatic antioxidants such as vitamin E, beta-carotene, vitamin C, and uric acid. Vitamin E is lipid-soluble and partitions into lipid membranes, where it is positioned optimally for maximal antioxidant effectiveness. Vitamin E converts superoxide anion, hydroxyl radical, and lipid peroxyl radicals to less reactive oxygen metabolites. Beta-carotene also accumulates in cell membranes and is a metabolic precursor to vitamin A. Furthermore, it can scavenge superoxide anion and react directly with peroxyl-free radicals, thereby serving as an additional lipid-soluble antioxidant. Vitamin C is widely available in both extracellular and intracellular spaces where it can participate in redox reactions. Vitamin C can directly scavenge superoxide and hydroxyl radical. Uric acid formed by the catabolism of purines also has antioxidant properties and primarily scavenges hydroxyl radical and peroxyl radicals from lipid peroxidation. 

Radicals generated during peroxidation of lipids and proteins show reactivity similar to that of the hydroxyl radical however, their oxidative potentials are lower. It is assumed that the reactive alkoxyl radicals rather than the peroxyl radicals play a part in protein fragmentation secondary to lipid peroxidation process, or protein exposure to organic hydroperoxides (DIO). Reaction of lipid radicals produces protein-lipid covalent bonds and dityrosyl cross-links. Such cross-links were, for example, found in dimerization of Ca2+-ATPase from skeletal muscle sarcoplasmic reticulum. The reaction was carried out in vitro by treatment of sarcoplasmic reticulum membranes with an azo-initiator, 2,2/-azobis(2-amidinopropane) dihydrochloride (AAPH), which generated peroxyl and alkoxyl radicals (V9). 

Low molecular weight antioxidants react with ROS in cell compartments which for some reasons are lack of antioxidant enzymes. Thus, suppression of bifurcate chain reactions of lipid peroxidation in hydrophobic core of cell membrane is mostly effectively performed by vitamin E (a-tocopherol). Interaction of lipid molecules with hydroxyl radical in the absence of vitamin E results in bifurcation of oxidative processes and formation of peroxyl and alcoxyl radicals. They are quickly accumulated in the restricted volume of the membrane and reaction began to be uncontrolled. a-Tocopherol interacts with peroxyl radicals with high affinity, reduces them and is then oxidized itself into relatively nonactive phenoxyl radical [8]. The latter can be accumulated within the bilayer until it will be returned to initial state by reduction by ascorbate [9]. Pair Vitamin E - Vitamin C is a good example of a mutual interaction between hydrophobic and hydrophilic low molecular weight antioxidants. Recently, tight relations were demonstrated for several natural antioxidants which interaction balances the red/ox state of the cell [3.5.10-12]. Figure 4 demonstrates such interaction between some of them. 

Free radicals produced in vivo include superoxide, the hydroxyl radical, nitric oxide, oxygen-centered organic radicals such as peroxyl and alkoxyl radicals, and sulfur-centered thiyl radicals. Other oxygen-containing reactive species that are not radicals are also formed. These include hydrogen peroxide, peroxynitrite, and hypochlorous acid. While these are not radical species, they are actually or potentially damaging oxidants. The collective term ROS is often used to describe both radical and nonradical species. 

As strong antioxidants and scavengers of superoxide, hydroxyl and peroxyl radicals, tea flavonoids can suppress radical chain reactions and terminate lipid peroxidation (Kumamoto and Sonda, 1998, Yang and Wang, 1993). 

Ascorbate is known to act as a water-soluble antioxidant, reacting rapidly with superoxide, hydroxyl and peroxyl radicals. However, reduced ascorbate can react non-enzymatically with molecular oxygen to produce dehydroascorbate and hydrogen peroxide. Also, ascorbate in the presence of light, hydrogen peroxide and riboflavin, or transition metals (e.g. Fe, Cu " ), can give rise to hydroxyl radicals (Delaye and Tardieu, 1983 Ueno et al., 1987). These phenomena may also be important in oxidative damage to the lens and subsequent cataract formation. 

In Scheme 7, the peroxidic 0-0 bond of the hydroperoxyl group is broken together with /1-scission of the formed alkoxyl radical, and, further, ring closure of alkyl peroxyl diradical may occur. The process generates a hydroxyl radical, methylcarbonyl terminal groups (-CH2-CO-CH3) and dioxetane. The latter is unstable and decomposes into an excited triplet state of formaldehyde and/or excited triplet state of methylcarbonyls (Scheme 8). 

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