Oxidized LDL

Corona.iy Hea.rt Disea.se, A theory for atherogenesis (120) has been developed whereby oxidation of low density Hpoprotein (LDL) within the arterial wall is the critical first step. It has been hypothesized that sufficient intake of antioxidants would prevent oxidation of LDL and reduce development of coronary heart disease (122). Interest in determining the role of antioxidants in blocking LDL oxidation has led to the development of in vitro test systems. 

Table 7. Antioxidant Potency of Vitamins and Phenolics Based on LDL Oxidation In Vitrei...

Heinecke JW (2001) Is the emperor wearing clothes Clinical trials of vitamin E and the LDL oxidation hypothesis. Arterioscler Thromb Vase Biol 21 1261—1264... 

CHOPRA M, MCLOONE u L, o neill m, WILLIAMS N and THURNHAM DI (1996) Fruit and vegetable supplementation - effect on ex vivo LDL oxidation in hiunans , in Kumpulainen, J T and Saonen, J T (eds), Natural Antioxidants and Food Quality in Atherosclerosis and Cancer Prevention, Cambridge, Royal Society of Chemistry, 150-55. 

KAPIOTIS S, HERMAN M, HELD I, SEELOS C, EHRINGER H, GMEINER B M, (1997) Genistein, the dietary-derived angiogenesis inhibitor, prevents LDL oxidation and protects endothelial... 

PRINCEN H M, VAN DUYVENVOORDE W, BUYTENHEK R, BLONK C, TIJBURG L B, LANGIUS J A, MEINDERS A E, PUL H (1998) No effect of consiunption of green and black tea on plasma lipid antioxidant level and on LDL oxidation in smokers, Arteriosclerosis, Thrombosis and Vascular Biology, 18, 833-41. 

YING W, VINSON J A, ETHERTON T D, PROCH I, LAZARUS S A and KRIS-ETHERTON P M (2002) Effect of cocoa powder and dark chocolate on LDL oxidative susceptihihty and prostataglandin concentration in hiunans, Am J Clin Nutr, 74, 596-602. 

Recent findings from the ATBC stndy even showed that P-carotene snpple-mentation increased the post-trial risk of a hrst-ever non-fatal MI. Two secondary prevention trials, the Heart Protection Stndy and the ATBC presented similar resnlts. The former showed no association between P-carotene and fatal or non-fatal vascular events and the latter reported signihcantly increased risks of fatal coronary events in the P-carotene-snpplemented gronp. Resnlts of clinical trials focused on the effects of carotenoids on CVD biomarkers are controversial. Although carotenoid supplementation increased sernm levels,only lycopene was shown to be inversely associated with lipid, protein, DNA and LDL oxidation, and plasma cholesterol levels. - - ... 

Convincing evidence indicates that ROS generated both endogenously and also in response to diet and lifestyle factors may play a significant role in the etiology of atherosclerosis and CHD. Indeed, free radicals are responsible for LDL oxidation, which is involved in the initiation and promotion of atherosclerosis. Thus, protection from LDL oxidation by antioxidants such as carotenoids may lead to protection against human CHD. 

Experimental evidence in humans is based upon intervention studies with diets enriched in carotenoids or carotenoid-contaiifing foods. Oxidative stress biomarkers are measured in plasma or urine. The inhibition of low density lipoprotein (LDL) oxidation has been posmlated as one mechanism by which antioxidants may prevent the development of atherosclerosis. Since carotenoids are transported mainly via LDL in blood, testing the susceptibility of carotenoid-loaded LDL to oxidation is a common method of evaluating the antioxidant activities of carotenoids in vivo. This type of smdy is more precisely of the ex vivo type because LDLs are extracted from plasma in order to be tested in vitro for oxidative sensitivity after the subjects are given a special diet. 

Limited studies have focused on dietary intake of astaxanthin by humans. In a study reported by Miki, an astaxanthin-containing drink was used to protect low-density lipoprotein from oxidation (astaxanthin was administered at doses of 3.6 to 14.4 mg/day over a 2-week period). Progressive slowing of LDL oxidation with increasing doses of astaxanthin was observed and no ill effects were reported. 

Mouse peritoneal macrophages that have been activated to produce nitric oxide by 7-interferon and lipopolysac-charide were shown to oxidize LDL less readily than unactivated macrophages. Inhibition of nitric oxide synthesis in the same model was shown to enhance LDL oxidation (Jessup etal., 1992 Yates a al., 1992). It has recently been demonstrated that nitric oxide is able to inhibit lipid peroxidation directly within LDL (Ho etal., 1993c). Nitric oxide probably reacts with the propagating peroxyl radicals thus terminating the chain of lipid peroxidation. The rate constant for the reaction between nitric oxide and peroxyl radicals has recently been determined to be 1-3 X10 M" s (Padmaja and Huie, 1993). This... 

An example of an experiment in which LDL has been treated with 15-lipoxygenase and the oxidation monitored by the formation of conjugated diene is shown in Fig. 2.2. In the absence of transition metal, a rapid increase in absorbance occurs, with no lag phase, which ceases after a period of about 90 min under these conditions. If copper is added to promote LDL oxidation at this point, LDL treated with lipoxygenase oxidizes at a faster rate with a short lag phase when compared to the control. During this procedure there is only a minimal loss of a-tocopherol and so we may ascribe the shortened lag phase to the increase in lipid peroxides brought about by lipoxygenase treatment. A similar result was found when LDL was supplemented with preformed fatty acid hydroperoxides (O Leary eta/., 1992). 

Cellular lipoxygenases have been implicated as possible enzymatic mediators of endothelial cell-dependent oxidation of LDL. Inhibitors of lipoxygenase, but not cyclooxygenase, have been shown to be effective inhibitors of LDL oxidation using rabbit endothelial cells (Parthasarathy etal., 1989). Interestingly, a phospholipase A2 activity intrinsic to apo-B has also been implicated in the endothelial cell-dependent modification of LDL (Parthasarathay et al., 1985). 

Esterbauer et cil. (1992) have studied the in vitro effects of copper on LDL oxidation and have shown that there are three distinct stages in this process. In the first part of the reaction, the rate of oxidation is low and this period is often referred to as the lag phase the lag phase is apparently dependent on the endogenous antioxidant content of the LDL, the lipid hydroperoxide content of the LDL particle and the fatty acid composition. In the second or propagation phase of the reaction, the rate of oxidation is much faster and independent of the initial antioxidant status of the LDL molecule. Ultimately, the termination reactions predominate and suppress the peroxidation process. The extensive studies of Esterbauer et al. have demonstrated the relative importance of the endogenous antioxidants within the LDL molecule in protecting it from oxidative modification. 

The lag-phase measurement at 234 nm of the development of conjugated dienes on copper-stimulated LDL oxidation is used to define the oxidation resistance of different LDL samples (Esterbauer et al., 1992). During the lag phase, the antioxidants in LDL (vitamin E, carotenoids, ubiquinol-10) are consumed in a distinct sequence with a-tocopherol as the first followed by 7-tocopherol, thereafter the carotenoids cryptoxanthin, lycopene and finally /3-carotene. a-Tocopherol is the most prominent antioxidant of LDL (6.4 1.8 mol/mol LDL), whereas the concentration of the others 7-tocopherol, /3-carotene, lycopene, cryptoxanthin, zea-xanthin, lutein and phytofluene is only 1/10 to 1/300 of a-tocopherol. Since the tocopherols reside in the outer layer of the LDL molecule, protecting the monolayer of phospholipids and the carotenoids are in the inner core protecting the cholesterylesters, and the progression of oxidation is likely to occur from the aqueous interface inwards, it seems reasonable to assign to a-tocopherol the rank of the front-line antioxidant. In vivo, the LDL will also interact with the plasma water-soluble antioxidants in the circulation, not in the artery wall, as mentioned above. 

There is now strong evidence that LDL oxidation does indeed occur in vivo (see later) and strong clinical validation of the oxidation hypothesis has been achieved. The... 

Antioxidants that inhibit LDL oxidation in vitro prevent fetty streak formation in animal models (Carew etal., 1987 Kita etal., 1987 Bjorkhem etal., 1991) and others are associated with protection against coronary artery disease in population studies (Gey et al., 1991 Stampfer etal., 1993 Rimm etal., 1993). 

In contrast, Bowry and Stocker (1993) have recently proposed that a-tocopherol may act as a pro-oxidant within the LDL particle in vitro. Their studies have indicated that tocopherol-mediated LDL oxidation may take place when water-soluble alkyperoxyl radicals react with tocopheryl radicals in the absence of agents that regenerate the tocopheryl radical into a non-radical species (for example, ascorbate). Under these conditions. 

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