Aqueous Peroxyl Radicals Research Articles

Invitro free radical scavenging capacity of dimethylglycine sodium salt and its protective ability against oleic acid hydroperoxide-induced oxidative damage in IPEC-J2 cells.

The aim of the present study was to evaluate the invitro free radical scavenging capacity of dimethylglycine sodium (DMG‑Na) and its protective ability against oleic acid hydroperoxide (OAHPx)‑induced oxidative damage in IPEC‑J2 cells. Initially, the free radical scavenging activities of water‑soluble pigments (DMG‑Na, betalain, capsanthin and cyanidin‑3‑rutinoside) were measured and compared with those of Trolox. Subsequently, freshly collected swine blood was mixed with heparin and centrifuged to obtain erythrocytes. In order to induce the free radical chain oxidation in erythrocytes, the aqueous peroxyl radicals were generated by thermal decomposition of 2,2'‑azobis(2‑amidinopropane) dihydrochloride (AAPH) in oxygen. A 2% suspension of porcine erythrocytes in PBS buffer were pre‑incubated for 30min at 37˚C with DMG‑Na (32µM), followed by incubation with or without AAPH (75mM) for 5h with gentle shaking. Additionally, IPEC‑J2 cells were randomly assigned to four groups (n=6per group): Cells treated with phosphate buffered saline (PBS); cells treated with DMG‑Na (32µM); cells treated with oleic acid hydroperoxides (OAHPx, 20µM; TO group); cells treated with DMG‑Na (32µM) followed by OAHPx (20µM; DTO group). The cells were cultured in Dulbecco's modified Eagle's medium, Ham's F‑12mixture, 1.5mM HEPES, 5% (v/v) fetal bovine serum, 1% (v/v) insulin‑transferrin‑selenium mixture, 1% (v/v) penicillin‑streptomycin mixture and 2.5µg/ml fungizone (37˚C, 5% CO2). The results showed that DMG‑Na exerted the strongest free radical scavenging capacity at 0.32M from 0.08‑0.64M, and that it could prevent AAPH‑induced porcine erythrocyte hemolysis by increasing its antioxidant capacity (P<0.05). The results also demonstrated that antioxidant capacity and antioxidant‑associated gene expression increased in the DTO group relative to the TO group (P<0.05), indicating that DMG‑Na prevented the OAHPx‑induced oxidative damage in IPEC‑J2 cells by improving the antioxidant capacity and antioxidant‑associated gene expression.

Plasma lipid oxidation induced by peroxynitrite, hypochlorite, lipoxygenase and peroxyl radicals and its inhibition by antioxidants as assessed by diphenyl-1-pyrenylphosphine

Lipid oxidation has been implicated in the pathogenesis of many diseases. Lipids are oxidized in vivo by several different oxidants to give diverse products, in general lipid hydroperoxides as the major primary product. In the present study, the production of lipid hydroperoxides in the oxidation of mouse plasma induced by multiple oxidants was measured using diphenyl-1-pyrenylphosphine (DPPP) as a probe. DPPP itself is not fluorescent, but it reacts with lipid hydroperoxides stochiometrically to give highly fluorescent DPPP oxide and lipid hydroxides. The production of lipid hydroperoxides could be followed continuously in the oxidation of plasma induced by peroxynitrite, hypochlorite, 15-lipoxygenase, and peroxyl radicals with a microplate reader. A clear lag phase was observed in the plasma oxidation mediated by aqueous peroxyl radicals and peroxynitrite, but not in the oxidation induced by hypochlorite and lipoxygenase. The effects of several antioxidants against lipid oxidation induced by the above oxidants were assessed. The efficacy of antioxidants was dependent markedly on the type of oxidants. α-Tocopherol exerted potent antioxidant effects against peroxyl radical-mediated lipid peroxidation, but it did not inhibit lipid oxidation induced by peroxynitrite, hypochlorite, and 15-lipoxygenase efficiently, suggesting that multiple antioxidants with different selectivities are required for the inhibition of plasma lipid oxidation in vivo. This is a novel, simple and most high throughput method to follow plasma lipid oxidation induced by different oxidants and also to assess the antioxidant effects in biologically relevant settings.

남항(Vitis romaneti) 유래 포도잎 추출물과 Resveratrol의 항산화 및 혈행개선 효과

The purpose of this study is to investigate the effects of 80% ethanol grape leaf extract (VGLE) and resveratrol (VGLR) from Vitis romaneti on antioxidant of red blood cells (RBC) of rat and efficacy of blood flow improvement in a rat model of topical ferric chloride (FeCl3)-induced carotid artery damage. RBC oxidative hemolysis and plasma lipid peroxidation induced by the aqueous peroxyl radical generator 2,2-azobis(2-amidinopropane) dihydrochloride were significantly suppressed by VGLE or VGLR in a dose-dependent manner. The FeCl₃ treatment seriously damaged the carotid artery: the walls of the artery and blood flow. However, VGLE or VGLR administration has ameliorated the blood flow and suppressed thrombus in blood vessels. These results suggest that VGLE or VGLR ameliorates the oxidative stress of RBC and thrombosis against blood vessel damage.

Antioxidant Activity and Protective Effect of Leaf Extract from Diospyros lotus on Oxidative Stress of Red Blood Cells

This study was to evaluate the antioxidant properties of the leaf extracts of Diospyros lotus (DLE) on the chemical-induced free radical and rat red blood cell (RBC) oxidative damage in vitro. DLE were prepared by extracting with water. DLE showed the high antioxidant activities on the scavenging of 2,2-diphenyl-1-picrylhydrazyl and 2,2’-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid)-induced radicals. An antioxidant activities of DLE was similar to the reference antioxidant butylated hydroxytoluene (BHT) and (±)6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid (Trolox). Reducing power of 1,000 μg/mL DLE also was similar to the vitamin C. In RBC, oxidative hemolysis induced by the aqueous peroxyl radical generator (2,2"-Azobis (2-methylpropionamidine) dihydrochloride (AAPH)) were significantly suppressed by DLE in a dose-dependent manner. Furthermore, DLE prevented the depletion of cytosolic antioxidant glutathione in RBC damaged with AAPH. These results suggest that DLE may has value as natural product with its high quality antioxidant properties against oxidative stress.

‘In vitro’ evaluation of the antioxidant activity in relation with structure and kinetic properties of scleraldehyde

Schizophyllum commune is the world′s most extensively disseminated mushroom except in Antarctica. Schizophyllan is a polysaccharide fractions isolated from S. commune. The schizophyllan is oxidized to form scleraldehyde to introduce carbonyl group for additional functions. The relationship between structure, kinetic and antioxidant properties of scleraldehyde was studied in order to establish the molecular characteristics related to its maximum radical scavenging activity. In our earlier paper we have reported that scleraldehyde obtained from schizophyllan with the dialdehyde content of 28%. From the absorption studies, it explains there is π–π⁎ excitations involving the benzene groups. Moreover, the structural identification of scleraldehyde were characterized by bands at 1670 and 3600–3200 cm−1, corresponding to the axial deformation of the C=O bond and the angular deformation of the C–H bond respectively. The IR spectrum confirms the identity of aldehyde. The scavenging assays of scleraldehyde was found to be reliant on their reactivity and concentration. Scavenging assays performed indicate that 100 μM of scleraldehyde showed the highest antioxidant activity compared to the ascorbic acid. Furthermore, The reducing ability of the scleraldehyde was 54.81±0.92 μM of Fe(II)/g compared to the control (Ascorbic acid) respectively. The kinetic mechanism of scleraldehyde portrays a competitive inhibition against antioxidant enzymes on calculating for radical scavenging activity. By means of structure determination, kinetic assays and the antioxidant assays that hydroxyl group present in C2, C4 and C6 are the main reactive sites. The hypothesis converse that scleraldehyde can serve as scavenger of aqueous peroxyl radicals near the membrane surface of collagen.

Antioxidant effect of astragalin isolated from the leaves of Morus alba L. against free radical-induced oxidative hemolysis of human red blood cells

We evaluated the antioxidant properties of mulberry leaves extract (MLE) and flavonoids isolated from MLE. MLE was prepared by extraction with methanol. Flavonoids were analyzed by high-performance liquid chromatography. Oxidative hemolysis of normal human red blood cells (RBCs) was induced by the aqueous peroxyl radical [2,2'-Azobis (2-amidinopropane) dihydrochloride, AAPH]. MLE contained three flavonoids in the order quercetin (QC) > kaempferol (KF) > astragalin (AG). Oxidative hemolysis of RBCs induced by AAPH was suppressed by MLE, AG, KF, and QC in a time- and dose-dependent manner. MLE and these three flavonoids prevented the depletion of cystosolic antioxidant glutathione (GSH) in RBCs. AG had the greatest protective effect against AAPH-induced oxidative hemolysis and GSH depletion in RBCs.

Aqueous peroxyl radical exposure to THP-1 cells causes glutathione loss followed by protein oxidation and cell death without increased caspase-3 activity

Protein oxidation within cells exposed to oxidative free radicals has been reported to occur in an uninhibited manner with both hydroxyl and peroxyl radicals. In contrast, THP-1 cells exposed to peroxyl radicals (ROO•) generated by thermo decomposition of the azo compound AAPH showed a distinct lag phase of at least 6 h, during which time no protein oxidation or cell death was observed. Glutathione appears to be the source of the lag phase as cellular levels were observed to rapidly decrease during this period. Removal of glutathione with buthionine sulfoxamine eliminated the lag phase. At the end of the lag phase there was a rapid loss of cellular MTT reducing activity and the appearance of large numbers of propidium iodide/annexin-V staining necrotic cells with only 10% of the cells appearing apoptotic (annexin-V staining only). Cytochrome c was released into the cytoplasm after 12 h of incubation but no increase in caspase-3 activity was found at any time points. We propose that the rapid loss of glutathione caused by the AAPH peroxyl radicals resulted in the loss of caspase activity and the initiation of protein oxidation. The lack of caspase-3 activity appears to have caused the cells to undergo necrosis in response to protein oxidation and other cellular damage.

Lipid oxidation predominates over protein hydroperoxide formation in human monocyte-derived macrophages exposed to aqueous peroxyl radicals

In U937 and mouse myeloma cells, protein hydroperoxides are the predominant hydroperoxide formed during exposure to AAPH or gamma irradiation. In lipid-rich human monocyte-derived macrophages (HMDMs), we have found the opposite situation. Hydroperoxide measurements by the FOX assay showed the majority of hydroperoxides formed during AAPH incubation were lipid hydroperoxides. Lipid hydroperoxide formation began after a four hour lag period and was closely correlated with loss of cell viability. The macrophage pterin 7,8-dihydroneopterin has previously been shown to be a potent scavenger of peroxyl radicals, preventing oxidative damage in U937 cells, protein and lipoprotein. However, when given to HMDM cells, 7,8-dihydroneopterin failed to inhibit the AAPH-mediated cellular damage. The lack of interaction between 7,8-dihydroneopterin and AAPH peroxyl radicals suggests that they localize to separate cellular sites in HMDM cells. Our data shows that lipid peroxidation is the predominant reaction occurring in HMDMs, possibly due to the high lipid content of the cells.

Protection from oxidative damage using Bidens pilosa extracts in normal human erythrocytes

Bidens pilosa ( B. pilosa) is well known in Taiwan as a traditional Chinese medicine. The purpose of this study was to evaluate the ability of both the ethanol (EtOH) and ethylacetate/ethanol (EA/EtOH) extracts from the whole B. pilosa plant, to protect normal human erythrocytes against oxidative damage in vitro. It was determined that the oxidative hemolysis and lipid/protein peroxidation of erythrocytes induced by the aqueous peroxyl radical [2,2′-azobis(2-amidinopropane) dihydrochloride (AAPH)] were suppressed by both EtOH (50–150 μg/ml) and EA/EtOH (25–75 μg/ml) extracts of B. pilosa in concentration- and time-dependent manners. B. pilosa extracts also prevented the decline of superoxide dismutase (SOD) activity and the depletion of cytosolic glutathione (GSH) and ATP in erythrocytes. These results imply that B. pilosa may have protective antioxidant properties.

A Frequent Drinking of Green Tea Lowers the Levels of Endogenous Oxidative Stress in Small Intestines, Erythrocytes and Kidneys in Rats

Endogenous antioxidative potential was examined in rats given green tea as daily drinking water for 4 weeks. Rats ingested 196 μmol/kg body weight/day of catechins (sum total of 8 kinds of catechins), and plasma levels were around 0.24 μM. The green tea also contained 2.1 mM ascorbic acid, but the plasma level was similar to that in control rats drinking water, about 30 μM. The tea-drinking rats had significantly lower levels of lipid peroxidation in the small intestines and kidneys, 30% and 40% lower, respectively, than the controls, and their erythrocytes showed significant resistance to aqueous peroxyl radicals generated from azo initiator. Then, the effects of 8 catechins were examined on the antioxidative potency of ascorbic acid in in vitro. Catechins suppressed the oxidation of deoxyguanosine after the immediate exhaustion of ascorbic acid. Thus, frequent daily drinking of green tea improved the antioxidative potential of ascorbic acid.

Antioxidant properties of a novel phycocyanin extract from the blue-green alga Aphanizomenon flos-aquae

Aphanizomenon flos-aquae (AFA) is a fresh water unicellular blue-green alga (cyanophyta) rich in phycocyanin (PC), a photosynthetic pigment with antioxidant and anti-inflammatory properties. The purpose of this study was to evaluate the ability of a novel natural extract from AFA enriched with PC to protect normal human erythrocytes and plasma samples against oxidative damage in vitro. In red blood cells, oxidative hemolysis and lipid peroxidation induced by the aqueous peroxyl radical generator [2,2′-Azobis (2-amidinopropane) dihydrochloride, AAPH] were significantly lowered by the AFA extract in a time- and dose-dependent manner; at the same time, the depletion of cytosolic glutathione was delayed. In plasma samples, the natural extract inhibited the extent of lipid oxidation induced by the pro-oxidant agent cupric chloride (CuCl 2); a concomitant increase of plasma resistance to oxidation was observed as evaluated by conjugated diene formation. The involvement of PC in the antioxidant protection of the AFA extract against the oxidative damage was demonstrated by investigating the spectral changes of PC induced by AAPH or CuCl 2. The incubation of the extract with the oxidizing agents led to a significant decrease in the absorption of PC at 620 nm accompanied with disappearance of its blue color, thus indicating a rapid oxidation of the protein. In the light of these in vitro results, the potential clinical applications of this natural compound are under investigation.

Relevance of apple polyphenols as antioxidants in human plasma: contrasting in vitro and in vivo effects

Apples are a major source of flavonoids in the Western diet, and flavonoid-rich foods may help protect against chronic diseases by antioxidant mechanisms. In the present study we investigated: (1) the antioxidant capacity of representative apple polyphenols and their contribution to the total antioxidant capacity of apple extracts; (2) the effects of adding apple extract to human plasma in vitro on oxidation of endogenous antioxidants and lipids; and (3) the effects of apple consumption by humans on ex vivo oxidation of plasma antioxidants and lipids. We found that the apple-contained flavonols and flavanols, quercetin, rutin, (−)-epicatechin, and (+)-catechin, had a higher antioxidant capacity than the dihydrochalcones, phloridzin and phloretin, and the hydroxycinnamate, chlorogenic acid. However, together these apple polyphenols contributed less than 20% to the total antioxidant capacity of aqueous apple extracts. When human plasma was exposed to a constant flux of aqueous peroxyl radicals, endogenous ascorbate (70.0 ± 10.3 μM) was oxidized within 45 min of incubation, while endogenous urate (375 ± 40 μM) and α-tocopherol (24.7 ± 1.2 μM) were oxidized after ascorbate. Addition of 7.1 or 14.3 μg/ml total phenols of apple extract did not protect ascorbate from oxidation, but increased the half-life ( t 1/2) of urate from 136 ± 15 to 192 ± 16 and 208 ± 23 min, respectively ( p < 0.05 each), and t 1/2 of α-tocopherol from 141 ± 18 to 164 ± 8 min ( p = ns) and 188 ± 8 min ( p < 0.05). Lipid peroxidation started after ascorbate depletion, and addition of apple extract increased the lag time preceding detectable lipid peroxidation from 36.3 ± 3.7 to 50.9 ± 2.7 min ( p < 0.05) and 70.4 ± 4.2 min ( p < 0.001). However, when six healthy volunteers ate five apples and plasma was obtained up to 4 h after apple consumption, no significant increases in the resistance to oxidation of endogenous urate, α-tocopherol, and lipids were found. Thus, despite the high antioxidant capacity of individual apple polyphenols and apple extracts and the significant antioxidant effects of apple extract added to human plasma in vitro, ingestion of large amounts of apples by humans does not appear to result in equivalent in vivo antioxidant effects of apple polyphenols.

Protection of oxidative damage by aqueous extract from Antrodia camphorata mycelia in normal human erythrocytes

Antrodia camphorata ( A. camphorata) is well known in Taiwan as a traditional Chinese medicine. The purpose of this study was to evaluate the ability of aqueous extract from A. camphorata mycelia to protect normal human erythrocytes against oxidative damage in vitro. Oxidative hemolysis and lipid/protein peroxidation of erythrocytes induced by the aqueous peroxyl radical [2,2′-Azobis(2-amidinopropane) dihydrochloride, AAPH] were suppressed by A. camphorata mycelia in a time-and concentration-dependent manner. A. camphorata mycelia also prevented the depletion of cytosolic antioxidant glutathione (GSH) and ATP in erythrocytes. Moreover, cultured human endothelial cell damage induced by AAPH was suppressed by A. camphorata mycelia. Interestingly, A. camphorata mycelia exhibited significant cytotoxicity against leukemia HL-60 cells but not against cultured human endothelial cells. These results imply that A. camphorata mycelia may have protective antioxidant and anticancer properties.

A method to measure the oxidizability of both the aqueous and lipid compartments of plasma

The lipophilic radical initiator (MeO-AMVN) and the fluorescent probe C11BODIPY581/591 (BODIPY) were used to measure the lipid compartment oxidizability of human plasma. Aqueous plasma oxidizability was initiated by the aqueous peroxyl radical generator, AAPH, and 2′,7′-dichlorodihydrofluorescein (DCFH) was employed as the marker of the oxidative reaction. The distribution in aqueous and lipid compartments of the two radical initiators was determined by measuring the rate of consumption of the plasma hydrophilic and lipophilic endogenous antioxidants. In the presence of AAPH (20 mM), the order of consumption was: ascorbic acid > α-tocopherol > uric acid > β-carotene, indicating a gradient of peroxyl radicals from the aqueous to the lipid phase. When MeO-AMVN was used (2mM), β-carotene was consumed earlier than uric acid and almost at the same time as α-tocopherol, reflecting the diffusion and activation of MeO-AMVN in the lipophilic phase. The rate of BODIPY oxidation (increase in green fluorescence) significantly increased after the depletion of endogenous α-tocopherol and β-carotene, whereas it was delayed for 180 min when AAPH was used instead of MeO-AMVN. The measurement of lipid oxidation in plasma was validated by adding to plasma the two lipophilic antioxidants, α-tocopherol and β-carotene, whose inhibitory effects on BODIPY oxidation were dependent on the duration of the preincubation period and hence to their lipid diffusion. DCFH oxidation induced by AAPH only began after uric acid, the main hydrophilic plasma antioxidant, was consumed. In contrast, when MeO-AMVN was used, DCFH oxidation was delayed for 120 min, indicating its localization in the aqueous domain. In summary, the selective fluorescence method reported here is capable of distinguishing the lipophilic and hydrophilic components of the total antioxidant capacity of plasma.

Inhibitory effect of a quercetin metabolite, quercetin 3-O-beta-D-glucuronide, on lipid peroxidation in liposomal membranes.

To study the antioxidant activity of quercetin 3-O-beta-D-glucuronide (Q3GA), which is one of the quercetin metabolites in the blood after intake of quercetin-rich food, the inhibitory effect of Q3GA on lipid peroxidation was estimated using phosphatidylcholine large unilamellar vesicles (PC LUV) as a biomembrane model. Iron ion, an aqueous peroxyl radical generator, a peroxynitrite generator, or lipoxygenase was used as the inducer of lipid peroxidation. In all cases, Q3GA inhibited lipid peroxidation significantly, although its inhibitory effect was lower than that of quercetin aglycon. The ultrafiltration of PC LUV containing Q3GA revealed that Q3GA has low but significant affinity with the membranes of phospholipid bilayers. It is therefore likely that Q3GA acts as an efficient antioxidant in membranous lipid peroxidation through its localization in the phospholipid bilayer. This conjugated quercetin metabolite seems to retain the ability to protect cellular and subcellular membranes from peroxidative attack by reactive oxygen species and peroxidative enzymes.

Red wine antioxidants bind to human lipoproteins and protect them from metal ion-dependent and -independent oxidation.

Plant-derived polyphenols may exert beneficial effects on atherosclerosis and cardiovascular diseases, in part, because of their antioxidant properties. In this study we compared the effects of unbound (free) and lipoprotein-associated red wine components on in vitro antioxidant protection of human low-density lipoprotein (LDL). Preincubation of LDL (1 mg protein/mL) with 0-2.5% (v/v) red wine for 3 h at 37 degrees C followed by gel filtration to remove unbound red wine components resulted in a dose-dependent, up to 4-fold increase in LDL-associated antioxidant capacity (measured as Trolox equivalents). Similar results were obtained with high-density lipoprotein (HDL) and bovine serum albumin (BSA). Furthermore, LDL was subjected to oxidation by copper and aqueous peroxyl radicals (2,2'-azobis[2-amidinopropane] dihydrochloride, AAPH). Under both types of oxidative stress, LDL-associated and free red wine components significantly decreased oxidation of the lipoprotein's protein moiety (assessed by tryptophan fluorescence) and lipid moiety (assessed by thiobarbituric acid-reactive substances and conjugated dienes). Similar protective effects of red wine components were observed against HDL oxidation. In contrast, red wine exerted a pro-oxidant effect on copper-induced oxidation of BSA tryptophan residues, while protecting them from AAPH-induced oxidation. Ascorbate strongly enhanced the protective effect of red wine against copper-induced LDL oxidation, and had an additive effect against AAPH-induced oxidation. Our data indicate that red wine components bind to LDL and HDL and protect these lipoproteins from metal ion-dependent and -independent protein and lipid oxidation.

Glucose enhancement of LDL oxidation is strictly metal ion dependent

Recent evidence suggests that lipoprotein oxidation is increased in diabetes, however, the mechanism(s) for such observations are not clear. We examined the effect of glucose on low-density lipoprotein (LDL) oxidation using metal ion–dependent and –independent oxidation systems. Pathophysiological concentrations of glucose (25 mM) enhanced copper-induced LDL oxidation as determined by conjugated diene formation or relative electrophoretic mobility (REM) on agarose gels. Similarly, iron-induced LDL oxidation was stimulated by glucose resulting in 4- to 6-fold greater REM than control incubations without glucose. In contrast, glucose had no effect on metal ion–independent LDL oxidation by aqueous peroxyl radicals. The effect of glucose on metal ion–dependent LDL oxidation was associated with enhanced reduction of metal ions, and in the case of iron-induced LDL oxidation, was completely inhibited by superoxide dismutase. The effect of glucose was mimicked by other reducing sugars, such as fructose and mannose, and the extent to which each sugar enhanced LDL oxidation was closely linked to its metal ion–reducing activity. Thus, promotion of LDL oxidation by glucose is specific for metal ion–dependent oxidation and involves increased metal ion reduction. These results provide one potential mechanism for enhanced LDL oxidation in diabetes.

Coenzyme Q improves LDL resistance to ex vivo oxidation but does not enhance endothelial function in hypercholesterolemic young adults

Oxidative modification of low-density lipoprotein (LDL) may cause arterial endothelial dysfunction in hyperlipidemic subjects. Antioxidants can protect LDL from oxidation and therefore improve endothelial function. Dietary supplementation with coenzyme Q (CoQ 10) raises its level within LDL, which may subsequently become more resistant to oxidation. Therefore, the aim of this study was to assess whether oral supplementation of CoQ 10 (50 mg three times daily) is effective in reducing ex vivo LDL oxidizability and in improving vascular endothelial function. Twelve nonsmoking healthy adults with hypercholesterolemia (age 34 ± 10 years, nine women and three men, total cholesterol 7.4 ± 1.1 mmol/l) and endothelial dysfunction (below population mean) at baseline were randomized to receive CoQ 10 or matching placebo in a double-blind crossover study (active/placebo phase 4 weeks, washout 4 weeks). Flow-mediated (FMD, endothelium-dependent) and nitrate-mediated (NMD, smooth muscle-dependent) arterial dilatation were measured by high-resolution ultrasound. CoQ 10 treatment increased plasma CoQ 10 levels from 1.1 ± 0.5 to 5.0 ± 2.8 μmol/l ( p = .009) but had no significant effect on FMD (4.3 ± 2.4 to 5.1 ± 3.6 %, p = .99), NMD (21.6 ± 6.1 to 20.7 ± 7.8 %, p = .38) or serum LDL-cholesterol levels ( p = .51). Four subjects were selected randomly for detailed analysis of LDL oxidizability using aqueous peroxyl radicals as the oxidant. In this subgroup, CoQ 10 supplementation significantly increased the time for CoQ 10H 2 depletion upon oxidant exposure of LDL by 41 ± 19 min ( p = .04) and decreased the extent of lipid hydroperoxide accumulation after 2 hours by 50 ± 37 μmol/l ( p = .04). We conclude that dietary supplementation with CoQ 10 decreases ex-vivo LDL oxidizability but has no significant effect on arterial endothelial function in patients with moderate hypercholesterolemia.

Antioxidant defenses in rat intestine and mesenteric lymph

Dietary oxysterols can reach the circulation and this may contribute to atherosclerosis, where lipid oxidation is thought to be important. There is also evidence that, in rats,peroxidized lipids are absorbed and transported into lymph [Aw TY, Williams MW, Gray L. Absorption and lymphatic transport of peroxidized lipids by rat small intestine in vivo: role of mucosal GSH. Am J Physiol 1992; 262: G99–G106], although the method used to detect lipid peroxides lacked specificity. We tested whether intragastric administration of vegetable oils containing triglyceride hydroperoxides (TG-OOH) to rats resulted in detectable lipid hydroperoxides in mesenteric lymph. Using sensitive HPLC with postcolumn chemiluminescence detection, we were unable to detect hydroperoxides of triglycerides, cholesterylesters or phospholipids during the course of lipid absorption, and lymph levels of ascorbate, urate, α-tocopherol and ubiquinol-9 did not change significantly. By contrast, we observed a striking reducing activity judged by the efficient reduction of administered ubiquinones-9 and -10 to the corresponding ubiquinols. Exposure of rat lymph and isolated chylomicrons to aqueous peroxyl radicals revealed patterns of antioxidant consumption and lipid hydroperoxide formation similar to those described previously for human extravascular fluids and isolated lipoproteins, respectively. In particular, rates of TG-OOH formation in lymph and chylomicrons were very low to undetectable as long as ascorbate and/or ubiquinols were present, but subsequently proceeded in a chain reaction despite the presence of α-tocopherol. These studies demonstrate that rat intestine and mesenteric lymph possess efficient antioxidant defenses against preformed lipid hydroperoxides and (peroxyl) radical mediated lipid oxidation. We conclude that dietary lipid hydroperoxides or postprandial oxidation of lipids are not likely to contribute to these particular forms of oxidized lipids in circulation and aortic tissue.

Prooxidant and antioxidant properties of Trolox C, analogue of vitamin E, in oxidation of low-density lipoprotein

Trolox C (Trolox), a water-soluble analogue of vitamin E lacking the phytyl chain, was investigated with respect to its effect on the oxidation of low-density lipoprotein (LDL). Trolox was added at different time points of LDL oxidation induced by Cu2+ and aqueous peroxyl radicals. In the case of Cu2+ -induced LDL oxidation, the effect of Trolox changed from antioxidant to prooxidant when added at later time points during oxidation; this transition occurred whenever α-tocopherol was just consumed in oxidizing LDL. Thus, in the case of Cu2+-dependent LDL oxidation, the presence of lipophilic antioxidants in the LDL particle is likely to be a prerequisite for the antioxidant activity of Trolox.When oxidation was induced by peroxyl radicals, as a model of metal-independent oxidation, the effect of Trolox was always antioxidant, suggesting the importance of Cu2+/Cu+ redox-cycling in the prooxidant mechanism of Trolox. Our data suggest that, in the absence of significant amounts of lipophilic antioxidants, LDL becomes highly susceptible to oxidation induced by transition metals in the presence of aqueous reductants.