维生素C在低密度脂蛋白氧化修饰中作用的体外实验

目的：研究维生素C（VC）在预防低密度脂蛋白（LDL）氧化修饰中的作用。方法：在Cu^2 介导的无细胞体系中分别加入VC10,50,100,200μmol/L,巨噬细胞体中分别加入VC50,100,200μmol/L,以维生素（200μmol/L）为阳性对照,不添加维生素C组（VC0）为阴性对照,测定荧光物质（lipofusin),硫代巴比妥酸反应物质（TBARS）,LDL电泳迁移率（Rf),氧化型低密度脂蛋白（Ox-LDL),LDL氧化过程中的停滞时间等LDL的氧化修饰情况。结果：Cu^2 介导的无细胞体系中,高剂量VC（100,200μmol/L）在3,6,9h均能发挥其抗氧化作用,抑制TBARS,Ox-LDL生成,而VC低剂量（10,50μmol/L）则无此作用,相反在3h表现出促Ox-LDL生成作用,在巨噬细胞体系中,高剂量组（100,200μmol/L)能显著降低荧光物质,TBARS和LDL电泳迁移率（Rf),延长LDL氧化过程中的停滞时间,并存在剂量反应关系。结论：VC在LDL氧化修饰中具有双重作用,低剂量时能促进LDL氧化,高剂量时表现出抗氧化作用。     

Do iron and vitamin C co-supplementation influence platelet function or LDL oxidizability in healthy volunteers?

OBJECTIVE: To examine the effect of co-supplementation with iron and vitamin C on antioxidant status, platelet function and low density lipoprotein oxidation in normal healthy volunteers. DESIGN: The study was carried out with two groups of 20 subjects each acting as their own control, comparing presupplemention with postsupplemention. One group was supplemented with iron and the RDA level of vitamin C and the second group with iron and 260 mg/d vitamin C. SETTING: The International Antioxidant Research Centre, The Guy's, King's College and St Thomas's School of Biomedical Science, Guy's Campus, London. SUBJECTS: Forty normal healthy volunteers, recruited from the staff of the Medical School and Hospital in which two volunteers withdrew during the study. INTERVENTIONS: Subjects in both studies were randomly assigned to one of two groups (5 males and 5 females group) and received supplements containing iron (14 mg/d) and either 60 mg/d (Group A) or 260 mg/d (Group B) vitamin C for 12 wk. Blood samples were taken at 6 wk and 12 wk, and prior to supplementation and analysed for iron and antioxidant status (transferrin bound iron, vitamin C and E, and beta-carotene levels) in both studies. Samples from the first study were analysed for the susceptibility of LDL isolated from plasma to Cu2+-induced oxidation and samples from the second for platelet function. RESULTS: Transferrin-bound iron was significantly increased (P < 0.05) at 12 wk, in Group A subjects (from 14.9+/-5.3 micromol/1 to 19.5+/-2.3 micromol/l; mean+/-s.d.; n=19), whereas those in Group B showed a significant increase (P < 0.05) after 6 wk (from 15.8+/-4.5 micromol/l to 20.4+/-6.6 micromol/l; n = 19) which decreased at 12 wk (16.3+/-5.0 micromol/l). Plasma total ascorbate significantly increased from an initial level of 59.3+/-21.3 micromol/l to 87.6+/-29.0 micromol/l after 6 wk and 81.7+/-11.4 micromol/l after 12 wk following the Group B supplementation, but only after 12 wk in Group A (from 64.0+/-24.8 micromol/l to 77.2+/-13.2 micromol/l). Plasma alpha-tocopherol concentrations were significantly increased after 6 wk and 12 wk with both levels of supplementation (from 24.2+/-5.71 micromol/l Group A and 23.4+/-5.3 micromol/l Group B to 26.3+/-5.5 micromol/l and 25.71+/-4.7 micromol/1 respectively at 12wk). The mean lag phase to oxidation of low density lipoprotein (LDL) was significantly increased in subjects in Group B after 12 wk ingestion of iron and 260 mg vitamin C (from 80.0+/-14.8 min to 97.2+/-16.9 min; n = 9). Platelet sensitivity to ADP-induced aggregation was significantly decreased (P < 0.05) by 12 wk in Group A (from EC50 2.3 < or = 1.3 microM to 3.7+/-2.2 microM; n = 10), whereas those receiving higher vitamin C showed a significant decrease (P < 0.05; from EC50 1.9+/-0.6 microM to 3.1+/-1.8 microM) after 6wk which subsequently increased towards presupplemental levels (2.6+/-1.6 microM). Platelets from the latter subjects showed a significant reduction in ADP-induced ATP secretion at both 6wk and 12 wk. CONCLUSION: The results show modest beneficial effects on LDL oxidation and platelet function following supplementation with iron and vitamin C. No evidence for pro-oxidant effects was observed.     

Flavonoids from almond skins are bioavailable and act synergistically with vitamins C and E to enhance hamster and human LDL resistance to oxidation

Consumption of tree nuts such as almonds has been associated with a reduced risk of coronary heart disease. Flavonoids, found predominantly in the skin of almonds, may contribute to their putative health benefit, but their bioactivity and bioavailability have not previously been studied. Almond skin flavonoids (ASF) were extracted with HCl:H2O:methanol (1:19:80) and their content of catechins and flavonols identified by HPLC with electrochemical detection. ASF bioactivity was assessed in vitro by their capacity to increase the resistance of human LDL to oxidation induced by 10 micromol/L Cu2+. ASF from 0.18 to 1.44 mumol gallic acid equivalent (GAE)/L increased the lag time to LDL oxidation in a dose-dependent manner (P < or = 0.0001). Combining ASF with vitamin E or ascorbic acid extended the lag time >200% of the expected additive value (P < or = 0.05). The bioavailability and in vivo antioxidant activity of 40 micromol ASF were examined in BioF1B hamsters. Peak plasma concentrations of catechin, epicatechin, and flavonols (quercetin, kaempferol, and isorhamnetin) occurred at 60, 120, and 180 min, respectively. The concentration of isorhamnetin was significantly elevated in liver at 180 min. Absorbed ASF enhanced the ex vivo resistance of hamster LDL collected at 60 min to oxidation by 18.0% (P = 0.028), and the in vitro addition of 5.5 micromol/L vitamin E synergistically extended the lag time of the 60-min sample by 52.5% (P < or = 0.05). Thus, ASF possess antioxidant capacity in vitro; they are bioavailable and act in synergy with vitamins C and E to protect LDL against oxidation in hamsters.     

The effect of vitamin E and vitamin C supplementation on LDL oxidizability and neutrophil respiratory burst in young smokers

OBJECTIVE: The purpose of this study was to determine the effect of vitamin E and/or vitamin C supplementation on low-density lipoprotein (LDL) oxidizability and neutrophil (PMN) superoxide anion production in young smokers. METHODS: Thirty smokers with a <5 pack-year history were randomly assigned to take placebo; vitamin C (1 g/day); vitamin E (400 IU/day), or both vitamins in a double-blind fashion. Subjects took the supplements for 8 weeks. At weeks 0 and 8, blood was collected for isolation of LDL and PMN, and for antioxidant vitamin analysis. LDL was oxidized with a copper (Cu) catalyst, and oxidation was measured by formation of conjugated dienes over a 5-hour time course. Lag times and maximum oxidation rates were calculated from the time course data. PMN superoxide anion release was assessed by respiratory burst after stimulation with phorbol ester and opsonized zymosan, and their ability to oxidize autologous LDL following treatment with the above stimuli was measured with the conjugated diene assay. RESULTS: Subjects who received vitamin E alone had a significant increase in the lag phase of Cu-catalyzed LDL oxidation (week 0, 118+/-31 min vs. week 8, 193+/-80 min, mean +/- SD, p < 0.05), whereas the vitamin C and placebo groups had no changes in LDL oxidation kinetics. The group receiving both vitamins E and C had a significant reduction in oxidation rate (week 0. 7.4+/-2.3 vs. week 8, 5.1+/-2.1, p < 0.05). There were no significant changes for any group in PMN superoxide anion production or PMN LDL oxidation after stimulation with either phorbol ester or opsonized zymosan. Plasma and LDL vitamin E concentrations were significantly increased in both groups that received vitamin E. The subjects who received vitamin C alone had no significant change in plasma vitamin C concentrations; however, when data were pooled from both groups who received vitamin C, the increases were significant. CONCLUSION: Vitamin E supplementation of young smokers was effective in reducing Cu-catalyzed LDL oxidizability; however, vitamin E and/or C supplementation showed few significant effects on the more physiologically relevant PMN function. This casts doubt on the ability of antioxidant supplementation to reduce oxidative stress in smokers in vivo. Therefore, smoking cessation remains the only means by which young smokers can prevent premature coronary heart disease.     

Pomegranate wine has greater protection capacity than red wine on low-density lipoprotein oxidation

Although there is a large body of evidence on the main role of red wine in protection of low-density lipoprotein (LDL) against oxidation, there are few data on the role of pomegranate juice, which has high phenolic content. We conducted this study considering the possible importance of pomegranate wine as an antioxidant and in order to make a comparison between red and pomegranate wines. The phenol levels of pomegranate and red wines (4,850 mg/L gallic acid equivalents and 815 mg/L gallic acid equivalents, respectively) were in accordance with their total antioxidant activity (39.5% and 33.7%, respectively). Both wines decreased LDL-diene levels following a 30-minute incubation period compared with controls (145 +/- 3.2 micromol/mg of LDL protein). However, pure pomegranate wine demonstrated a greater antioxidant effect (P < .01) on diene level (110 +/- 4.6 micromol/mg of LDL protein) than pure red wine (124 +/- 3.2 micromol/mg of LDL protein). In conclusion, we suggest that pomegranate wine has potential protective effects toward LDL oxidation, and it may be a dietary choice for people who prefer fruit wines.     

Avenanthramides and phenolic acids from oats are bioavailable and act synergistically with vitamin C to enhance hamster and human LDL resistance to oxidation

The intake of phenolic acids and related polyphenolic compounds has been inversely associated with the risk of heart disease, but limited information is available about their bioavailability or mechanisms of action. Polyphenolics, principally avenanthramides, and simple phenolic acids in oat bran phenol-rich powder were dissolved in HCl:H(2)O:methanol (1:19:80) and characterized by HPLC with electrochemical detection. The bioavailability of these oat phenolics was examined in BioF1B hamsters. Hamsters were gavaged with saline containing 0.25 g oat bran phenol-rich powder (40 micromol phenolics), and blood was collected between 20 and 120 min. Peak plasma concentrations of avenanthramides A and B, p-coumaric, p-hydroxybenzoic, vanillic, ferulic, sinapic, and syringic acids appeared at 40 min. Although absorbed oat phenolics did not enhance ex vivo resistance of LDL to Cu(2+)-induced oxidation, in vitro addition of ascorbic acid synergistically extended the lag time of the 60-min sample from 137 to 216 min (P < or = 0.05), unmasking the bioactivity of the oat phenolics from the oral dose. The antioxidant capability of oat phenolics to protect human LDL against oxidation induced by 10 micromol/L Cu(2+) was also determined in vitro. Oat phenolics from 0.52 to 1.95 micromol/L increased the lag time to LDL oxidation in a dose-dependent manner (P < or = 0.0001). Combining the oat phenolics with 5 micromol/L ascorbic acid extended the lag time in a synergistic fashion (P < or = 0.005). Thus, oat phenolics, including avenanthramides, are bioavailable in hamsters and interact synergistically with vitamin C to protect LDL during oxidation.     

Effects of supplementation with folic acid and antioxidant vitamins on homocysteine levels and LDL oxidation in coronary patients

Hyperhomocysteinemia is an important cardiovascular risk factor. Serum homocysteine levels are specially dependent on folate nutritional status. In addition, the oxidative modification of low-density lipoproteins (LDLs) in the endothelial microenvironment is a damaging factor that can be modified with fat-soluble antioxidant vitamins. The present study was done to assess the effect of a supplementation of folic acid and antioxidant vitamins on homocysteine levels and in vitro LDL oxidation in patients with coronary artery disease. Twenty-three patients with angiographically proven coronary artery disease were given supplements for 15 d consisting of one capsule twice a day of a multivitamin preparation containing 0.65 mg folic acid, 150 mg alpha-tocopherol, 150 mg ascorbic acid, 12.5 mg beta-carotene, and 0.4 microgram vitamin B12. Serum lipids, vitamin and homocysteine levels, and in vitro LDL oxidation were measured before and after the supplementation period. During the supplementation period, serum folate levels increased from 5.0 +/- 1.5 to 10.8 +/- 3.8 ng/mL (P < 0.001), vitamin B12 increased from 317.4 +/- 130.4 to 334.5 +/- 123.8 pg/mL (P < 0.05), and alpha-tocopherol increased from 8.2 +/- 5.1 to 13.7 +/- 7.9 mg/L (P < 0.001). Serum homocysteine levels decreased from 8.7 +/- 4.3 to 6.3 +/- 2.2 mumol/L (P < 0.001). In vitro LDL oxidation decreased from 2.6 +/- 1.1 to 1.6 +/- 1.1 nmol malondialdehyde/mg protein (P < 0.001). In comparing patients with healthy controls, basal levels of folate were lower in the patients, whereas vitamin B12, alpha-tocopherol, and homocysteine levels were similar. No changes in serum lipid levels or body weight were observed. In conclusion, a short-term supplementation with folic acid and antioxidant vitamins can reduce serum homocysteine levels and in vitro LDL oxidation in patients with coronary artery disease.     

A comparison between the antioxidant and peroxynitrite-scavenging functions of the vitamin E metabolites alpha- and gamma-carboxyethyl-6-hydroxychromans

Carboxyethyl-6-hydroxychromans (CEHC) are vitamin E metabolites with proposed in vitro antioxidant function. In this study we compared the antioxidant potency of the two main CEHC metabolites found in biological fluids (i.e.. alpha-CEHC and gamma-CEHC) using two different experimental models of lipid oxidation: 1) plasma diluted 1/50 vol/vol in phosphate buffered saline (PBS) exposed to 50 micro microM Cu2+ ions, and 2) LDL (100 microg of proteins) exposed to different pro-oxidants as 2.5 microM Cu2+, 1 mM of the water soluble peroxyl radical generator 2,2'-Azobis(2-amidinopropane) hydrochloride (AAPH) and human macrophages (4 x 10(5) cells). Moreover, the two CEHC homologues were assessed for the inhibitory effect on the peroxynitrite (ONOO-)-induced nitration of tyrosine (Tyr). The results showed that in the concentration range 0.015-5 microM the CEHC metabolites and the hydrosoluble analogue Trolox exert similar concentration-dependent inhibition of the Cu2+-induced lipid oxidation of plasma. After in vitro exposure to tert-butyl hydroperoxide/Fe2+, CEHC formed chromanoxyl radicals with electron spin resonance spectra matching exactly those of their parent tocopherols. The LDL oxidation induced by AAPH or Cu2+ was significantly and similarly inhibited by 1 microM of both the CEHC homologues and Trolox. gamma-CEHC showed a slight but significantly higher inhibition of the macrophage-induced low-density lipoprotein (LDL) oxidation than alpha-CEHC. Both the CEHC homologues inhibit Tyr nitration induced by ONOO. However, gamma-CEHC produced a slightly greater inhibitory effect than (alpha-CEHC through the formation of the nitrated congener 5-nitro-gamma-CEHC. In all the systems under investigation, low nanomolar concentrations of CEHC (i.e., the concentration range in the blood of subjects with normal dietary intake of vitamin E) produced feeble antioxidant effects. In conclusion, gamma-CEHC and alpha-CEHC show similar concentration-dependent inhibition of plasma and LDL lipid oxidation. gamma-CEHC has a fairly higher potency than alpha-CEHC as ONOO- scavenger through the formation of 5-nitro-gamma-CEHC. CEHC metabolites show the same in vitro antioxidant chemistry of their parent tocopherols, but the characteristic hydrophilicity of these metabolites could result in different biopotency and roles. Further studies are needed to clarify whether CEHC could contribute to the antioxidant network in biological fluids and tissues.     

Alpha-tocopherol distribution in lipoproteins and anti-inflammatory effects differ between CHD-patients and healthy subjects

OBJECTIVE: The purpose of this study was to investigate the dose-dependent effects of RRR-alpha-tocopherol supplementation in coronary heart disease (CHD) patients and healthy subjects on plasma alpha-tocopherol levels, plasma lipoprotein distribution, LDL oxidation, and inflammatory plasma markers. METHODS: 12 patients with coronary heart disease and 12 healthy subjects were supplemented with increasing dosages of RRR-alpha-tocopherol at 100, 200 and 400 mg/day for a period of 3 weeks per dose. Lipoproteins were separated by FPLC and ultracentrifugation. Alpha-tocopherol was measured by HPLC. Resistance of LDL to oxidation was determined by reading the absorption at 234 nm after CuCl2-induced oxidation. Clinical chemistry and inflammatory markers were measured on automated analysis systems. RESULTS: Plasma alpha-tocopherol concentrations at baseline were comparable between CHD-patients and healthy subjects (21.7 +/- 4.7 micromol/L and 25.8 +/- 7.6 micromol/L, respectively). CHD-patients showed a significant increase (59%) of plasma alpha-tocopherol concentrations to 34.6 +/- 9.8 micromol/L at a dosage of 100 mg/day RRR-alpha-tocopherol, whereas healthy subjects showed a significant (54%) increase to 39.7 +/- 6.1 micromol/L only with 400 mg/day RRR-alpha-tocopherol. In addition, CHD-patients showed a significantly increased enrichment of alpha-tocopherol in VLDL. Supplementation (200 mg/day) caused a significant decrease of the acute phase plasma proteins C-reactive protein (CRP) (-65%) and fibrinogen (-24%). CONCLUSION: Our data demonstrate that CHD-patients require lower dosages of alpha-tocopherol supplementation than healthy subjects to exert biological effects on plasma lipoproteins and acute phase response.     

Orange juice ingestion and supplemental vitamin C are equally effective at reducing plasma lipid peroxidation in healthy adult women

OBJECTIVE: To directly examine the contribution of vitamin C to the antioxidant potential of fruits and vegetables, the antioxidant effect of orange juice consumption (8 and 16 fl. oz.) was compared to the antioxidant effect of supplemental vitamin C (dosage equivalent to that supplied by 8 fl. oz. of orange juice). METHODS: Subjects (n = 11; 28.6 +/- 2.1 years) received each treatment in a 3 x 3 randomized crossover design, and each two-week treatment was preceded by a two-week washout. During the entire trial, subjects restricted fruit and vegetable consumption to < or =3 servings per day except the vitamin C-rich foods (items containing >20 mg/serving), which were restricted to < or =3 servings per week. A fasting blood sample was collected at the end of each washout and each treatment period. RESULTS: Following washouts, plasma vitamin C and lipid peroxidation (plasma TBARS) were similar by treatment group and averaged 25.4 +/- 3.6 micromol/L and 3.82 +/- 0.10 nmol/mL respectively. Plasma vitamin C concentrations were similar following each treatment period, 37.9 +/- 8.1, 45.8 +/- 9.4, and 38.3 +/- 12.4 micromol/L for the 8 and 16 fl. oz. orange juice treatments and the supplement treatment, respectively. All intervention treatments reduced plasma TBARS as compared to pretreatment values: -47% (p = 0.013), -40% (p = 0.083), and -46% (p = 0.015) for the 8 and 16 fl. oz. orange juice treatments and supplement treatment respectively. CONCLUSIONS: These data indicate that the regular consumption of 8 fl. oz. orange juice or supplemental vitamin C ( approximately 70 mg/day) effectively reduced a marker of lipid peroxidation in plasma.     

Vitamin C attenuates hypochlorite-mediated loss of paraoxonase-1 activity from human plasma

Paraoxonase 1 (PON1) is a cardioprotective enzyme associated with high-density lipoprotein (HDL). We tested the hypothesis that vitamin C protects HDL and PON1 from deleterious effects of hypochlorous acid, a proinflammatory oxidant. In our experiments, HDL (from human plasma) or diluted human plasma was incubated with hypochlorite in either the absence (control) or presence of vitamin C before measuring chemical modification and PON1 activities. Vitamin C minimized chemical modification of HDL, as assessed by lysine modification and accumulation of chloramines. In the absence of vitamin C, chloramines accumulated to 114 ± 4 μmol/L in HDL incubated with a 200-fold molar excess of hypochlorite; but addition of vitamin C (200 μmol/L) limited formation to 36 ± 6 μmol/L (P < .001). In plasma exposed to hypochlorite, IC₅₀ values of 1.2 ± 0.1, 9.5 ± 1.0, and 5.0 ± 0.6 mmol/L were determined for PON1's phosphotriesterase, arylesterase, and (physiologic) lactonase activities, respectively. Vitamin C lessened this inhibitory effect of hypochlorite on PON1 activities. In plasma supplemented with vitamin C (400 μmol/L), PON1 phosphotriesterase activity was 72% ± 17% of normal after incubation with hypochlorite (2 mmol/L), compared with 42% ± 6% for unsupplemented plasma (P < .05). Similar effects were seen for other PON1 activities. In some experiments, vitamin C also appeared to reverse hypochlorite-mediated loss of PON1 phosphotriesterase activity; but this effect was not observed for the other PON1 activities. In conclusion, vitamin C attenuated hypochlorite-mediated loss of PON1 activity in vitro and may, therefore, preserve cardioprotective properties of HDL during inflammation.     

银杏叶对低密度脂蛋白氧化修饰抑制作用的时效研究

目的　探讨银杏叶不同有效成分 [总提取物EGB76 1 ;银杏叶总黄酮 (GF) ;银杏叶总内酯 (GT) ]对Fe2 + 或Cu2 + 诱导的人血浆低密度脂蛋白 (LDL)氧化修饰的抑制作用与时间的变化关系。方法　使用荧光分光光度法测定了LDL在Fe2 + 或Cu2 + 诱导下 ,分别加入上述三种有效成分 ,作用 1、2、4、6h后自身荧光物质(LF)、丙二醛 (MDA)和维生素E(VitE)含量。结果　三种有效成分对LF和MDA的生成及VitE的消耗有抑制作用 ,抑制作用在 4h内随作用时间延长逐渐明显 ,6h时抑制作用明显减弱。结论　银杏叶对低密度脂蛋白氧化修饰抑制作用具有一定的时效关系     

Effect of in-vivo supplementation with low-dose vitamin E on susceptibility of low-density lipoprotein and high-density lipoprotein to oxidative modification.

We have examined the effects of in-vivo supplementation with low-dose vitamin E on the susceptibility of low-density lipoprotein (LDL) and high-density lipoprotein (HDL) to oxidative modification, and compared the oxidizability of HDL with that of LDL.

Normal humans (n = 8) ingested vitamin E (150 mg/day for 1 week, followed by 300 mg/day for 3 weeks) in divided doses with meals. The subjects did not use any medications or vitamins before being enrolled in this study. Fasting blood samples were drawn before and at the end of supplementation. LDL and HDL were separated by sequential ultracentrifugation and susceptability to copper-mediated oxidation was measured.

After vitamin E supplementation, vitamin E content of LDL increased 1.9-fold and that of HDL increased 1.8-fold. Lag time before initiation of LDL oxidation lengthened significantly (+20%, p < 0.01), and the propagation rate of LDL decreased significantly (?10%, p < 0.05). The lag time of HDL oxidation did not change significantly, but the propagation rate of HDL oxidation decreased significantly (?24%, p < 0.001). The lag time of HDL oxidation was shorter than that of LDL. HDL contained the same or higher concentrations of vitamin E relative to lipid mass as LDL, but contained lower concentration of CoQ10 relative to lipid mass and fewer molecules of vitamin E and beta-carotene per particle than LDL.

We conclude that in-vivo supplementation of low-dose vitamin E protects LDL against oxidative modification and decreases the propagation rate of HDL oxidation significantly. We suggest that supplementation with low-dose vitamin E would be beneficial for ameliorating atherosclerosis.     

Flaxseed oil supplementation does not affect plasma lipoprotein concentration or particle size in human subjects

alpha-Linolenic acid (ALA) is a major dietary (n-3) fatty acid. Some clinical trials with ALA supplementation have shown reduced cardiovascular risk; however the specific cardioprotective mechanism is not known. We studied the effects of daily supplementation with ALA derived from flaxseed oil on concentrations of plasma LDL cholesterol, HDL cholesterol, intermediate density lipoprotein cholesterol, and lipid particle sizes. In a randomized double-blind trial, 56 participants were given 3 g/d of ALA from flaxseed oil in capsules (n = 31) or olive oil containing placebo capsules (n = 25) for 26 wk. Changes in plasma HDL cholesterol, LDL cholesterol, and triglyceride concentrations did not differ between the 2 groups at 26 wk. The adjusted plasma total cholesterol concentration at 26 wk was 0.45 mmol/L higher in the flaxseed oil group (5.43 +/- 0.03 mmol/L) compared with the olive oil group (5.17 +/- 0.07 mmol/L) (P = 0.026). ALA did not affect LDL, HDL, or IDL particle size; however, the concentrations of the large, less atherogenic LDL1 (P = 0.058) and LDL2 (P = 0.083) subfractions tended to be greater in the ALA group. In conclusion, ALA does not decrease CVD risk by altering lipoprotein particle size or plasma lipoprotein concentrations.     

Lipoprotein oxidation mediated by J774 murine macrophages is inhibited by individual red wine polyphenols but not by ethanol

The in vitro capacities of individual polyphenols in red wine to inhibit the cell-mediated oxidation of lipoproteins and their effects on cell viability were determined. LDL and HDL were incubated with J774.A1 macrophages and 2 and 4 micro mol/L copper, respectively, in the absence and presence of polyphenols in ethanol at concentrations found in red wine. A mixture of polyphenols in amounts found in red wine equivalent to 0.2 g/L ethanol and 0.05 g/L ethanol inhibited thiobarbituric acid-reactive substance production from LDL by 91.7 and 45.9%, respectively, compared with ethanol controls (P < 0.01). HDL oxidation was inhibited 85 and 82.4% by the polyphenols at 0.2 and 0.05 g/L ethanol (P < 0.01). The effects of the polyphenol mixture on LDL oxidation were confirmed by measuring production of conjugated dienes and lipid peroxides, and trinitrobenzene sulfonic acid reactivity. Catechin at the concentration found in red wine (1.32 micro mol/L) at an ethanol concentration equivalent to 0.2 g/L inhibited LDL oxidation by 83.2%, while epicatechin (0.56 micro mol/L) and gallic acid (1.02 micro mol/L) inhibited by 60.6 and 26.9%, respectively (P < 0.05). At 1 micro mol/L, LDL oxidation was inhibited by epicatechin, catechin and quercetin by 86.2, 79.9 and 69.4%, respectively (P < 0.05). Incubation of macrophages with ethanol alone and with polyphenols in ethanol did not affect cell viability. Our results indicate that catechin and epicatechin are the major contributors to the antioxidant activity of red wine.     

Effect of phenol-rich extra virgin olive oil on markers of oxidation in healthy volunteers

OBJECTIVE: We studied whether consumption of phenol-rich extra virgin olive oil affects the susceptibility of low density lipoproteins (LDL) to oxidation and other markers of oxidation in humans. DESIGN: Randomized cross-over intervention trial, stratified according to sex, age and energy intake. SETTING: Division of Human Nutrition and Epidemiology, Wageningen University, The Netherlands. SUBJECTS: Forty-six healthy men and women completed the study. INTERVENTION: Subjects consumed two diets supplying 69 g per day of extra virgin olive oil either rich or poor in phenols for 3 weeks each. The mean difference in phenol intake between the treatments was 18 mg per day. Vitamin E intake was low during the whole study. Fasting blood samples were taken twice at the end of each period. RESULTS: Resistance of LDL and high density lipoprotein (HDL) to oxidation was not affected by treatment. The mean lag time of copper-induced formation of conjugated dienes was 1.6 min shorter in LDL and 0.4 min longer in HDL after the high phenol diet. Other markers of antioxidant capacity in plasma were also not affected: mean lipid hydroperoxides were 0.07 micromol/l higher, mean malondialdehydes were 0.001 micromol/l higher, mean protein carbonyls were 0.001 nmol/mg protein lower, and the mean ferric reducing ability of plasma (FRAP) was 0.006 mmol/l higher after the high phenol diet. All 95% confidence intervals enclosed zero. Serum cholesterol concentrations were not affected by the treatment. CONCLUSION: Consumption of 18 mg per day of phenols from extra virgin olive oil for 3 weeks did not affect LDL or HDL oxidation or other markers of antioxidant capacity in fasting plasma samples.     

Variability in alpha-tocopherol antioxidant activity in the core and surface layers of low- and high-density lipoproteins.

The effect of alpha-tocopherol enrichment of low- and high-density lipoproteins on Cu(2+)-catalyzed lipid peroxidation in the hydrophobic core and in the hydrophilic envelope of lipoproteins was investigated by using two pyrene derivatives, namely, cholesteryl pyrenyl hexanoate (P6Chol) and pyrene dodecanoyl sulfatide (P12CS). The progressive decrease in fluorescence of P6Chol was used to monitor lipid peroxidation in the core of LDL and HDL, whereas that of P12CS was used to follow lipid peroxidation in the envelope of both lipoproteins. alpha-Tocopherol enrichment of LDL and HDL was obtained by incubating blood plasma at 37 degrees C with different concentrations of the vitamin (25-500 microM) before lipoprotein separation. The incorporation of alpha-tocopherol in LDL and HDL presents a progressive, time-dependent increase up to 200 microM alpha-tocopherol, then a plateau up to 500 microM. In the envelopes, the added tocopherol causes a great decrease in the rate of peroxidation and a dramatic increase in the latency phase in both lipoproteins. In the cores the lengthening of latency phase resulting from alpha-tocopherol enrichment was by far greater in LDL than in HDL, and the decrease in the rate of peroxidation in both lipoproteins was less than in the envelopes.     

Multifunctional in vitro antioxidant evaluation of strawberry (Fragaria virginiana Dutch.)

Berries have shown the highest potential antioxidant activity among fruits using a unique antioxidant method. The antioxidant activity of aqueous-organic extracts of strawberry (Fragaria virginiana Dutch., var. camarosa) were determined using three methods: 2,2-diphenyl-1-picrylhydrazyl (DPPH*) radical scavenging, antioxidant ferric-reducing power (FRAP), and inhibition of Cu(II)-catalysed in vitro human low-density lipoprotein (LDL) oxidation. A serving (100 g) of strawberry had a DPPH* activity equivalent to that of 183 mg vitamin C and to that of 483 mg vitamin E. In addition, strawberry extracts showed high efficiency in the FRAP assay and in the in vitro inhibition of LDL oxidation. Regression linear analysis between radical scavenging activity (EC50 parameter) and total phenol content from strawberry extracts gave a statistically significant correlation (r = 0.984, P < 0.05). In conclusion, strawberry showed significant antioxidant capacity in both aqueous and lipophilic models.     

A mixed fruit and vegetable concentrate increases plasma antioxidant vitamins and folate and lowers plasma homocysteine in men

Fruit and vegetable consumption is inversely associated with coronary heart disease (CHD) risk. The aim of the present study was to determine the effect of supplementation with dehydrated juice concentrates from mixed fruit and vegetables on selected plasma vitamins and antioxidant status. We assessed CHD risk by measuring the concentrations of homocysteine, lipids, lipoproteins, glucose and insulin. Men were recruited to participate in a randomized double-blind, crossover trial with 2 periods of 6 wk, separated by a 3-wk wash-out period. Supplementation with the encapsulated mixed extract (Juice Plus) was compared with physically similar placebo capsules. Thirty-two men (13 smokers, 19 nonsmokers) completed the study with a mean compliance of 88%. Compared with placebo, supplementation increased the concentrations of plasma beta-carotene (0.24 +/- 0.15 vs. 1.12 +/- 0.70 micro mol/L; mean +/- SD; P < 0.0001), retinol (1.87 +/- 0.33 vs. 2.00 +/- 0.43 micro mol/L; P < 0.05), alpha-tocopherol (16.8 +/- 7.3 vs. 19.3 +/- 6.8 micro mol/L; P < 0.01), ascorbic acid (72.1 +/- 19.4 vs. 84.1 +/- 13.5 micro mol/L; P < 0.002) and folic acid (24.5 +/- 10.0 vs. 44.9 +/- 16.9 nmol/L; P < 0.0001). Plasma homocysteine was reduced (8.2 +/- 1.5 vs. 7.6 +/- 1.1; P < 0.05) and inversely related (r = -0.40, P < 0.001) with serum folate concentrations. Plasma vitamin C was positively correlated with the resistance of LDL to oxidation (r = 0.26, P < 0.05) and the plasma ferric reducing/antioxidant power (FRAP) tended to be greater after supplementation than after the placebo period (1125.5 +/- 144.1 vs. 1180.3 +/- 158.1 micro mol/L; P < 0.065). Plasma glucose, insulin and lipid concentrations were unaffected. Responses of smokers and nonsmokers did not differ. In the absence of dietary modification, supplementation with a fruit and vegetable concentrate produced responses consistent with a reduction in CHD risk.     

Human adrenal glands secrete vitamin C in response to adrenocorticotrophic hormone

BACKGROUND: When vitamin C intake is from foods, fasting plasma concentrations do not exceed 80 micromol/L. We postulated that such tight control permits a paracrine function of vitamin C. OBJECTIVE: The purpose of this study was to determine whether paracrine secretion of vitamin C from the adrenal glands occurs. DESIGN: During diagnostic evaluation of 26 patients with hyperaldosteronism, we administered adrenocorticotrophic hormone intravenously and measured vitamin C and cortisol in adrenal and peripheral veins. RESULTS: Adrenal vein vitamin C concentrations increased in all cases and reached a peak of 176 +/- 71 micromol/L at 1-4 min, whereas the corresponding peripheral vein vitamin C concentrations were 35 +/- 15 micromol/L (P<0.0001). Mean adrenal vein vitamin C increased from 39 +/- 15 micromol/L at 0 min, rose to 162 +/- 101 micromol/L at 2 min, and returned to 55 +/- 16 micromol/L at 15 min. Adrenal vein vitamin C release preceded the release of adrenal vein cortisol, which increased from 1923 +/- 2806 nmol/L at 0 min to 27 191 +/- 16 161 nmol/L at 15 min (P<0.0001). Peripheral plasma cortisol increased from 250 +/- 119 nmol/L at 0 min to 506 +/- 189 nmol/L at 15 min (P<0.0001). CONCLUSIONS: Adrenocorticotrophic hormone stimulation increases adrenal vein but not peripheral vein vitamin C concentrations. These data are the first in humans showing that hormone-regulated vitamin secretion occurs and that adrenal vitamin C paracrine secretion is part of the stress response. Tight control of peripheral vitamin C concentration is permissive of higher local concentrations that may have paracrine functions.