Whole plasma oxidation assay as a measure of lipoprotein oxidizability

Kontush, Anatol; Spranger, T.; Reich, Axel; Djahansouzi, S.; Karten, Barbara; Braesen, Jan Hinrich; Finckh, Barbara; Kohlschütter, Alfried; Beisiegel, Ulrike

doi:10.1002/biof.5520060203

Cited by 33 publications

(18 citation statements)

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“…The effect of ingestion of green tea on plasma oxidation ex vivo was determined using the method of Kontush et al [26]. Plasma samples were incubated at 37°C in the presence of PBS-EDTA buffer containing 50 µM CuSO4, and conjugated dienes production was measured by the changes of the absorbance at 234 nm.…”

Section: Effect Of Catechins On Ldl Oxidation In Vitromentioning

confidence: 99%

Effects of Catechins and Ground Green Tea Drinking on the Susceptibility of Plasma and LDL to the Oxidation In Vitro and Ex Vivo

Gomikawa¹,

Ishikawa²

2002

J. Clin. Biochem. Nutr.

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SummaryWe investigated the effects of catechins and ground green tea drinking on the susceptibilities of human low density lipoprotein (LDL) and plasma to the oxidation by CuSO4 in vitro and ex vivo. The effects of (-)epicatechin (EC) and (-)epigallocatechin gallate (EGCg) on LDL oxidation in vitro were estimated by measuring the lag time of conjugated diene formation, lipid hydroperoxide and agarose gel electrophoretic mobility. EC and EGCg had antioxidant activities for LDL oxidation, and the antioxidant activity of EGCg to LDL oxidation was stronger than that of EC in vitro. In an ex vivo study, we investigated the effects of ground green tea drinking on LDL and plasma oxidation after the consumption of a cup of green tea (5 g). Plasma total catechin contents were increased at 1 and 3 h after the oral ingestion, and we detected (-)epigallocatechin (EGC), EC, EGCg and (+)catechin (CA) in plasma. The lag time of conjugated diene formation of plasma was increased in all subjects after consumption of green tea (p<0.028), but the susceptibility of LDL oxidation ex vivo was different among individual subjects. Although the f3-carotene, o.-tocopherol and uric acid contents in plasma did not change after green tea consumption, ascorbic acid contents in plasma increased at 3 h after green tea ingestion. These findings suggest that catechins suppressed the susceptibilities of human LDL to oxidation by CuSO4 in vitro and plasma oxidation ex vivo after ground green tea ingestion.

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Section: Effect Of Catechins On Ldl Oxidation In Vitromentioning

confidence: 99%

Effects of Catechins and Ground Green Tea Drinking on the Susceptibility of Plasma and LDL to the Oxidation In Vitro and Ex Vivo

Gomikawa¹,

Ishikawa²

2002

J. Clin. Biochem. Nutr.

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“…On the other hand, the Cu 2ϩ -induced model of lipoprotein oxidation offers practical advantages with respect to simplicity, good reproducibility, and efficient sample processing (19,35).…”

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confidence: 99%

Neonatal Blood Plasma Is Less Susceptible to Oxidation Than Adult Plasma Owing to Its Higher Content of Bilirubin and Lower Content of Oxidizable Fatty Acids

et al. 2003

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Newborn infants are susceptible to a range of problems attributed to excessive production of free radicals. Because of a higher content of antioxidants, above all bilirubin, and a lower content of oxidizable lipids, newborn plasma should be better protected against oxidation than adult plasma. To test this hypothesis, we measured the susceptibility of plasma to in vitro oxidation in microsamples (7 L) from 57 healthy newborns and 18 adults. Heparin plasma was diluted 150-fold and oxidized by 50 M Cu 2ϩ . Oxidation was monitored as an increase in sample absorbance at 234 nm. Plasma oxidizability was found to be significantly lower in newborns than in adults. Accordingly, the level of bilirubin, an important antioxidant, was significantly higher, and the level of polyunsaturated fatty acids, a major substrate of lipid peroxidation, was significantly lower in newborn plasma. In addition, plasma oxidizability correlated positively with the level of polyunsaturated fatty acids and negatively with that of bilirubin. These data indicate that plasma is better protected against oxidative stress in newborns than in adults, owing to its higher content of antioxidants like bilirubin and its lower content of oxidizable lipids. Abbreviations MUFA, monounsaturated fatty acid PUFA, polyunsaturated fatty acid SFA, saturated fatty acid TRAP, total radical-trapping ability of plasma Newborn infants, especially premature ones, are susceptible to a wide range of problems related to an excessive production of free radicals (1, 2). In utero, fetal organs are exposed to relatively hypoxic tensions, which increase abruptly after birth (3). This transition may cause oxidative injury.In healthy humans, a balance exists between oxygen-derived free radical production and their removal by antioxidants (4). In preterm infants, inadequate antioxidant defenses may contribute to the pathogenesis of some complications of prematurity. Oxygen radical injury may be a common pathogenic mechanism in several neonatal diseases. For this reason, the term "oxygen radical disease of prematurity" was proposed for intracerebral hemorrhage, bronchopulmonary dysplasia, and retinopathy in premature neonates (1).Antioxidants are therefore of critical importance in protecting newborns against oxygen toxicity. Low plasma antioxidant activity at birth has been reported to be an independent risk factor for mortality in preterm babies (5). The measured and calculated TRAP was higher in newborns than in adults (6) and declined postnatally with increasing age (7). Accordingly, the susceptibility of cord plasma to in vitro oxidation by Cu 2ϩ was significantly lower in comparison with maternal plasma (8).Increased TRAP in newborns and the decreased susceptibility of neonatal plasma to oxidation are likely related to its higher content of two potent antioxidants, namely urate and, more importantly, bilirubin, which is highly increased in newborn plasma (7, 9 -11). Similarly, higher levels of selenium and other important antioxidants such as sulfhydryl groups have also bee...

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“…This method of lipoprotein oxidation in the arterial wall is therefore more relevant than the in vitro oxidation of single isolated lipoprotein such as LDL. Measurement of absorbance at 234 nm reflects the lipoprotein oxidation induced in vitro in whole plasma [18]. It is also known that dietary flavonoids reduce the oxidizability of LDL in vitro [38].…”

Section: Discussionmentioning

confidence: 99%

“…Plasma samples were diluted 100-fold with phosphate-buffered saline (PBS), pH 7.4, containing 0.16 M NaCl and oxidized in the presence of AAPH at 37°C. The change in sample absorbance was monitored spectrophotometrically at 234 nm at 30-min intervals for 20 hours [18]. The plasma oxidation was studied in the presence and absence of T. arjuna extracts as well as baicalein.…”

Section: Whole Plasma Oxidation Assaymentioning

confidence: 99%

“…To examine the mechanism underlying the known cardioprotective properties of T. arjuna and baicalein, in vitro studies of lipoprotein oxidation in whole human plasma were carried out. This seems to be a more relevant model of the lipoprotein oxidation in the arterial wall than that of a single isolated lipoprotein such as LDL [18]. To further reveal the more detailed mechanisms of baicalein responsible for the observed antioxidant effects, we allowed baicalein to react with some biologically relevant ROS as studied by different physicochemical methods such as spin trapping by electron spin resonance (ESR), and pulse radiolysis.…”

Section: Introductionmentioning

confidence: 99%

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Cellular Membrane Protection Against Reactive Oxygen Species by Terminalia Arjuna and Its Active Component Baicalein

Tilak

Devasagayam

Adhikari

et al. 2006

J. Clin. Biochem. Nutr.

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Summary Terminalia arjuna (T. arjuna) is an Indian medicinal plant credited with cardiotonic and cardioprotective properties; and baicalein is the active ingredient. To determine the possible mechanisms of protective action of T. arjuna and baicalein, we studied the reactions of T. arjuna extracts and baicalein with the biologically important superoxide (O2˙−) and singlet oxygen ( 1 O2) by measuring O2˙− or 1 O2 induced damage of lipids, lipid preoxidation, in the reaction mixture containing rat liver mitochondria and cardiac homogenate. Absorption of T. arjuna and baicalein in rat small intestine was also assessed. In addition, inhibitory effect of T. arjuna and baicalein on radiation-induced ROS generation in NIH3T3 cells and a cell-free chemical system, as well as on whole plasma oxidation were studied. Significant absorption of T. arjuna and baicalein from the intestine was observed. Baicalein is highly effective in inhibiting lipid peroxidation, reactive oxygen species generation, and plasma oxidation, even at low concentrations of 10-25 µM. To determine the possible mechanisms in this phenomena, we studied the reaction of T. arjuna extracts and baicalein with O2˙− and 1 O2 using electron spin resonance and found that T. arjuna extracts and *To whom correspondence should be addressed. Tel: +81-75-251-5503Fax: +81-75-252-3721 E-mail:toshi@koto.kpu-m.ac.jp Abbreviations: ˙OH, hydroxyl radical; µs, microseconds; 1 O2, singlet oxygen; 4-OH TEMP, 2,2,6,6-tetramethyl-4-hydroxypiperidine; 4-OH TEMPO˙, 2,2,6,6-tetramethyl-4-hydroxypiperidine-N-oxide radical; AAPH, 2.2'-azobis (2-amidinopropane) dihydrochloride; CD, conjugated diene; CO2˙−, formate radical anion; CO2, carbon dioxide; DCF, dichlorofluorescein; DCFH, 2',7'-dichlorofluorescein; DCFH/DA, 2',7'-dichlorodihydrofluorescein diacetate; DETAPAC, diethylenetriaminepentaacetic acid; DMEM, Dulbecco's modified eagle's medium; DMF, N,N'-dimethyl formamide; DMPO, 5,5-dimethyl-1-pyroline-N-oxide; eaq − , hydrated electrons; EDTA, ethylene diamine tetra acetic acid; ESR, electron spin resonance; H˙, hydrogen atom; H2O2, hydrogen peroxide; HCOO − , formate anion; IC50, concentration at which 50% inhibition is achieved; KSCN, potassium thiocyanate; LDL, low density lipoprotein; LOOH, lipid hydroperoxides; MeCN, acetonitrile; ms, milliseconds; N2, nitrogen; N2O, nitrous oxide; ns, nanoseconds; O2˙−, superoxide radical; OH − , hydroxyl anion; PBS, phosphate-buffered saline; PUFA, polyunsaturated fatty acid; RB, Rose Bengal; RFI, relative fluorescence intensity; RNS, reactive nitrogen species; ROS, reactive oxygen species; SAQA, singlet oxygen quenching activity; SSA, superoxide scavenging activity; TBARS, thiobarbituric acid reactive substances; Trolox, 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid. . In conclusion, our results show that T. arjuna and baicalein are very potent in membrane protection against O2˙− and 1 O2 in vitro, and their ability to react with these species may explain previously observed effects of this plant and its active ingredient.

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Whole plasma oxidation assay as a measure of lipoprotein oxidizability

Cited by 33 publications

References 34 publications

Effects of Catechins and Ground Green Tea Drinking on the Susceptibility of Plasma and LDL to the Oxidation In Vitro and Ex Vivo

Effects of Catechins and Ground Green Tea Drinking on the Susceptibility of Plasma and LDL to the Oxidation In Vitro and Ex Vivo

Neonatal Blood Plasma Is Less Susceptible to Oxidation Than Adult Plasma Owing to Its Higher Content of Bilirubin and Lower Content of Oxidizable Fatty Acids

Cellular Membrane Protection Against Reactive Oxygen Species by Terminalia Arjuna and Its Active Component Baicalein

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