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Publications and source records attributed to D Bonnefont-Rousselot.
Oxidative stress is implicated in many pathological processes and results from a disruption of the prooxidant/antioxidant balance. This review will focus on noninvasive biomarkers of radical-induced damage in biological fluids and particularly in blood. Special attention will be addressed to new analytical methods for the measurement of radical-mediated alterations in the integrity of lipids, proteins and DNA.
This study was designed to evaluate the antioxidant effect of aminoguanidine toward human low-density lipoproteins (LDLs) initiated by oxygenated free radicals (*OH/O(2)*-) generated by gamma radiolysis. Initial radiolytic yields related to the markers of lipid peroxidation [i.e. decrease in endogenous alpha-tocopherol and beta-carotene, formation of thiobarbituric acid-reactive substances (TBARS) and conjugated dienes] were determined in 3 g liter(-1) LDLs (expressed as total LDL concentration) in the absence and presence of 10 different concentrations of aminoguanidine (from 0.04 to 5 mmol liter(-1)). Fluorescence and relative electrophoretic mobility of oxidized LDLs were also studied as markers that indirectly reflect the attack of the protein moiety of LDLs (namely apolipoprotein B). Our data clearly showed the inhibitory effect of aminoguanidine on lipid peroxidation induced in LDLs by *OH/O(2)*- in a concentration-dependent manner. This effect probably resulted from a scavenging activity of aminoguanidine toward *OH. In contrast, aminoguanidine did not appear to react significantly with O(2)*-, which resulted in a poor residual lipid peroxidation. Our data led us to determine an optimum [aminoguanidine]/[LDL] ratio ranging from 250 to 500 to obtain the best in vitro protection of LDLs under our experimental conditions. It is also of great interest that aminoguanidine was able to protect endogenous alpha-tocopherol and beta-carotene of LDLs upon *OH/O(2)*(-)-induced oxidation.
It has been shown that elevated extra- and intra-cellular glucose concentrations result in an oxidative stress, which is defined as an imbalance between prooxidants and antioxidants. Several mechanisms seem to be involved in the genesis of this oxidative stress, which has been reported both in experimental diabetes in animals and in type 1 and type 2 diabetic patients: glucose autoxidation, protein glycation and formation of advanced glycation endproducts, and the polyol pathway. Reciprocally, oxidative stress is involved in the origin of type 1 diabetes, especially via the apoptosis of pancreatic beta-cells, as well as insulin resistance in type 2 diabetes. Glucose control plays an important role in the prooxidant/antioxidant balance. Macromolecules such as molecules of extracellular matrix, lipoproteins and deoxyribonucleic acid are also damaged by free radicals in diabetes mellitus. A supplementation with antioxidants has been proposed as a complementary treatment, and some antidiabetic agents may by themselves have antioxidant properties independently of their role on glucose control. The aim of this paper was to review the consequences of the diabetic status on the oxidant/antioxidant balance.
The aim of our study was to determine the oxysterol formation in low density lipoproteins (LDLs) oxidized by defined oxygen free radicals (*OH/O2*-). This was compared to the oxysterol produced upon the classical copper oxidation procedure. The results showed a markedly lower formation of oxysterols induced by *OH/O2*- free radicals than by copper and thus suggested a poor ability of these radicals to initiate cholesterol oxidation in LDLs. Moreover, the molecular species of cholesteryl ester hydroperoxides produced by LDL copper oxidation seemed more labile than those formed upon *OH/O2*(-)-induced oxidation, probably due to their degradation by reaction with copper ions.
Susceptibility of lipoproteins to oxidation is partly determined by their content in endogenous antioxidants, but also by the polyunsaturated fatty acids (PUFA)/monounsaturated fatty acids (MUFA) ratio. The aim of our study was to enrich human high-density lipoproteins (HDLs) with dioleoylphosphatidylcholine (DOPC) in order to modify the PUFA/MUFA ratio while maintainig the alpha-tocopherol/PUFA ratio constant and to appreciate the consequences of this enrichment before and after copper-induced oxidation. The enrichment of HDLs with DOPC was obtained by incubation of these lipoproteins with DOPC liposomes and further reisolation of HDLs. The consequent 40% HDL enrichment in MUFA was concomitant with a 35% loss in PUFA (MUFA/PUFA ratio = 1.43). The enrichment of HDLs with DOPC led to a 40% decrease in alpha-tocopherol content, which kept a constant alpha-tocopherol/PUFA ratio. The DOPC-HDLs exhibited a lower oxidizability by copper than the nonenriched HDLs (NE-HDLs), as shown by their twofold longer lag phase and the threefold lower propagation rate. Moreover, DOPC-HDLs led to a six- to sevenfold lower production of hydroperoxide molecular species from phosphatidylcholine and cholesteryl esters than NE-HDLs after 24 h copper oxidation. With regard to the cholesterol effluxing capacity, copper oxidation of HDLs led to a decrease of this property. However, our results clearly showed that DOPC enrichment of HDLs allowed us to keep a better effluxing capacity than in NE-HDLs after 24 h oxidation (22.3% vs 17.4%, respectively). Since apo A-I was degraded as well in DOPC-HDLs as in NE-HDLs, the better effluxing capacity of DOPC-HDLs could not come from a preserved integrity of apo A-I. It could be partly related to the improved fluidity of oxidized DOPC-HDLs compared to oxidized NE-HDLs, as shown by electron spin resonance data (correlation-relaxation time at 24 degreesC = 2.20 ns vs 3.00 ns after 24 h oxidation, in DOPC-HDLs and in NE-HDLs, respectively). Besides, it could also be hypothesized that the sevenfold lower content of phosphatidylcholine hydroperoxides in DOPC-HDLs than in NE-HDLs after 24 h copper oxidation could be involved in the better ability of oxidized DOPC-HDLs to mobilize cellular cholesterol.
According to the oxidative hypothesis of atherosclerosis, a hyperoxidizability of lipoproteins could favor the development of the atherosclerotic process. Besides, it has been recently reported that models of elevated very-low-density-lipoprotein (VLDL) levels in rats resulted in an increased susceptibility of these VLDL to oxidation. Treatment with dexamethasone classically induces an increase in plasma VLDL concentration. The aim of our study was thus to assess the effects of a treatment with dexamethasone in rats on the susceptibility to copper oxidation, both on total plasma and on isolated lipoproteins. Male Sprague-Dawley rats aged three months were treated with a daily intraperitoneal injection of dexamethasone (1.5 mg per kg) for five days (DEX group), whereas control rats were fed ad libitum (AL group). In order to take into account the decrease of food intake induced by dexamethasone treatment, a group of pair-fed rats was constituted (PF group). These rats had the same food intake as rats of the DEX group and were treated with a daily isovolumic intraperitoneal injection of NaCl for 5 d. After 5 d treatment, rats were fasted overnight, then killed, and blood was collected on EDTA. Low-density lipoproteins (VLDL + LDL) and high-density lipoproteins (HDL) were isolated by ultracentrifugation. A copper oxidation was conducted both on total plasma and on isolated lipoproteins. As expected, after treatment with dexamethasone, plasma exhibited increased triglyceride and glucose levels. Similarly, VLDL + LDL of rats from the DEX group were enriched with triglycerides, when compared with VLDL + LDL of the other two groups of rats. Our major finding was a marked increase in the susceptibility of total plasma of the DEX group to copper oxidation, in comparison with the other two groups of rats. This oxidizability was assessed by the maximal level of oxidation products absorbing at 234 nm and classically considered to be conjugated dienes (7.46+/-0.70 micromol L(-1) in the DEX group vs. 3.36+/-0.40 and 2.05+/-0.60 micromol L(-1) in the AL and PF groups, respectively). Nevertheless, this higher oxidizability was not observed in the isolated lipoprotein fractions, as shown by the formation of lipid peroxidation products such as conjugated dienes, thiobarbituric-acid reactive substances, hydroperoxides, 7-ketocholesterol, and dienals. This is not in agreement with other models of hypertriglyceridemia that have been reported to induce a hyperoxidizability of lipoproteins in rats. Our results led us to hypothesize that other plasma components such as proteins could be involved in this susceptibility to oxidation. Indeed, the severe protein catabolism induced by dexamethasone treatment could support this hypothesis, by forming protein components that are more susceptible to oxidation, as shown by an increased carbonyl formation upon plasma copper oxidation.
The oxidative hypothesis of atherosclerosis classically implies a central role for low density lipoprotein (LDL) oxidation. However, new antiatherogenic properties have been recognized for high density lipoproteins (HDL), apart from their ability to reverse cholesterol transport. Indeed, native HDL could protect LDL from oxidation, thereby minimizing the deleterious consequences of this process. Several mechanisms have been suggested to explain this protective role. Two HDL-associated enzymes, paraoxonase and PAF-acetylhydrolase, detoxify oxidized phospholipids produced by lipid peroxidation. In addition, HDL could reduce hydroperoxides to their corresponding hydroxides. It has also been suggested that HDL could inhibit oxidized LDL-induced transduction signals. However, in vivo HDL oxidation in the subendothelial space would favor the atherosclerotic process. Indeed, atherogenic properties of these oxidized HDL partly result from some loss of their cholesterol effluxing capacity and from an inactivation of the lecithin-cholesterol acyltransferase, which is a HDL-associated enzyme involved in reverse cholesterol transport. Finally, oxidized HDL could induce cholesterol accumulation in macrophages. Further in-depth investigation is needed to assess these antagonistic effects and their consequences for the atherosclerotic process.
This study was designed to evaluate the antioxidant effect of probucol on peroxidation of low-density lipoproteins (LDLs) initiated by oxygenated free radicals (O2*-) and ethanol-derived peroxyl radicals (RO2*) generated by gamma radiolysis. Initial radiolytic yields related to the markers of lipid peroxidation [i.e. decrease in endogenous alpha-tocopherol, formation of thiobarbituric acid-reactive substances (TBARS) and conjugated dienes] were determined as a function of LDL concentration (1.5 and 3 g l(-1), expressed as total LDL) and in the absence or the presence of probucol at different concentrations (2.3 x 10(-6), 3.5 x 10(-6), 9 x 10(-6) and 20.5 x 10(-6) mol l(-1)). Our results showed that probucol was able to decrease not only the yields of TBARS and conjugated dienes but also the levels of these peroxidation products obtained at high doses (2500 Gy) compared to LDLs without probucol. Under our conditions, probucol displayed an optimal antioxidant effect for an initial concentration in LDLs equivalent to 15 probucol molecules per LDL particle, which corresponded to a pharmacologically relevant concentration of probucol. Moreover, our data showed that probucol was unable to react with RO2* and thus did not protect LDL vitamin E from free radical attack. In addition, the scavenging capacity of probucol on O2*- appeared to be very poor, and probucol more likely reacted with LDL intermediate radical products. Finally, a very significant steady-state level of probucol remained in LDLs at high doses (up to 2500 Gy), equivalent to at least 40% of the initial concentration of probucol. This addressed the question of a mechanism for regeneration of probucol in LDLs. Our results as a whole suggested that the antioxidant effect of probucol in vivo could not be explained by its scavenging capacity with regard to RO2*/O2*- free radicals.
BACKGROUND AND AIM: The aim of the study was to describe qualitative and/or quantitative modifications of lipoproteins, including their fatty acid composition, in obese patients during a hypocaloric diet, and determine whether the variations observed paralleled modifications of alpha-tocopherol concentration in adipose tissue and blood. METHODS AND RESULTS: 15 healthy, obese volunteers (5 males, 10 females; age: 32-69 yr; BMI: 28.4-60.5 kg/m2) were given a 3-week hypocaloric diet (3.9 MJ (941 kcal)). Adipose tissue and blood samples were taken at the beginning and at the end of this period. At baseline and after 3 weeks measurements were made for alpha-tocopherol and fatty acids in total serum, lipoproteins and adipose tissue; thiobarbituric acid-reactive substances (TBARS) in serum. A significant drop in cholesterol-rich particles (LDL and HDL) was observed, in parallel to a significant enrichment of n-6 polyunsaturated fatty acids (PUFA) at the expense of both saturated and monounsaturated fatty acids in serum. A drop in alpha-tocopherol concentration (expressed as mumol alpha-tocopherol/g lipid) in serum and lipoprotein fractions paralleled the decrease in cholesterol-rich lipoproteins. CONCLUSIONS: Our results suggest that a hypocaloric diet not only decreases cholesterol-rich particle levels in serum, but also leads to a significant modification of fatty acid composition which may reflect improvement of insulin sensitivity. We did not observe any modification in adipose tissue after diet with regard to both alpha-tocopherol and fatty acid concentrations. Despite a drop in alpha-tocopherol concentration and an increase in n-6 PUFA content in serum, we did not find any enhancement of serum lipid peroxidation level evaluated by the thiobarbituric acid-reactive substance (TBARS) assay. If we assume that dietary intakes of alpha-tocopherol were not modified during this diet, it can be supposed that adipose tissue released alpha-tocopherol without any specific regulation, in parallel to the release of fatty acids.
In this study, human low-density lipoprotein (LDL) vesicles were irradiated with 73 MeV protons (LET of 1 keV/microm) and 11.4 MeV/nucleon argon ions (LET of 1.52 MeV/pm) and the effectiveness of charged particles in initiating peroxidation of LDLs was investigated. The LDL suspension (6 g/l) was exposed to protons and to argon ions in a dose range of 24 Gy to 2.4 kGy. Irradiations were carried out at the synchrocyclotron at the CPO and at the UNILAC of the GSI. After irradiation three chemical assays were used to study the progression of peroxidation of LDLs: the formation of conjugated dienes, the formation of thiobarbituric acid-reactive substances (TBARS) and the increase in the relative electrophoretic mobility of the LDLs. The results were compared with those obtained after gamma irradiation. For protons the yields of the peroxidation products were 10 times lower than after gamma irradiation. However, for doses below 200 Gy, protons appeared to be more effective than gamma rays in damaging the protein moiety, as deduced from the observed increase in the relative electrophoretic mobility of the LDLs. The irradiation with argon ions led to a negligible formation of peroxidation products, but an increase in the relative electrophoretic mobility of the LDLs was observed. The results are indicative of a lower yield of lipid peroxidation after irradiation with high-LET particles. In contrast, protons and argon ions appear to be more effective in inducing bulk protein and phospholipid damage than gamma rays.
Oxidation of low density lipoprotein is involved in the pathogenesis of atherosclerosis. Epidemiological studies suggest a negative correlation between the occurrence of cardiovascular diseases and blood concentrations of lipophilic antioxidants such as vitamin A and E and beta-carotene. Trace elements such as selenium, zinc and copper are involved in the activity of antioxidant enzymes: glutathione peroxidase and superoxide dismutase. The aim of this work was to determine the antioxidant and trace elements status of patients with very severe hypercholesterolemia and who were treated by dextran sulphate low density lipoprotein apheresis, in comparison with two control populations: one constituted by normocholesterolemic subjects and the other by hypercholesterolemic patients before treatment. Our results showed that, as compared with normocholesterolemic subjects, patients treated by LDL-apheresis were not deficient in vitamin E, beta-carotene and copper but had low plasma levels of selenium, zinc and vitamin A. The low selenium and vitamin A levels were due to the treatment by LDL-apheresis by itself, while the hypercholesterolemia of these patients might have provoked the low plasma levels of zinc. This study pointed out the interest of a supplement of selenium, zinc and vitamin A in patients treated by LDL-apheresis.
Series of indole-2-carboxamide and cycloalkeno[1,2-b]indole derivatives were synthesized and evaluated in order to determine the necessary structural requirements for a high inhibition of human LDL copper-induced peroxidation. Various modulations were systematically performed on the indole and cycloalkeno[1,2-b]indole nuclei as well as on the carboxamide moiety. The best compounds (3c, 3e, 7c, 7f, 7h, 7g, and 7o) are between 5 and 30 times more active than probucol itself. Two of these compounds (3c and 7o) were selected for complementary in vitro and in vivo investigations, which have shown additional properties of interest for the treatment and the prevention of atherosclerosis injuries. Compound 3c was found to have some antiinflammatory properties while compound 7o was proved to protect endothelial cells from the direct cytotoxicity of oxidized LDL with some additional calcium channel blocking properties.
The aim of this work was to compare the behaviour of HDL oxidised by .OH or .OH/O2.- free radicals produced by gamma radiolysis in the absence or in the presence of LDL at the same concentration of 3 g x l(-1), in order to specify the possibility of reciprocal protection of HDL and LDL towards lipid peroxidation. This oxidation was quantitatively evaluated by the decrease of endogenous alpha-tocopherol and the formation of oxidation products (thiobarbituric acid-reactive substances and conjugated dienes) and by the determination of initial radiation yields. Our results demonstrated that HDL could be protected by LDL against in vitro radical oxidation only in the presence of oxygen (action of .OH/O2.- free radicals). This observation addresses new questions about the interaction between HDL and LDL, especially the possibility of a reciprocal protection.
In the aim to study the effect of an in vitro enrichment of high-density lipoprotein (HDL) with alpha-tocopherol in alcoholic solution on a copper-induced peroxidation, we monitored several markers of lipid peroxidation (alpha-tocopherol consumption, formation of conjugated dienes and of fatty acid hydroperoxides, production of thiobarbituric acid-reactive substances) and the integrity of apolipoprotein A-I. High-density lipoproteins (1.063 < d < 1.21) with a mean of 0.58 alpha-tocopherol molecules per HDL particle were enriched with alpha-tocopherol in alcoholic solution to obtain an average of 3.7 and 21 alpha-tocopherol molecules per HDL particle. HDL oxidation with 5 microM CuSO4 at 37 degrees C resulted in the total disappearance of endogenous alpha-tocopherol after 2 h, but after 24 h about 19% of alpha-tocopherol remained in the most enriched HDL. In agreement with the tocopherol-mediated peroxidation, the formation of conjugated dienes and of fatty acid hydroperoxides was very fast and increased with alpha-tocopherol concentration, whereas TBARS production decreased. These results showed that alpha-tocopherol enrichment stabilized the production of hydroperoxides in HDL and decreased the formation of secondary oxidation products. These latter products are known for deleterious effects towards apolipoproteins. This could explain why we observed that the apolipoprotein A-I of the most enriched HDL was only slightly altered after incubation with CuSO4.
BACKGROUND: An oxidative stress has been reported in patients with chronic renal failure (CRF) treated by haemodialysis. To our knowledge, only scant information is available concerning CRF patients treated by continuous ambulatory peritoneal dialysis (CAPD) with regard to their redox and nutritional status. METHODS: The oxidative stress and the biological nutritional status were evaluated in 20 elderly CRF patients treated by CAPD, compared with a control group of 30 elderly non-CRF patients. Plasma peroxidation products were assayed as thiobarbituric acid-reactive substances (TBARS), and two enzymatic antioxidant systems were determined: erythrocyte superoxide dismutase (SOD), glutathione peroxidase activity in plasma (P-GSH-Px) and in erythrocytes (E-GSH-Px). Selenium, vitamin E, beta-carotene and vitamin A were evaluated as plasma non-enzymatic antioxidants. Nutritional status and iron status were assessed by determining serum albumin, prealbumin, iron, ferritin and transferrin concentrations. RESULTS: Plasma TBARS concentration was high in both groups (CAPD: 1.37 +/- 0.06 mumol/l versus non-CRF: 1.41 +/- 0.06 mumol/l; P = NS), compared with usual values (0.60 to 1.20 mumol/l), on account of the patients' ages. SOD and E-GSH-Px activities were normal in both groups. A significant lowering in P-GSH-Px activity was observed only in CAPD patients (211 +/- 14 U/l, usual values: 480 to 650 U/l). Plasma selenium concentration, decreased in both groups, was significantly lower in CAPD than in non-CRF patients (P < 0.01). Plasma vitamin E, beta-carotene and vitamin A concentrations were significantly enhanced only in CAPD patients (P < 0.0001, P < 0.005 and P < 0.0001, respectively. Biological nutritional markers were similar in both groups and within usual values. CONCLUSIONS: This study demonstrated the existence of an oxidative stress in CAPD-treated elderly CRF patients, evidenced by a decrease in plasma selenium levels and in P-GSH-Px activity. However, plasma TBARS were not higher in CAPD patients than in age-matched non-CRF control subjects, probably on account on the patients' ages.
The aim of this work was to specify the mechanisms involved in the radical oxidation of human high-density lipoprotein (HDL) and to compare these mechanisms with those described previously for the oxidation of low-density lipoprotein (LDL) under the same experimental conditions (Bonnefont-Rousselot et al., Radiat, Res. 134, 271-282, 1993). The oxidation of HDL, initiated by .OH or .OH/O(.-)2 free radicals from gamma radiolysis of water, was evaluated as a function of increasing radiation dose by analyzing quantitatively the decrease of endogenous alpha-tocopherol and the formation of oxidation products (thiobarbituric acid-reactive substances and conjugated dienes). All qualitative conclusions were supported by quantitative data (radiation yields and concentrations of the oxidation markers at high radiation doses) and by the mechanisms of the kinetics, .OH free radicals in the absence of oxygen were less efficient in initiating HDL oxidation than in the presence of oxygen (action of .OH/O(.-)2 free radicals), which was in agreement with the enhancement of the action of .OH free radicals by oxygen. The remaining significant level of vitamin E in HDLs at high radiation doses in the absence of oxygen could be explained by a regeneration of vitamin E by an oxidation product that was able to reduce the alpha-tocopheroxyl radical. The yields related to the decrease in the vitamin E content of HDLs after exposure to radiation with .OH or .OH/O(.-)2 free radicals were slightly higher than those obtained previously in LDLs under similar experimental conditions. Moreover, in the presence of oxygen, .OH free radicals led to a lower formation of thiobarbituric acid-reactive substances in HDLs than in LDLs. Such discrepancies in the behavior of these two lipoprotein fractions could be related to the differences in the chemical composition of HDLs and LDLs.
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Oxidation of low density lipoprotein is involved in the pathogenesis of atherosclerosis. Epidemiological studies suggest a negative correlation between the occurrence of cardiovascular diseases and blood concentrations of lipophilic antioxidants such as vitamins A and E and beta-carotene. Trace elements, such as selenium, zinc, and copper, are involved in the activity of the antioxidant enzymes glutathione peroxidase and superoxide dismutase. The aim of this study was to determine the antioxidant and trace element status of patients with severe hypercholesterolemia who had been treated with dextran-sulphate low-density lipoprotein apheresis in comparison with two control populations, normocholesterolemic subjects and untreated hypercholesterolemic patients. Our results showed that, patients treated with LDL apheresis, compared with normocholesteromic subjects, were not deficient in vitamin E, beta-carotene, and copper, but had lower plasma levels of selenium, zinc, and vitamin A. The low selenium and vitamin A levels were due to the LDL-apheresis treatment, and the hypercholesterolemia might have provoked the low plasma levels of zinc. The study pointed out the potential benefits of supplemental selenium, zinc, and vitamin A in patients being treated with LDL apheresis.