Lipid Peroxidation Products In Erythrocytes Research Articles

EFFECT AND KEY MECHANISMS OF ACTION OF PHOTOBIOMODULATION USING DEFOCUSED HIGH-POWER DIODE LASER DURING SKIN REPAIR

<h3>Objectives</h3> To evaluate the effect of different photobiomodulation therapy (PBMT) protocols in skin wound healing. <h3>Study Design</h3> 105 male Wistar rats were divided into 5 groups: negative control (<i>n</i> = 5; with no injury and without PBMT); sham (with injury and without PBM); 0.1 W laser; defocused high-power laser (DHPL) dual 1 W (DHPLD1W); DHPL dual 2 W (DHPLD2W). One full-thickness dorsal skin wound was created. PBMT groups received daily treatment. After 3, 5, 10, 14, and 21 days, euthanasia was performed for clinical, morphologic, oxidative stress (erythrocyte lipid peroxidation products (MDA), antioxidant enzymes superoxide dismutase (SOD) and glutathione (GSH)), and cytokine analyses (IL-1β, IL-10, and tumor necrosis factor alpha [TNF-α]). <h3>Results</h3> Clinically, at days 10 and 14, the DHPLD1W promoted faster tissue repair than the sham group (<i>P</i> < .05). Histopathologic evaluations showed that at day 3, inflammatory responses were lower in all PBMT groups (<i>P</i> < .05). At day 10, DHPLD1W exhibited lower inflammation and better reepithelization compared to other groups. Collagen analyses at day 10 revealed that all PBMTs promoted better maturation than the sham group (<i>P</i> < .05). MDA was lower and GSH and SOD were higher in the DHPLD1W and DHPLD2W groups and L0.1W led to an increase in IL-1β and TNF-α levels at day 3. <h3>Conclusions</h3> PBMT promoted skin wound healing, especially the DHPL1W protocol modulating the inflammatory process and oxidative stress and stimulating collagen maturation.

LED THERAPY IN SKIN REPAIR MODULATES CYTOKINE LEVELS AND OXIDATIVE STRESS

<h3>Objectives</h3> To evaluate the effect of light emitting diode (LED) therapy in skin wound healing. <h3>Study Design</h3> 55 male Wistar rats were divided into 3 groups: negative control (<i>n</i> = 5; with no injury and without treatment), sham (with injury and without treatment), and LED groups. One full-thickness dorsal skin wound was created. The LED group received daily treatment. After 3, 5, 10, 14, and 21 days, euthanasia was performed for clinical, morphologic, oxidative stress (erythrocyte lipid peroxidation products (MDA), antioxidant enzymes superoxide dismutase (SOD) and glutathione (GSH)), and cytokine analyses (IL-1β, IL-10, and tumor necrosis factor alpha [TNF-α]). <h3>Results</h3> Clinically, at day 10, the LED therapy promoted faster tissue repair than the sham group (<i>P</i> < .05). Histopathologic evaluations showed that in 3 and 10 days, the LED group exhibited more advanced resolution of inflammatory processes compared to the sham group. MDA was lower and GSH was higher in the LED group in all experimental periods. SOD was higher at 10 days and lower at 5 and 21 days (<i>P</i> < .01). On day 10 there was a significant difference in the levels of IL-1β, IL-10 (<i>P</i> < .05), and TNF-α (<i>P</i> < .01). <h3>Conclusions</h3> LED therapy has antioxidant action and modulates inflammatory cytokines but had limited clinical results with the parameters used.

Alleviation Effects of Diosmetin on H2O2-Induced Oxidative Damage in Human Erythrocytes

Free radicals (FRs) are formed in the high amounts result of the metabolic imbalance in cells and tissue. These radicals-induced oxidative damages constitute the basis of many diseases. Organisms have antioxidant defence systems (ADS) to eliminate the destructive effects of the oxidative damage. In addition to these antioxidant systems, dietary flavonoids have the antioxidant effect and the protective role against oxidative damage. In the present study, it was investigated whether a flavonoid derived diosmetin (10, 50, and 100 µM) have the elimination potential on hydrogen peroxide (H2O2)-induced oxidative damage in erythrocyte culture by using biomarkers such as lipid peroxidation (LP) level, catalase (CAT), total superoxide dismutase (SOD) activity and changes of SOD isozymes containing the manganese SOD (Mn SOD) and the cupper-zinc SOD (CuZn SOD). CAT, total SOD, Mn SOD and CuZn SOD activities showed a serious decline with H2O2 treatment, but diosmetin addition significantly increased their activities. While the H2O2 application critically increased LP products in erythrocytes, diosmetin considerably reduced these oxidative damage products. In conclusion, it has been determined that diosmetin can moderate oxidative damage in human erythrocytes by activating or protecting the ADS.

Probucol induces the generation of lipid peroxidation products in erythrocytes and plasma of male cynomolgus macaques.

We previously reported that probucol, a lipid lowering agent, protected mice from malaria infection via depletion in plasma α-tocopherol. The antioxidant α-tocopherol in host circulation is necessary for the malaria parasites to protect themselves from oxidative stress in erythrocytes where high amounts of reactive oxygen species are generated. To assess the potential for the clinical application of probucol as an anti-malarial therapy, it was necessary to determine the effects of probucol by using primate experiments. Here we verified that probucol induces an α-tocopherol decrement in cynomolgus macaque erythrocytes and plasma. After 2 weeks of probucol administration at doses of 200 or 400 mg/kg/day, the α-tocopherol contents in erythrocytes tended to decrease. The contents of hydroxyoctadecadienoic acids and 7β-hydroxycholesterol, peroxidation products derived from linoleic acid and cholesterol, respectively, increased in erythrocytes. On the other hand, plasma α-tocopherol concentration showed a marginal decrement. Plasma lipid peroxidation products were transiently increased in the early stages of probucol administration. No adverse effects were observed throughout the experiment, although the dosage of probucol was higher than the clinical maximum dosage. Considering that malaria proliferates in erythrocytes, probucol-induced disruption of redox homeostasis in erythrocytes could be effective in the inhibition of parasite proliferation.

EFFECT OF L-ARGININE - NO ON PROOXIDANT-ANTIOXIDANT BALANCE IN ERYTHROCYTES OF RATS UNDER ALCOHOL INTOXICATION.

It was shown changes in the activity of antioxidant enzymes (superoxide dismutase, catalase) and NO-synthase (NOS), in the content. of stable metabolic products of nitric oxide and levels of lipid peroxidation products in erythrocytes of rats under alcoholic intoxication. It was shown that animals with alcohol intoxication under of the admission of the main substrate NOS - L-arginine activity of antioxidant protection enzymes was increased in twice on the fond of TBA-positive products decrease contents. Established in hemolisate red blood cells in rats with alcohol inrotoxication value of total NOS activity decreases by 65% compared to control. Not selective inhibitor Nto-nitro-L-arginine methyl ester (L-NAME), which is a structural analog of L-arginine, reduced output level of total NOS activity by 23.4% in the control and 25% under conditions of pathology. The consumption of rats L-arginine NOS total activity increased in the two study groups. The results testify that L-arginine has antioxidant properties, whereas L-NAME exerts a slightly stabilizing influence.

Перекисне окислення білків та ліпідів при недиференційованій дисплазії сполучної тканини у дітей

Aim: The study of lipid and protein peroxidation in children with undifferentiated connective tissue dysplasia. Materials and Methods: There were studied the state of lipid and protein peroxidation on 63 children (33 children with undifferentiated connective tissue dysplasia (UDCTD) and 30 children without (UDCTD)) by spectrofotometrie (the change of index and the end product of lipid peroxide modification — malondialdehyde (MDA) in blood plasma and erythrocyte membranes, the process of peroxidation of proteins (PSP) — the content of the final products of oxidative modification — 2,4 dinitrophenylhydrazine blood plasma, the activity of the antioxidant defense enzymes — catalase (CT) and superoxide dismutase (SOD) activity in membrane structures and plasma) and biochemical methods (level of free cholesterol). Results: we established a significant increase in venous blood plasma products of lipid peroxidation, an increase of lipid peroxidation products in erythrocytes and an imbalance in the antioxidant defense system in children with UDCTD compared with a group of children without UDCTD. Conclusion: there were observed changes in the peroxidation of plasma proteins in venous blood (2,4 dinitrophenylhydrazine in apoproteids of lipoproteins low and very low density), the index of peroxide modification of lipoproteins and lipid membranes of red blood cells, an imbalance of antioxidant enzymes (SOD and CT), which the extent is correlated with the degree of phenotypic disorders in UDCTD in adolescents with UDCTD. Key words: children, undifferentiated connective tissue dysplasia, lipid peroxidation, protein peroxidation, antioxidant enzymes.

Prognostic biochemical markers of thrombolytic therapy in ischemic stroke

To assess prognostic value of endothelial dysfunction, endogenous intoxication and oxidative stress in the acute period of ischemic stroke (II) at the stage of making a decision on using thrombolytic therapy (TLT) as well during the follow-up in order to personalize treatment. The study included 57 patients stratified into control group 1 (n=29) and TLT group (n=28). Contents of C-reactive protein (Nsrb), matrixmetalloproteinase-9 (MMP-9) erythrocyte lipid peroxidation products were measured in the blood of patients. Clinical outcomes were evaluated with the modified Rankin scale and the Rivermead Mobility Index. Increased levels of Nsrb and MMP-9 were the biomarkers of poor TLT outcome. TLT has a positive effect on the basic patterns of IS pathogenesis and biochemical predictors allow to rationalize the treatment.

THE MAIN PARAMETERS OF THE ANTIOXIDANT SYSTEM IN BLOOD OF MALE AND FEMALE RABBITS IN AGE DYNAMICS

Abstract Antioxidant system (AOS) is the most important unit of the general biological defense sys-tem. In assessing its state the choice of adequate, stable and sensitive indicators is important to de-termine the products of lipid peroxidation (LPO) and antioxidant defense enzyme complex. Antioxi-dant Index (AOI) is based on detecting blood and erythrocyte lipid peroxidation products such as malondialdehyde, diene conjugates of polyunsaturated fatty acids, and assessing the activity of anti-oxidant enzymes (i.e. superoxide dismutase, catalase, glutathione peroxidase and glutathione reduc-tase) in erythrocyte membranes. The aim of this work was to evaluate the role of AOS in the mecha-nism of blood oxidative homeostasis in Soviet Chinchilla rabbits (males and females) during different periods of ontogeny. Studies were performed in bioclinic on 10 rabbits. Blood was sampled from animals at the age of 60, 120 and 180 days. In the blood plasma we determined the content of diene conjugates and malondialdehyde, antioxidants and low molecular weight -tocopherol and retinol. In hemolysates the activity of antioxidant enzymes, i.e. glutathione reductase, glutathione peroxidase, catalase, superoxide dismutase, was evaluated. For the first time for the biological evaluation of anti-oxidant system of rabbits a combined integrated AOI was used, which allows to reveal not only changes in the AOS of blood, but also to determine the proportion of its individual factors in the on-togenesis of rabbits. A comparison of the content of cholesterol, glucose and triglycerides in blood plasma showed the differences between male and female rabbits in lipid and energy metabolism. At the age of 180 days an increase in cholesterol, triglycerides and glucose in males was 11.6 %, 27.2 % (P  0.001) and 12.9 % (P  0.01), respectively, and in the females it was 14.7 %, 27.4 % (P  0.001) and 13.0 % (P  0.05), respectively. At the age of 120 and 180 days in the animals an insig-nificant reduction in all indicators was observed compared with the values obtained in the 60 day old rabbits. -Tocopherol content in males aged 120 and 180 days increased by 14.4 and 21.9 %, respec-tively, and the retinol level was 8.3 and 29.2 % higher compared to the initial (day 60) values. In females there was a significant increase in the concentration of -tocopherol by 14.9 and 18.9 %, re-spectively, and in retinol by 13.6 and 50.0 %, respectively, at days 120 and 180. Revealed changes in the overall and partial AOI in ontogeny indicate the initial activation of compensatory mechanism of antixidation in early ontogenesis and further decrease in a compensatory capacity of the antioxidant defense system in blood. Thus, the general index of the AOI in 4 month old rabbit males and fe-males increased by 12-20 % and 4-12 %, respectively. A positive index value indicates the relatively high antioxidant protection despite the trend to reducing activity of the enzyme protection. In the 6 month old animals the integrated combined AOI values were negative, being typical for the state of oxidative stress. In males these changes were more evident and reliable. Usage of AOI index may be informative in assessment of the state of protective systems both in farm and laboratory animals. Keywords: ontogeny, protection systems, rabbits, antioxidant system, AOS, blood antioxi-dant index, AOI.

Features and role of lipid peroxidation in patients with chronic hepatitis C (genotype 1b) before and after antiviral therapy

Aim. To estimate the levels of lipid peroxidation products in erythrocytes of patients with chronic hepatitis C (genotype 1b) before the initiation of antiviral therapy and after its completion. Methods. The study included 195 patients with chronic hepatitis C (genotype 1b). The levels of conjugated dienes, triene ketones, Schiff bases and oxidation index were measured after extraction of erythrocyte membranes with chloroform/isopropanol (2:1) mixture according to I.D. Stal’naya method modified by I.A. Volchegorskiy. All patients were allocated to four groups depending on the alanine transaminase activity prior to antiviral therapy: the first group (n=52) - with normal levels of the enzyme, the second (n=74) - with elevation not exceeding 3 normal ranges, the third group (n=42) - 3 to 5 normal ranges, the fourth group (n=27) - exceeding 5 normal ranges. Antiviral treatment included pegylated interferon alfa-2b and ribavirin for 48 weeks. Results. Level of conjugated dienes, triene ketones, Schiff bases and oxidation index were ranged from 0.71±0.04 to 1.02±0.03, from 0.45±0.08 to 0.94±0.02, 0.17±0.04 to 0.41±0.04 and from 0.91±0.13 to 1.91±0.03 units respectively, which was significantly higher than normal ranges and was directly dependent on the alanine transaminase activity. In addition, in patients who did not respond to antiviral treatment, these parameters were significantly higher compared to patients with immediate virologic response (p 0.01). Conclusion. Patients with chronic hepatitis C had increased levels of lipid peroxidation products in erythrocytes; their high content in patients who did not respond to antiviral treatment may be a predictive factor for antiviral treatment effectiveness.

The effect of volatile antioxidants of plant origin on leukemogenesis in mice

The effect of savory essential oil added with drinking water (150 ng/ml) or with feed (2.5 microg/g) on the lifetime of AKR mice and the parameters of oxidative stress in animal blood were investigated. It was found for the first time that long-term administration of an essential oil in low doses increased the average lifetime of mice by 20-35% and was accompanied by a decrease in the hemolysis level and the content of lipid peroxidation products in erythrocytes of mice, as well as alteration in the structural state of their membranes and stabilization of polyunsaturated fatty acids level in mice liver cells.

Lipid peroxidation and antioxidant status in patients with rheumatoid arthritis.

The changes in the erythrocyte lipid peroxidation products (MDA), levels of glutathione (GSH), ascorbic acid and plasma vitamin E (non enzymatic antioxidant parameters) and activities of antioxidant enzymes superoxide dismutase (SOD), glutathione peroxidase (GP(X)), catalase in erythrocytes and plasma glutathione - S - transferase (GST) activity were estimated in patients with rheumatoid arthritis. This work was undertaken to assess oxidative stress and anti oxidant status in patients with rheumatoid arthritis. It was observed that there was a significant increase in erythrocyte MDA levels, activities of SOD, GP(X), plasma GST and a significant decrease in erythrocyte GSH, ascorbic acid, plasma vitamin E levels and catalse activity in patients with rheumatoid arthritis when compared to controls. The results of our study suggests higher oxygen free radical production, evidenced by increased MDA and decreased GSH, ascorbic acid, vitamin E and Catalase activity, support to the oxidative stress in rheumatoid arthritis. The increased activities of antioxidant enzymes may be a compensatory regulation in response to increased oxidative stress.

Status of lipid peroxidation, glutathione, ascorbic acid, vitamin E and antioxidant enzymes in patients with osteoarthritis

The exact pro-oxidant and antioxidant status in osteoarthritis patients is still not clear. To add a new insight to the question, changes in the erythrocyte lipid peroxidation products (MDA), levels of glutathione (GSH), ascorbic acid and plasma vitamin E (nonenzymatic antioxidant parameters); and activities of antioxidant enzymes superoxide dismutase (SOD), glutathione peroxidase (GPX), catalase in erythrocytes and plasma glutathione - S - transferase (GST) were measured in patients with osteoarthritis. This work was undertaken to assess oxidative stress and antioxidant status in patients with osteoarthritis. The study was conducted in 20 patients and compared to controls. Levels of erythrocyte MDA, GSH, ascorbic acid, plasma vitamin E; and activities of antioxidant enzymes were measured in patients with osteoarthritis. Erythrocyte GSH was measured by the method of Beutler et al. Ascorbic acid levels were measured by the method of Tietz. Plasma vitamin E levels were measured by the method of Baker et al. MDA was determined as the measure of thio barbituric acid reactive substances (TBARS). SOD activity in the hemolysate was measured by the method of Misra and Fridovich. Activity of catalase was measured by the method of Beers and Sizer. GPX activity was measured as described by Paglia and Valentine in erythrocytes and Plasma GST activity was measured as described by Warholm et al. These parameters were measured in 20 patients and compared to controls. Statistical analysis between group 1 (controls) and group 2 (patients) was performed by the student's t - test using the stat -view package. It was observed that there was a significant increase in erythrocyte MDA levels; SOD, GPX and plasma GST activities; and a significant decrease in erythrocyte GSH, ascorbic acid, plasma vitamin E levels and catalase activity in patients with osteoarthritis when compared to controls. The results of our study suggest higher oxygen-free radical production, evidenced by increased MDA and decreased GSH, ascorbic acid, vitamin E and catalase activity, support to the oxidative stress in osteoarthritis. The increased activities of antioxidant enzymes may be a compensatory regulation in response to increased oxidative stress.

Melatonin protects against lipid peroxidation and membrane rigidity in erythrocytes from patients undergoing cardiopulmonary bypass surgery.

The first aim of this study was to test whether there are changes in erythrocyte lipid peroxidation products and membrane fluidity during cardiac surgery involving cardiopulmonary bypass. Secondly, in vitro tests were performed to examine whether melatonin alters induced lipid peroxidation and reduced membrane fluidity in erythrocytes from these patients. Fifteen patients undergoing cardiac surgery involving cardiopulmonary bypass (CPB) were selected. Five blood samples were taken at different times during surgery for analysis of thiobarbituric acid reactive substances (TBARS) content and membrane fluidity of the erythrocytes. TBARS are an index of the level of lipid peroxidation. The results revealed an increase in TBARS levels and a parallel decrease in erythrocyte membrane fluidity (increased membrane rigidity) after the onset of CPB with respect to the reference sample. Likewise, in vitro induction of lipid peroxidation in the erythrocyte samples from CPB patients followed a similar pattern. The changes in TBARS levels and membrane fluidity were suppressed by pre-incubation of erythrocytes with melatonin prior to the induction (by 70 microm Fe2+ + 250 microm ascorbate) of lipid peroxidation in a concentration-dependent manner. The results constitute a persuasive argument for the use of melatonin for preventive and therapeutic purposes during CPB.

Mechanisms of anemia in SHP-1 protein tyrosine phosphatase-deficient “viable motheaten” mice

Viable motheaten mice (abbreviated gene symbol me(v)) are deficient in SHP-1, a critical negative regulator of signal transduction in hematopoietic cells. These mice exhibit severe immune dysfunction accompanied by hyperproliferation of myeloid cells, widespread inflammatory lesions, and regenerative anemia. The aim of this study was to investigate the mechanisms underlying anemia in me(v)/me(v) mice. Multiple hematologic parameters, osmotic fragility, and erythropoietin levels were measured to characterize the anemia in me(v)/me(v) mice. B-cell-deficient me(v)/me(v) Igh-6(null) mice were generated to assess the role of anti-erythrocyte antibodies. Coombs assays and flow cytometry were carried out for detection of anti-erythrocyte antibodies. Oxidant production by macrophages, glutathione levels, and lipid peroxidation products in erythrocytes were measured, as was the impact of oxidant on the ultrastructure of me(v)/me(v) erythrocytes. Erythroid maturation and erythrocyte plasma membrane integrity were assessed with flow cytometry by evaluating CD71 expression and annexin V labeling. The regenerative anemia of me(v)/me(v) mice was associated with erythrocyte changes that were independent of the presence of anti-erythrocyte antibodies. Erythrocytes from me(v)/me(v) mice had increased fragility and heightened susceptibility to oxidant damage. Macrophages from me(v)/me(v) mice demonstrated a higher basal level of oxidant production and enhanced production after stimulation. Oxidant damage in me(v)/me(v) erythrocytes was evidenced by a significant elevation of lipid peroxidation and diminished levels of glutathione. Our results support the hypothesis that as a consequence of severe inflammatory disease, me(v)/me(v) erythrocytes are subject to exceptionally high oxidative stress resulting in oxidation of phospholipids in the erythrocyte membrane with subsequent hemolysis.

Influence of age on activities of antioxidant enzymes and lipid peroxidation products in erythrocytes and neutrophils of down syndrome patients

Thirty-seven individuals with Down syndrome (DS) were divided into four age categories: (i) 1 to < 6 years, (ii) 6 to < 13 years, (iii) 13 to < 20 years, and (iv) over 20 years. Activities of antioxidant enzymes found in individual age categories were different, but the differences between age groups were not statistically significant. We confirmed significantly higher activities of Cu/Zn superoxide dismutase (SOD) and glutathione peroxidase (GPx) in blood cells of people with DS as compared to 35 controls, which consisted, for the first time, of siblings of children with DS. No significant differences were found in activities of catalase and glutathione reductase in DS vs. controls. A significant difference was observed in serum concentration of malondialdehyde (MDA) in DS vs. controls (8.39 ± 0.34 μmol/l vs. 7.34 ± 0.27 μmol/l; p = .021) and concentration of MDA in erythrocytes of individuals with DS between the third and fourth age group ( p = .05). In DS persons, an elevated ratio of SOD to catalase plus GPx with respect to the controls in all age categories was found, suggesting oxidative imbalance, potentially contributing to accelerated aging observed in these persons.

Possible protective effect of kolaviron on CCl4-induced erythrocyte damage in rats.

The possible protective effect of kolaviron on rat erythrocytes following simultaneous administration of kolaviron (100 mg/kg of body weight/day) with carbon tetrachloride CCl4 (1.195 g/kg of body weight/day) by separate intraperitoneal injections was investigated. Kolaviron, a biflavonoid fraction of the defatted alcoholic extract of Garcinia kola seed, inhibits the accumulation of lipid peroxidation products in erythrocytes. A significant reduction (p < 0.05) by about 34%, of lipid peroxidation products was observed in erythrocytes of rats treated simultaneously with CCl4 and kolaviron when compared to CCl4-treated rats. Similarly, the significant increase (p < 0.05) in membrane cholesterol observed in CCl4-treated rats was significantly decreased (p < 0.05) in rats treated simultaneously with CCl4 and kolaviron. Therefore, there was no significant difference (p < 0.05) in cholesterol phospholipid ratio (C/P) of rats treated simultaneously with CCl4 and kolaviron, and the controls. Thus, kolaviron normalizes the CCl4-induced change in erythrocyte membrane composition. In addition, kolaviron antagonizes the effect of CCl4 on the activity of the membrane bound enzyme, Ca2+-ATPase. These results suggest that kolaviron protects erythrocyte membranes from free radical attack, on both lipids and proteins.

Influence of trolox derivative and N-acetylcysteine on surface charge density of erythrocytes in methanol intoxicated rats

Methanol is oxidized in vivo to formaldehyde and then to formate and these processes are accompanied by free radicals generation. This paper reports the effect of antioxidants: trolox derivative (U-83836E) and N-acetylcysteine (NAC) on lipid peroxidation, surface charge density and hematological parameters of erythrocytes from rats intoxicated with methanol (3.0 g/kg body weight). Methanol administration caused increase in erythrocyte lipid peroxidation products and changes in surface charge density. Ingestion of methanol with U-83836E and NAC partially prevented these methanol-induced changes. This suggests that U83836E and NAC act as effective antioxidants and free radicals scavengers. They may have efficacy in protecting free radical damage to erythrocytes following methanol intoxication.