ascorbic-acid and 8-hydroxyguanosine

High-dose vitamin C therapy might mediate beneficial clinical effects by counteracting reactive oxygen species. However, concerns are raised whether this approach might provoke diametrical (ie pro-oxidative) effects. The objective was to determine ascorbyl free radical (AFR) concentrations and potential variables of pro-oxidative damage.. Crossover study; six healthy males received daily infusions of 750 or 7500 mg vitamin C for six consecutive days. Fasting concentrations of vitamin C and AFR were determined daily. On day 1, concentrations of vitamin C and AFR were measured at 0.25, 0.5, 1, 2, 4 and 8 h post infusion. Plasma concentrations of thiobarbituric acid-reactive substances (TBARS), tocopherol and urine concentrations of 8-oxoguanosine were determined on days 1 and 6.. Kinetic studies on day 1 showed that concentrations of vitamin C and AFR displayed parallel dose- and time-dependent kinetics and elimination was highly efficient. Vitamin C and AFR fasting concentrations on days 2-6 were slightly above the baseline, suggesting new, stable steady states. TBARS decreased in both groups, whereas tocopherol and 8-oxoguanosine concentrations remained unchanged.. Kinetics of AFR largely depend on plasma vitamin C concentrations and AFR is eliminated efficiently. Our data do not support induction of pro-oxidative effects in healthy volunteers given intravenous high-dose vitamin C.. Pascoe Pharmazeutische Präparate GmbH, Giessen, Germany.

Topics: Adult; Antioxidants; Ascorbic Acid; Cross-Over Studies; Dose-Response Relationship, Drug; Fasting; Free Radical Scavengers; Free Radicals; Guanosine; Humans; Infusions, Intravenous; Male; Oxidation-Reduction; Oxidative Stress; Prospective Studies; Reactive Oxygen Species; Thiobarbituric Acid Reactive Substances

This study was designed to examine the effect of 500 to 5,000 mg of ascorbic acid on DNA adducts, natural killer (NK) cell activity, programmed cell death, and cell cycle analysis of human peripheral blood leukocytes. According to our hypothesis, if ascorbic acid is a pro-oxidant, doses between 500 and 5,000 mg should enhance DNA adduct formation, decrease immune function, change the cell cycle progression, and increase the rate of apoptosis. Twenty healthy volunteers were divided into four groups and given either placebo or daily doses of 500, 1,000 or 5,000 mg of ascorbic acid for a period of 2 weeks. On days 0, 1, 7, 15, and 21, blood was drawn from them, and the leukocytes were separated and examined for intracellular levels of ascorbic acid, the level of 8-hydroxyguanosine, NK cell activity, cell cycle progression, and apoptosis. Depending on the subjects, between a 0% and a 40% increase in cellular absorption of ascorbic acid was observed when daily doses of 500 mg were used. At doses greater than 500 mg, this cellular absorption was not increased further, and all doses produced equivalent increases in ascorbic acid on days 1 to 15. This increase in cellular concentration of ascorbic acid resulted in no statistically meaningful changes in the level of 8-hydroxyguanosine, increased NK cytotoxic activity, a reduced percentage of cells undergoing apoptosis, and switched cell cycle phases from S and G2/M to G0/G1. After a period of 1 week, with no placebo or vitamin washout, ascorbic acid levels along with functional assays returned to the baseline and became equivalent to placebos. In comparison with baseline values, no change (not more than daily assays variation) was seen in ascorbate concentrations or other assays during oral placebo treatment. We concluded that ascorbic acid is an antioxidant and that doses up to 5,000 mg neither induce mutagenic lesions nor have negative effects on NK cell activity, apoptosis, or cell cycle.

Topics: Administration, Oral; Adult; Antioxidants; Apoptosis; Ascorbic Acid; Calcium Carbonate; Cell Cycle; Cytotoxicity, Immunologic; DNA Adducts; DNA Damage; Dose-Response Relationship, Drug; Female; Flow Cytometry; Guanosine; Humans; Intestinal Absorption; Intracellular Fluid; Kidney Function Tests; Killer Cells, Natural; Leukocytes; Liver Function Tests; Lymphocyte Count; Male; Middle Aged; Oxidation-Reduction; Urinalysis

There appears to be a paucity of data examining the effect of dietary antioxidants on levels of oxidative DNA damage in vivo, limiting evidence-based assessment of antioxidant efficacy, mechanisms and recommendation for optimal intake. We have examined levels of 8-oxo-2'-deoxyguanosine (8-oxodG) in mononuclear cell DNA, serum and urine from subjects undergoing supplementation with 500 mg/day vitamin C. Significant decreases in DNA levels of 8-oxodG were seen, correlating strongly with increases in plasma vitamin C concentration. Furthermore we established a timecourse for sequential, significant increases in serum and urinary 8-oxodG levels. These results illustrate, for the first time in humans, the kinetics of 8-oxodG removal and processing in vivo, suggesting a role for vitamin C in the regulation of DNA repair enzymes and thereby demonstrating a non-scavenging antioxidant effect.

Topics: 8-Hydroxy-2'-Deoxyguanosine; Adolescent; Adult; Antioxidants; Ascorbic Acid; Chromatography, High Pressure Liquid; Deoxyguanosine; Dietary Supplements; DNA Damage; DNA Repair; Enzyme-Linked Immunosorbent Assay; Female; Gas Chromatography-Mass Spectrometry; Guanosine; Humans; Leukocytes, Mononuclear; Male; Middle Aged; Oxidative Stress

Reactive oxygen species (ROS) have a negative impact on sperm DNA, leading to the formation of oxidative products such as 8-oxo-7,8-dihydroxyguanosine. This compound causes fragmentation and, thus, has a mutagenic effect. Patient treatment with oral antioxidant vitamins is, therefore, standard practice for male infertility, in an attempt to decrease formation of ROS and improve fertility. In this study, the DNA fragmentation index and the degree of sperm decondensation were measured using the sperm chromatin structure assay before and after 90 days treatment with antioxidant vitamins associated with zinc and selenium. Antioxidant treatment led to a decrease in sperm DNA fragmentation (-19.1%, P < 0.0004), suggesting that at least part of the decay was linked to ROS. However, it also led to an unexpected negative effect: an increase in sperm decondensation with the same order of magnitude (+22.8%, P < 0.0009). The opening of interchain disulphide bridges in protamines may explain this aspect, as antioxidant vitamins, especially vitamin C, are able to open the cystin net, thus interfering with paternal gene activity during preimplantation development. This observation might explain the discrepancy observed concerning the role of these antioxidant treatments in improving male fertility.

Topics: Adjuvants, Immunologic; Administration, Oral; Antioxidants; Ascorbic Acid; Disulfides; DNA Fragmentation; Fertilization in Vitro; Guanosine; Humans; Infertility, Male; Male; Oxidative Stress; Reactive Oxygen Species; Sperm Injections, Intracytoplasmic; Spermatozoa

We tested whether consecutive days of prolonged submaximal exercise would result in oxidant stress sufficient to alter blood antioxidant profiles, progressively change and exhaust blood and plasma antioxidants, and damage RNA. Eleven moderately trained males (24.3 +/- 1.1 yr) exercised 90 min at 65% peak O2 uptake on a cycle ergometer for 3 consecutive days. During day 1 exercise, blood reduced glutathione (GSH) declined 55 +/- 10% and oxidized glutathione (GSSG) increased 28 +/- 7% within 15 min. Total blood glutathione did not significantly change during exercise. GSH levels returned to baseline after 15 min of recovery. On day 3, preexercise GSH and GSSG levels were not significantly different from day 1 preexercise values; essentially similar results were obtained during exercise and recovery. During day 1 exercise, plasma total ascorbate (ascorbate + dehydroascorbate) increased from 53.8 +/- 9.3 to 59.0 +/- 11.3 microM, and percent reduced ascorbate increased from 77.6 +/- 9.3 to 87.3 +/- 9.7%. During day 3 exercise, plasma ascorbate changes were similar to those on day 1. Plasma vitamin E did not change due to exercise on either day 1 or 3. RNA adducts, urinary 8-hydroxyguanosine, did not change significantly due to exercise. Observed increases in GSH oxidation indicate the presence of oxidant stress during prolonged submaximal exercise. Similar redox changes on consecutive days of exercise, with recovery to preexercise values within 15 min, indicate no evidence of persistent or cumulative exercise effects on blood glutathione redox status.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adult; Antioxidants; Ascorbic Acid; Blood Glucose; Creatine Kinase; Exercise; Glutathione; Guanosine; Humans; L-Lactate Dehydrogenase; Lactates; Lactic Acid; Lipid Peroxides; Male; Oxygen Consumption; Pulmonary Gas Exchange; Reactive Oxygen Species; RNA; Stress, Physiological; Vitamin E

Recently we showed that ascorbate and 5-aminosalicylic acid (5-ASA) prevented 8-hydroxydeoxyguanosine (8-OHdG) formation in calf thymus DNA exposed to UV-visible light. However, the ultimate defense against oxidative DNA damage depends on an intracellular/intranuclear effect of the compounds. In the present study we investigated the effect of ascorbate and 5-ASA on 8-OHdG formation in V79 Chinese hamster cells exposed to light from a sun-lamp. Exposure for 1 min (4560 mJ/cm2) increased 8-OHdG formation in cellular DNA to 30-40 times background level. Preincubation of the cells with ascorbate or 5-ASA at concentrations of 0.1, 1 and 10 mM diminished the 8-OHdG formation to 0.67, 0.74 and 0.49 times controls (P < 0.05) for ascorbate respectively, and to 0.82, 0.66 and 0.33 times controls (P < 0.05), for 5-ASA. These findings demonstrate that both ascorbate and 5-ASA prevent oxidative DNA damage in cells by acting as intracellular/intranuclear antioxidants.

Topics: Aminosalicylic Acids; Animals; Ascorbic Acid; Cell Line; Cricetinae; Cricetulus; DNA; Guanosine; Light; Mesalamine; Ultraviolet Rays