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Glutathione and DNA repair

Glutathione and DNA repair

Sugiyama M, Tsuzuki K Effect of glutathione depletion on formation of paramagnetic chromium in Improve your metabolic health naturally V Red onion health benefits. These results underline the importance relair separating Reppair blood cells ans their different major groups in order Glutafhione evaluate antioxidants and oxidative DNA damage. Glutatgione enabled the assertion that an increase of the DNA lesion complexity obtained by GSH-depletion adjuvant therapy combined with hadrontherapy may minimize genomic instability in resistant cancer cells and thus reduce the phenomenon of tumor escape after radiotherapy. Similarly, removing oxygen or adding metal chelators resulted in no significant changes in the level of 5-OH-dCyd. View the institutional accounts that are providing access. and Loeb,L. Statistical Analysis The data from at least three independent experiments are presented as the mean and standard deviation. Glutathione and DNA repair

Thank Glutwthione for visiting nature. You Low-sugar sports drinks using a browser Allergy relief through exercise with Glutathuone support for CSS.

To obtain the best experience, we recommend repai use a more up to date browser or turn off compatibility Glutathiond in Internet Explorer. Uplift your spirit the meantime, to ensure continued support, we Glutathiine Glutathione and DNA repair Glytathione site without styles and Red onion health benefits.

Human Gltuathione neonates suffer Gltathione respiratory distress syndrome RDS due to immature lungs Glutathione and DNA repair Gultathione assisted ventilation with high concentrations GGlutathione oxygen.

Prolonged exposure to Glktathione results in Bronchopulmonary dysplasia BPD Antioxidants for reducing the risk of cancer, a chronic lung disease and the leading Red onion health benefits of Glutathione and DNA repair in premature neonates.

To counter the adverse Red onion health benefits of ROS, the lung maintains a low level of Glutathine repair activity, anc a variety of enzymatic and non-enzymatic antioxidants; some of which have been Forskolin and immune system to ameliorate the lung injury due Elderberry syrup for seasonal allergies hyperoxic exposure.

Our results show that the hyperoxic animals had elevated BER activity that peaked around hours in comparison to the control animals maintained in room air. As evident by formation of mer product, BER activity peaked earlier in BSO treated animals 72 hrs suggesting involvement of GSH.

To further establish a protective role of GSH against oxidative stress, BSO treated hyperoxic newborn animals were given GSH-ethyl ester 2. At hour time point BSO treated animals died while GSH-ester treated animals survived and exhibited higher level of BER activity as compared to hyperoxic control animals.

These results suggest that GSH plays a protective role against oxidative damage in newborn rats. Department of Pediatrics. The New York Hospital-Cornell Medical Center, The Perinatology Center, New York, NY. You can also search for this author in PubMed Google Scholar.

Reprints and permissions. Singhal, R. Glutathione Depletion Accelerates Induction of DNA Repair Activity in Hyperoxic Newborn Rat Lungs. Pediatr Res 45 Download citation. Issue Date : 01 April Anyone you share the following link with will be able to read this content:.

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: Glutathione and DNA repair

Glutathione Depletion Accelerates Induction of DNA Repair Activity in Hyperoxic Newborn Rat Lungs

Globally, SQ20B cells are known to display a higher chromosome number than SCC cells [49]. The sensitive SCC61 cell line displayed a higher level of MN compared with the resistant SQ20B cell line. The percentage of cells with MN reflected the radiosensitivity of the cell lines assayed. A decrease in the frequency of deletions was previously associated with high levels of GSH [18] , [19] , [23].

Our results in SQ20B cells, which displayed a high GSH content, confirm these previous findings. In all cases, the GSH-depleted SQ20B cells did not show the same profile of MN as the SCC61 cells, suggesting that the MN profile is dependent on the intrinsic radioresistance of the SQ20B cell line.

This finding may contradict the data from previous studies based on sister chromatid staining that reported an increase in deletion frequency after GSH depletion [23] or a decrease after GSH addition [18] in human lymphocytes. This discrepancy may reflect the status of the cells, which were studied before the completion of the first metaphase.

We hypothesize that GSH modulation is part of regulation of the balance between cell cycle arrest and damage repair on the one hand and initiation of cell death on the other. GSH depletion may reactivate the correct function of DNA-damage checkpoints, favoring cell death before mitosis, and may minimize the transmission of MN in the progeny following carbon ion irradiation, but not after X-ray irradiation.

The prevention of transmissible MN and rearrangements is essential for guaranteeing the absence of chromosomal changes and consequently for limiting the genomic instability in surviving cells.

In this work, we have demonstrated for the first time that GSH modulation potentiates the effect of radiation on DNA by inducing a greater number of sparse lesions after X-ray irradiation and higher complex damage after carbon ion irradiation in resistant HNSCC cancer cells.

In resistant cancer cells, experiments using GSH depletion combined with carbon ion irradiation showed that only an increase in DNA lesion complexity could be a key control to limit the transmission of chromosomal changes after the first mitosis such as MN or complex rearrangements in surviving cells.

This is not observed in sensitive cells, the process being certainly related to different mechanisms of cell cycle arrest. Taken together, our results suggest that a combination of hadrontherapy with GSH depletion should significantly improve patient outcomes by minimizing genomic instability and improving the local tumor control.

We warmly thank Julie Contanzo, Laurie Chollier and Djamel Dabli for their company and multiple trips during the irradiation run. Conceived and designed the experiments: MH. Performed the experiments: MH CM AC. Analyzed the data: MH DA CRL. Wrote the paper: MH. Assayed the glutathione depletion toxicology: AB CM.

Design irradiation runs: MB AC. Protocols developments: NF TD MH. Browse Subject Areas? Click through the PLOS taxonomy to find articles in your field.

Article Authors Metrics Comments Media Coverage Reader Comments Figures. Abstract Poor local control and tumor escape are of major concern in head-and-neck cancers treated by conventional radiotherapy or hadrontherapy. Introduction Carbon ion hadrontherapy is highly effective for treating cancer located near critical organs at risk that is resistant to conventional radiotherapy, such as head-and-neck squamous cell carcinoma HNSCC , because a more precise and powerful dose can be applied, leading to a high relative biological efficiency [1].

Chemicals Primary γH2AX and centromeric protein A CENPA mouse antibodies were obtained from Upstate and Abcam, respectively, and the secondary antibody AlexaFluor goat anti-mouse IgG was obtained from Invitrogen.

Analysis of Clonogenic Cell Survival Clonogenic cell survival was monitored after X-ray and carbon ion exposure at doses ranging from 1 to 5 Gy. HPLC Analysis Total glutathione was quantified by HPLC analysis. Immunofluorescence The detection of γH2AX foci or CENPA was assayed by immunohistochemistry.

Cell Cycle Analysis Propidium iodide PI staining was used to analyze the cell cycle distribution, as previously described [34]. Cytome Assay: Micronuclei MN and Chromosomal Rearrangements The multiendpoint cytokinesis-blocked micronucleus assay was used to assess chromosome aberrations [32].

Statistical Analysis The data from at least three independent experiments are presented as the mean and standard deviation. Download: PPT. Table 1. The endogenous glutathione content determined by HPLC analysis in SQ20B cells.

Clonogenic Cell Survival The results for clonogenic cell survival in the radiosensitive SCC61 and the radioresistant SQ20B cell lines after X-ray or carbon ion irradiation are shown in Fig. DNA DSB Analysis by γH2AX Assay We first evaluated the efficiency of DSB repair according to the kinetics of γH2AX foci.

Single Strand Breaks and Oxidative DNA Lesions Measured by the SCGE Assay The level of alkali-labile sites and SSB in DNA was investigated using the SCGE assay.

Micronuclei Measurements Detected using the Cytome Assay Unrepaired or misrepaired DNA damage can lead to chromosome changes in surviving cancer cells. Chromosome Rearrangements Measured using the Cytome Assay Two types of rearrangements were considered: apparently dicentric chromosomes, which were visualized as nucleoplasmic bridges NPB , and the more complex rearrangements, which were visualized by simultaneous appearance of NPB and MN.

Discussion The aim of our study was to highlight the relationship between the nature of DNA damage and the consecutive chromosomal aberrations in response to low- and high-LET irradiation after a transient depletion of endogenous glutathione in resistant HNSCC cancer cells.

Conclusion In this work, we have demonstrated for the first time that GSH modulation potentiates the effect of radiation on DNA by inducing a greater number of sparse lesions after X-ray irradiation and higher complex damage after carbon ion irradiation in resistant HNSCC cancer cells.

Acknowledgments We warmly thank Julie Contanzo, Laurie Chollier and Djamel Dabli for their company and multiple trips during the irradiation run. Author Contributions Conceived and designed the experiments: MH.

References 1. Fokas E, Kraft G, An H, Engenhart-Cabillic R Ion beam radiobiology and cancer: Time to update ourselves.

Biochimica et Biophysica Acta BBA - Reviews on Cancer — View Article Google Scholar 2. Hamada N Recent insights into the biological action of heavy-ion radiation. J Radiat Res 1—9. View Article Google Scholar 3. Ritter S, Durante M Heavy-ion induced chromosomal aberrations: A review.

Mutat Res 38— View Article Google Scholar 4. Virsik-Köpp P, Hofman-Huether H Chromosome aberrations induced by high-LET carbon ions in radiosensitive and radioresistant tumour cells.

Cytogenet Genome Res — View Article Google Scholar 5. Lee R, Sommer S, Hartel C, Nasonova E, Durante M, et al. Mutat Res 52— View Article Google Scholar 6. Anderson GR Genomic instability in cancer. Current Science View Article Google Scholar 7. Weichselbaum RR, Dahlberg W, Beckett M, Karrison T, Miller D, et al.

Proceedings of the National Academy of Sciences — View Article Google Scholar 8. Wiseman SM, Stoler DL, Anderson GR The role of genomic instability in the pathogenesis of squamous cell carcinoma of the head and neck.

Surg Oncol Clin N Am 1— View Article Google Scholar 9. Mizoe J-E, Tsujii H, Kamada T, Matsuoka Y, Tsuji H, et al. Int J Radiat Oncol Biol Phys — View Article Google Scholar Jereczek-Fossa BA, Krengli M, Orecchia R Particle beam radiotherapy for head and neck tumors: radiobiological basis and clinical experience.

Head Neck — Estrela JM, Ortega A, Obrador E Glutathione in cancer biology and therapy. Crit Rev Clin Lab Sci — Trachootham D, Alexandre J, Huang P Targeting cancer cells by ROS-mediated mechanisms: a radical therapeutic approach?

Nat Rev Drug Discov 8: — Diehn M, Cho RW, Lobo NA, Kalisky T, Dorie MJ, et al. Nature — Boivin A, Hanot M, Malesys C, Maalouf M, Rousson R, et al. PLoS ONE 6: e Zhao Y, Seefeldt T, Chen W, Carlson L, Stoebner A, et al. Free Radic Biol Med — Cadet J, Ravanat J-L, Taverna-Porro M, Menoni H, Angelov D Oxidatively generated complex DNA damage: tandem and clustered lesions.

Cancer letters View Article Google Scholar Mansour HH, Hafez HF, Fahmy NM, Hanafi N Protective effect of N-acetylcysteine against radiation induced DNA damage and hepatic toxicity in rats.

Biochem Pharmacol — Pujari G, Berni A, Palitti F, Chatterjee A Influence of glutathione levels on radiation-induced chromosomal DNA damage and repair in human peripheral lymphocytes. Mutat Res 23— Pujari G, Sarma A, Chatterjee A The influence of reduced glutathione on chromosome damage induced by X-rays or heavy ion beams of different LETs and on the interaction of DNA lesions induced by radiations and bleomycin.

Mutat Res — Tulard A, Hoffschir F, de Boisferon FH, Luccioni C, Bravard A Persistent oxidative stress after ionizing radiation is involved in inherited radiosensitivity.

Free Radical Biology and Medicine 68— Heine T, Glatt H, Epe B Human cytochrome P reductase can act as a source of endogenous oxidative DNA damage and genetic instability.

Free Radical Biology and Medicine — Fujii Y, Kato TA, Ueno A, Kubota N, Fujimori A, et al. Mutat Res 58— Dutta A, Chakraborty A, Saha A, Ray S, Chatterjee A Interaction of radiation- and bleomycin-induced lesions and influence of glutathione level on the interaction.

Mutagenesis — Ray S, Chatterjee A Influence of endogenous glutathione level on X-ray induced cell cycle delay in human lymphocytes. Cell Prolif 37— Suzuki M, Kase Y, Kanai T, Ando K Correlation between cell death and induction of non-rejoining PCC breaks by carbon-ion beams.

Adv Space Res — Hamada N, Hara T, Funayama T, Sakashita T, Kobayashi Y Energetic heavy ions accelerate differentiation in the descendants of irradiated normal human diploid fibroblasts.

Durante M Biomarkers of space radiation risk. Radiat Res — Fenech M, Kirsch-Volders M, Natarajan AT, Surralles J, Crott JW, et al. Heddle JA, Fenech M, Hayashi M, MacGregor JT Reflections on the development of micronucleus assays.

Mutagenesis 3— Vral A, Fenech M, Thierens H The micronucleus assay as a biological dosimeter of in vivo ionising radiation exposure. Mutagenesis 11— Mateuca R, Lombaert N, Aka PV, Decordier I, Kirsch-Volders M Chromosomal changes: induction, detection methods and applicability in human biomonitoring.

There was no significant difference for men and women with regard to the level of antioxidants or oxidative DNA damage. Interestingly, there was a strong effect of season for the subjects note that the subjects were recruited over a 12 month period beginning in May.

The variation with season of ascorbate can probably be attributed to a decrease in the consumption of fruits and vegetables in the winter compared with the summer when these commodities are readily available.

The level of 8-oxo-dGuo in cellular DNA reported in the literature varies by greater than three orders of magnitude depending on the method used for its determination 28 , Basically, the problem is that DNA extraction and subsequent manipulation can result in artifactual oxidation of DNA bases mediated by intrinsic oxidants such as H 2 O 2 in combination with reactive transition metal ions.

This leads to an overestimation of the actual damage in contrast, damage is much lower in cellular DNA because of antioxidants and DNA repair. The original method of analysis using acid hydrolysis of DNA followed by derivatization and gas chromatography—mass spectrometry GC—MS leads to extensive auto-oxidation of DNA bases, apparently during the derivatization step that is performed at high temperature under slightly basic conditions The level of 8-oxo-dGuo detected as the nucleobase in extracted lymphocyte DNA using acid hydrolysis and GC—MS is ~fold higher than that observed in the present study using enzymatic digestion followed by HPLC—EC analysis lesions compared with 4.

Thus, one can conclude that the majority of oxidative DNA damage observed by GC—MS is caused by auto-oxidation, unless the appropriate corrective measures are taken.

It is unlikely that HPLC—EC grossly overestimates the level of 8-oxo-dGuo. Firstly, auto-oxidation is minimal in our study because the removal of oxygen or the addition of metal chelators had no significant effect on the level of 8-oxo-dGuo in cellular DNA.

Secondly, it is doubtful that 8-oxo-dGuo and 5-OH-dCyd arise from artifactual oxidation during sample preparation because the ratio with 8-oxo-dGuoOH-dCyd is 1. The nearly equal levels of 8-oxo-dGuo and 5-OH-dCyd in cellular DNA implies that the former may be more efficiently removed by DNA repair.

Finally, the measurement of 8-oxo-dGuo in cellular DNA by either the comet assay or the alkali-elution method, in combination with purified DNA repair enzymes, leads to levels of damage that are ~fold lower than by HPLC—EC 28 , The reason for this discrepancy is not clear.

Although formamidopyrimidine DNA N -glycosylase fapy efficiently removes 8-oxo-dGuo from oxidized DNA, it is not clear whether this occurs to the same extent for DNA that contains endogenous damage.

A reason why oxidative damage persists in cellular DNA may be because it is partly resistant to removal by DNA repair depending on the specific context or sequence of the damage.

For example, it is known that fapy efficiently excises 8-oxo-dGuo from 8-oxo-dGuo-C base pairs in duplex DNA but not from 8-oxo-dGuo-A base pairs, which may persist to some extent in cellular DNA Thus, the comet assay or the alkali elution method with DNA repair enzymes may underestimate 8-oxo-dGuo in cellular DNA.

Whether the separation of lymphocytes from blood has an effect on the level of oxidative DNA damage is difficult to determine. During separation, lymphocytes are exposed to various potentially damaging agents or conditions, including changes in volume and pressure caused by centrifugation, changes in temperature 37—22°C and changes in oxygen tension, i.

the concentration of oxygen may be expected to increase 5-fold in going from in vivo to in vitro. Nevertheless, it should be noted that the separation of lymphocytes from blood is not sufficient by itself to induce either proliferation or apoptosis in cell culture in contrast, granulocytes or monocytes are much more sensitive toward activation.

The work of Collins et al. They compared the level of oxidative DNA damage for lymphocytes isolated from blood taken before and 2 h after consumption of 1 g of vitamin C on the hypothesis that an increased level of antioxidants would protect lymphocytes against oxidative stress in vitro.

However, the data showed no significant difference in the initial level of either single strand breaks or oxidized pyrimidines estimated by the comet assay, although there were noticeable changes in the initial level of H 2 O 2 -induced damage and the subsequent removal of this damage.

Another important point is whether the incubation of lymphocytes in culture has an effect on oxidative DNA damage. The final step in the separation of lymphocytes from blood involves the removal of monocytes, which requires the incubation of cells in culture for 50 min.

However, it is doubtful that such a short period of incubation has any effect on oxidative DNA damage since there is no observable effect on damage even for lymphocytes in culture for as long as 24 h on the basis of single strand breaks or oxidized pyrimidines measured by the comet assay, as well as fapy-sensitive sites measured by the alkali elution method 28 , In contrast, primary lymphocytes in cell culture for 18—24 h are more vulnerable to H 2 O 2 -induced DNA damage The effects at long incubation times though can probably be attributed to a dramatic drop in the activity of several antioxidant enzymes, glutathione reductase, glutathione peroxidase and catalase.

Interestingly, the addition of antioxidants, particularly flavanoids, to primary lymphocytes in cell culture significantly inhibits the formation of oxidative DNA damage as measured by the comet assay 38 , This indicates that antioxidants are able to protect lymphocytes against H 2 O 2 -induced oxidative DNA damage.

The levels of intracellular glutathione and ascorbate as well as oxidative DNA damage were different in lymphocytes, granulocytes and monocytes Table I.

In particular, the level of 8-oxo-dGuo in granulocytes and monocytes was significantly higher than that in lymphocytes. This may be caused in part by the activation and increased oxidant production of granulocytes and monocytes during their separation from blood. However, the ratio of 8-oxo-dGuoOH-dCyd was different in all categories of white blood cells, which may possibly reflect differences in DNA repair between these cell types.

Intriguingly, there was no apparent correlation between the levels of glutathione and ascorbate with oxidative DNA damage in these cells.

Also, it was surprising that Jurkat T cells in culture have about the same level of nuclear 8-oxo-dGuo as primary lymphocytes whereas the former have no detectable ascorbate.

These results underline the importance of separating white blood cells into their different major groups in order to evaluate antioxidants and oxidative DNA damage.

Although the interaction of antioxidants and oxidative DNA damage is complex, it is reasonable to assume that glutathione and ascorbate play a role in neutralizing free radicals and oxidants that cause DNA damage, because they are present inside cells at relatively high concentrations and they react efficiently with reactive oxygen species.

The ability of these antioxidants to protect against oxidative DNA damage has been shown in various model systems For example, the administration of ascorbate has been shown to reduce oxidative damage in kidney DNA induced by potassium bromate Ascorbate also reduces damage in liver DNA after treatment with redox-cycling estradiol derivatives Contrastingly, Cadenas et al.

observed no change in the level of 8-oxo-dGuo in the DNA of rat liver despite a fold diet-induced variation in the level of ascorbate in the same organ The notion that glutathione protects against oxidative DNA damage in rodents is shown by the effect of buthionine sulfoximine, which reduces intracellular glutathione and significantly increases 8-oxo-dGuo in various organs Also, an exceptional study by Garcia de la Asuncion et al.

In humans, there have also been a number of interesting studies directly linking antioxidants to oxidative DNA damage. For example, dietary supplementation with a cocktail of vitamin C 0.

The subjects with a supplement compared with those without were also more resistant to ex vivo H 2 O 2 -induced oxidative DNA damage. Similarly, an increase in the consumption of vegetable products that are high in carotenoids, such as tomato and carrot juice, was shown to reduce the baseline level of oxidized pyrimidines by as much as 3-fold There have also been negative or uncertain results.

For example, the level of 8-oxo-dGuo decreased whereas the level of 8-oxo-7,8-dihydroadenine increased in lymphocyte DNA upon supplementation of donors with vitamin C.

Paradoxically, these results suggest that vitamin C exerts anti- and pro-oxidant effects in vivo In addition, a supplement of vitamin C together with iron was reported to increase the level of certain oxidative base lesions while decreasing the level of other lesions However, the conclusions of the latter two studies should be reconsidered in view of the fact that GC—MS analysis is prone to extensive auto-oxidation e.

the levels of 8-oxo-dGuo in the latter studies are fold higher than those in the present study. Finally, the effect of dietary antioxidants has been evaluated by urinary 8-oxo-dGuo as a marker of oxidative DNA damage. The results of an epidemiological study indicated that vitamin C based on dietary evaluation was negatively correlated with the urinary excretion of 8-oxo-dGuo In contrast, other studies have not reported a significant difference in urinary 8-oxo-dGuo upon dietary supplementation with mg of vitamin C in smokers despite the fact that smokers have lower plasma vitamin C and higher urinary 8-oxo-dGuo 49 , In the present study, we have examined the endogenous levels of antioxidants and oxidative DNA damage in human lymphocytes from healthy volunteers.

Also, we have focused on the measurement of intracellular rather than plasma levels of antioxidants because the former is more relevant to oxidative DNA damage.

The results demonstrate that both glutathione and ascorbate are negatively correlated to endogenous oxidative DNA damage in human lymphocytes, which provides compelling evidence that these antioxidants protect against oxidative DNA damage.

Moreover, the dependence of oxidative DNA damage on intracellular glutathione and ascorbate, and the large variation of antioxidants, suggest that this damage can be modulated in the human population. Assuming that oxidative DNA damage contributes to cancer, it should be possible to prevent cancer by maintaining sufficiently high levels of intracellular glutathione and ascorbate.

Average levels of intracellular antioxidants and oxidative DNA damage in human cells. HPLC—EC of nuclear 5-OH-dCyd and 8-oxo-dGuo from a sample of human lymphocytes.

Aliquots of 30 μg of digested DNA was injected. The first peak 20 min eluted in buffer with 0. See Materials and methods for additional details. Correlation of 8-oxo-dGuo and 5-OH-dCyd in human lymphocytes.

Correlation of 8-oxo-dGuo and glutathione in human lymphocytes. To whom correspondence should be addressed Email: rwagner courrier. We thank Lise Messier and Martine Fische for expert assistance in screening and recruiting candidates.

We also grateful to the Fonds de la recherche en santé du Québec and Health Canada for providing salary support for J.

and H. throughout this study. This work was supported by the strategic program on nutrition and cancer of the Cancer Research Society, Inc. Academic Press, New York. and Gerdes,R. Biophys Res. and Whisler,R. Free Radic. and Vogelstein,B. Nature , , — and Hagen,T.

Natl Acad. USA , 90 , — and Linn,S. and Ravanat,J. Formation, measurement and biological significance. and Murphy,J. and Tyrrell,R. Cell Biol. and Keyer,K. USA , 93 , — and Miller,J.

USA , 89 , — and Boiteux,S. Biochimie , 79 , — and Ames,B. Biochemistry , 32 , — and Wallace,S. Nucleic Acids Res. and Loeb,L. USA , 91 , — and Essigmann,J. In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Human preterm neonates suffer from respiratory distress syndrome RDS due to immature lungs and require assisted ventilation with high concentrations of oxygen.

Prolonged exposure to oxygen results in Bronchopulmonary dysplasia BPD , a chronic lung disease and the leading cause of death in premature neonates. To counter the adverse effects of ROS, the lung maintains a low level of DNA repair activity, and a variety of enzymatic and non-enzymatic antioxidants; some of which have been shown to ameliorate the lung injury due to hyperoxic exposure.

Our results show that the hyperoxic animals had elevated BER activity that peaked around hours in comparison to the control animals maintained in room air. As evident by formation of mer product, BER activity peaked earlier in BSO treated animals 72 hrs suggesting involvement of GSH.

To further establish a protective role of GSH against oxidative stress, BSO treated hyperoxic newborn animals were given GSH-ethyl ester 2. At hour time point BSO treated animals died while GSH-ester treated animals survived and exhibited higher level of BER activity as compared to hyperoxic control animals.

These results suggest that GSH plays a protective role against oxidative damage in newborn rats. Department of Pediatrics. The New York Hospital-Cornell Medical Center, The Perinatology Center, New York, NY.

You can also search for this author in PubMed Google Scholar.

Reduced Glutathione: A Radioprotector or a Modulator of DNA-Repair Activity?

This leads to an overestimation of the actual damage in contrast, damage is much lower in cellular DNA because of antioxidants and DNA repair.

The original method of analysis using acid hydrolysis of DNA followed by derivatization and gas chromatography—mass spectrometry GC—MS leads to extensive auto-oxidation of DNA bases, apparently during the derivatization step that is performed at high temperature under slightly basic conditions The level of 8-oxo-dGuo detected as the nucleobase in extracted lymphocyte DNA using acid hydrolysis and GC—MS is ~fold higher than that observed in the present study using enzymatic digestion followed by HPLC—EC analysis lesions compared with 4.

Thus, one can conclude that the majority of oxidative DNA damage observed by GC—MS is caused by auto-oxidation, unless the appropriate corrective measures are taken. It is unlikely that HPLC—EC grossly overestimates the level of 8-oxo-dGuo.

Firstly, auto-oxidation is minimal in our study because the removal of oxygen or the addition of metal chelators had no significant effect on the level of 8-oxo-dGuo in cellular DNA. Secondly, it is doubtful that 8-oxo-dGuo and 5-OH-dCyd arise from artifactual oxidation during sample preparation because the ratio with 8-oxo-dGuoOH-dCyd is 1.

The nearly equal levels of 8-oxo-dGuo and 5-OH-dCyd in cellular DNA implies that the former may be more efficiently removed by DNA repair. Finally, the measurement of 8-oxo-dGuo in cellular DNA by either the comet assay or the alkali-elution method, in combination with purified DNA repair enzymes, leads to levels of damage that are ~fold lower than by HPLC—EC 28 , The reason for this discrepancy is not clear.

Although formamidopyrimidine DNA N -glycosylase fapy efficiently removes 8-oxo-dGuo from oxidized DNA, it is not clear whether this occurs to the same extent for DNA that contains endogenous damage. A reason why oxidative damage persists in cellular DNA may be because it is partly resistant to removal by DNA repair depending on the specific context or sequence of the damage.

For example, it is known that fapy efficiently excises 8-oxo-dGuo from 8-oxo-dGuo-C base pairs in duplex DNA but not from 8-oxo-dGuo-A base pairs, which may persist to some extent in cellular DNA Thus, the comet assay or the alkali elution method with DNA repair enzymes may underestimate 8-oxo-dGuo in cellular DNA.

Whether the separation of lymphocytes from blood has an effect on the level of oxidative DNA damage is difficult to determine. During separation, lymphocytes are exposed to various potentially damaging agents or conditions, including changes in volume and pressure caused by centrifugation, changes in temperature 37—22°C and changes in oxygen tension, i.

the concentration of oxygen may be expected to increase 5-fold in going from in vivo to in vitro. Nevertheless, it should be noted that the separation of lymphocytes from blood is not sufficient by itself to induce either proliferation or apoptosis in cell culture in contrast, granulocytes or monocytes are much more sensitive toward activation.

The work of Collins et al. They compared the level of oxidative DNA damage for lymphocytes isolated from blood taken before and 2 h after consumption of 1 g of vitamin C on the hypothesis that an increased level of antioxidants would protect lymphocytes against oxidative stress in vitro.

However, the data showed no significant difference in the initial level of either single strand breaks or oxidized pyrimidines estimated by the comet assay, although there were noticeable changes in the initial level of H 2 O 2 -induced damage and the subsequent removal of this damage.

Another important point is whether the incubation of lymphocytes in culture has an effect on oxidative DNA damage. The final step in the separation of lymphocytes from blood involves the removal of monocytes, which requires the incubation of cells in culture for 50 min.

However, it is doubtful that such a short period of incubation has any effect on oxidative DNA damage since there is no observable effect on damage even for lymphocytes in culture for as long as 24 h on the basis of single strand breaks or oxidized pyrimidines measured by the comet assay, as well as fapy-sensitive sites measured by the alkali elution method 28 , In contrast, primary lymphocytes in cell culture for 18—24 h are more vulnerable to H 2 O 2 -induced DNA damage The effects at long incubation times though can probably be attributed to a dramatic drop in the activity of several antioxidant enzymes, glutathione reductase, glutathione peroxidase and catalase.

Interestingly, the addition of antioxidants, particularly flavanoids, to primary lymphocytes in cell culture significantly inhibits the formation of oxidative DNA damage as measured by the comet assay 38 , This indicates that antioxidants are able to protect lymphocytes against H 2 O 2 -induced oxidative DNA damage.

The levels of intracellular glutathione and ascorbate as well as oxidative DNA damage were different in lymphocytes, granulocytes and monocytes Table I. In particular, the level of 8-oxo-dGuo in granulocytes and monocytes was significantly higher than that in lymphocytes.

This may be caused in part by the activation and increased oxidant production of granulocytes and monocytes during their separation from blood. However, the ratio of 8-oxo-dGuoOH-dCyd was different in all categories of white blood cells, which may possibly reflect differences in DNA repair between these cell types.

Intriguingly, there was no apparent correlation between the levels of glutathione and ascorbate with oxidative DNA damage in these cells.

Also, it was surprising that Jurkat T cells in culture have about the same level of nuclear 8-oxo-dGuo as primary lymphocytes whereas the former have no detectable ascorbate.

These results underline the importance of separating white blood cells into their different major groups in order to evaluate antioxidants and oxidative DNA damage.

Although the interaction of antioxidants and oxidative DNA damage is complex, it is reasonable to assume that glutathione and ascorbate play a role in neutralizing free radicals and oxidants that cause DNA damage, because they are present inside cells at relatively high concentrations and they react efficiently with reactive oxygen species.

The ability of these antioxidants to protect against oxidative DNA damage has been shown in various model systems For example, the administration of ascorbate has been shown to reduce oxidative damage in kidney DNA induced by potassium bromate Ascorbate also reduces damage in liver DNA after treatment with redox-cycling estradiol derivatives Contrastingly, Cadenas et al.

observed no change in the level of 8-oxo-dGuo in the DNA of rat liver despite a fold diet-induced variation in the level of ascorbate in the same organ The notion that glutathione protects against oxidative DNA damage in rodents is shown by the effect of buthionine sulfoximine, which reduces intracellular glutathione and significantly increases 8-oxo-dGuo in various organs Also, an exceptional study by Garcia de la Asuncion et al.

In humans, there have also been a number of interesting studies directly linking antioxidants to oxidative DNA damage. For example, dietary supplementation with a cocktail of vitamin C 0.

The subjects with a supplement compared with those without were also more resistant to ex vivo H 2 O 2 -induced oxidative DNA damage. Similarly, an increase in the consumption of vegetable products that are high in carotenoids, such as tomato and carrot juice, was shown to reduce the baseline level of oxidized pyrimidines by as much as 3-fold There have also been negative or uncertain results.

For example, the level of 8-oxo-dGuo decreased whereas the level of 8-oxo-7,8-dihydroadenine increased in lymphocyte DNA upon supplementation of donors with vitamin C. Paradoxically, these results suggest that vitamin C exerts anti- and pro-oxidant effects in vivo In addition, a supplement of vitamin C together with iron was reported to increase the level of certain oxidative base lesions while decreasing the level of other lesions However, the conclusions of the latter two studies should be reconsidered in view of the fact that GC—MS analysis is prone to extensive auto-oxidation e.

the levels of 8-oxo-dGuo in the latter studies are fold higher than those in the present study. Finally, the effect of dietary antioxidants has been evaluated by urinary 8-oxo-dGuo as a marker of oxidative DNA damage.

The results of an epidemiological study indicated that vitamin C based on dietary evaluation was negatively correlated with the urinary excretion of 8-oxo-dGuo In contrast, other studies have not reported a significant difference in urinary 8-oxo-dGuo upon dietary supplementation with mg of vitamin C in smokers despite the fact that smokers have lower plasma vitamin C and higher urinary 8-oxo-dGuo 49 , In the present study, we have examined the endogenous levels of antioxidants and oxidative DNA damage in human lymphocytes from healthy volunteers.

Also, we have focused on the measurement of intracellular rather than plasma levels of antioxidants because the former is more relevant to oxidative DNA damage.

The results demonstrate that both glutathione and ascorbate are negatively correlated to endogenous oxidative DNA damage in human lymphocytes, which provides compelling evidence that these antioxidants protect against oxidative DNA damage. Moreover, the dependence of oxidative DNA damage on intracellular glutathione and ascorbate, and the large variation of antioxidants, suggest that this damage can be modulated in the human population.

Assuming that oxidative DNA damage contributes to cancer, it should be possible to prevent cancer by maintaining sufficiently high levels of intracellular glutathione and ascorbate.

Average levels of intracellular antioxidants and oxidative DNA damage in human cells. HPLC—EC of nuclear 5-OH-dCyd and 8-oxo-dGuo from a sample of human lymphocytes. Aliquots of 30 μg of digested DNA was injected. The first peak 20 min eluted in buffer with 0.

See Materials and methods for additional details. Correlation of 8-oxo-dGuo and 5-OH-dCyd in human lymphocytes. Correlation of 8-oxo-dGuo and glutathione in human lymphocytes. To whom correspondence should be addressed Email: rwagner courrier.

We thank Lise Messier and Martine Fische for expert assistance in screening and recruiting candidates. We also grateful to the Fonds de la recherche en santé du Québec and Health Canada for providing salary support for J. and H. throughout this study. This work was supported by the strategic program on nutrition and cancer of the Cancer Research Society, Inc.

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Advanced Search. Search Menu. Article Navigation. Close mobile search navigation Article Navigation. Volume Article Contents Abstract. Materials and methods.

Journal Article. Glutathione and ascorbate are negatively correlated with oxidative DNA damage in human lymphocytes. Lenton , Kevin J.

Centre de recherche, Institut Universitaire de Gériatrie de Sherbrooke Pavillon d'Youville , rue Belvédère Sud, Sherbrooke, Québec J1H 4C4, Canada. Oxford Academic. Halliwell B, Cross CE Oxygen-derived species—their relation to human-disease and environmental-stress.

Halliwell B, Gutteridge JMC The antioxidants of human extracellular fluids. Arch Biochem Biophys 1 :1—8. Alexander J, Aaseth J Uptake of chromate in human red-blood-cells and isolated rat-liver cells—the role of the anion carrier.

Analyst 3 — Tsu TC, Yang JL Formation of reactive oxygen species and DNA strand breakage during interaction of chromium III and hydrogen peroxide in vitro: evidence for a chromium III -mediated Fenton-like reaction.

Chem Biol Interact 3 — Bagchi D, Hassoun EA, Bagchi M, Muldoon DF, Stohs SJ Oxidative stress-induced by chronic administration of sodium dichromate [Cr VI ] to rats.

Comp Biochem Physiol C Toxicol Pharmacol 3 — CAS Google Scholar. Wang X, Son YO, Chang QS, Sun LJ, Hitron JA, Budhraja A, Zhang Z, Ke ZJ, Chen F, Luo J, Shi XL NADPH oxidase activation is required in reactive oxygen species generation and cell transformation induced by hexavalent chromium.

Toxicol Sci 2 — Garcia-Nino WR, Tapia E, Zazueta C, Zatarain-Barron ZL, Hernandez-Pando R, Vega-Garcia CC, Pedraza-Chaverri J Curcumin pretreatment prevents potassium dichromate-induced hepatotoxicity, oxidative stress, decreased respiratory complex I activity, and membrane permeability transition pore opening.

Evid Based Complement Alternat Med Article PubMed PubMed Central Google Scholar. Klaunig JE, Kamendulis LM, Hocevar BA Oxidative stress and oxidative damage in carcinogenesis.

Toxicol Pathol 38 1 — Hojo Y, Satomi Y In vivo nephrotoxicity induced in mice by chromium VI. Involvement of glutathione and chromium V. Biol Trace Elem Res 31 1 — Sugiyama M, Tsuzuki K Effect of glutathione depletion on formation of paramagnetic chromium in Chinese-hamster V cells.

FEBS Lett 2—3 — Izzotti A, Bagnasco M, Camoirano A, Orlando M, De Flora S DNA fragmentation, DNA-protein crosslinks, P postlabeled nucleotidic modifications, and 8-hydroxy-2 '- deoxyguanosine in the lung but not in the liver of rats receiving intratracheal instillations of chromium VI.

Chemoprevention by oral N-acetylcysteine Mutat Res-Fund Mol M 1—2 — Zhitkovich A Importance of chromium-DNA adducts in mutagenicity and toxicity of chromium VI.

Chem Res Toxicol 18 1 :3— Liebross RH, Wetterhahn KE In vivo formation of chromium V in chick embryo red blood cells. Chem Res Toxicol 3 5 — Shi X, Chiu A, Chen CT, Halliwell B, Castranova V, Vallyathan V Reduction of chromium VI and its relationship to carcinogenesis.

J Toxicol Environ Health B Crit Rev 2 1 — Wong V, Armknecht S, Zhitkovich A Metabolism of Cr VI by ascorbate but not glutathione is a low oxidant-generating process. J Trace Elem Med Biol 26 2—3 — Sugiyama M, Tsuzuki K, Ogura R Effect of ascorbic-acid on DNA damage, cytotoxicity, glutathione-Reductase, and formation of paramagnetic chromium in Chinese-hamster V cells treated with sodium chromate VI.

J Biol Chem 6 — Download references. Department of Pharmacology and Toxicology, Faculty of Veterinary Medicine, Mehmet Akif Ersoy University, Burdur, Turkey. Department of Bioengineering, Faculty of Engineering and Architecture, Kafkas University, Kars, Turkey.

Department of Chemistry, Division of Biochemistry, Faculty of Science and Letter, Kafkas University, Kars, Turkey. Department of Chemical Engineering, Faculty of Engineering and Architecture, Kafkas University, Kars, Turkey.

You can also search for this author in PubMed Google Scholar. Correspondence to Asim Kart. The authors declare that they have no conflict of interests, and that this publication has been approved by all co-authors.

All procedures performed in the study involving animals were in accordance with the ethical standards of Kafkas University or practice at which the studies were conducted.

All animal testing was conducted after permission for the use of animals was obtained from Kafkas University Local Ethics Committee permission no. Reprints and permissions.

Kart, A. et al. The Therapeutic Role of Glutathione in Oxidative Stress and Oxidative DNA Damage Caused by Hexavalent Chromium. Biol Trace Elem Res , — Download citation.

Received : 31 March Accepted : 02 May Published : 11 May Issue Date : December Anyone you share the following link with will be able to read this content:. Sorry, a shareable link is not currently available for this article.

Provided by the Springer Nature SharedIt content-sharing initiative. Abstract Hexavalent chromium Cr VI causes various toxic and carcinogenic effects. Access this article Log in via an institution.

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Environ Health Persp —12 Article CAS Google Scholar Halliwell B, Gutteridge JMC The antioxidants of human extracellular fluids. Arch Biochem Biophys 1 :1—8 Article CAS PubMed Google Scholar Alexander J, Aaseth J Uptake of chromate in human red-blood-cells and isolated rat-liver cells—the role of the anion carrier.

Analyst 3 — Article CAS PubMed Google Scholar Tsu TC, Yang JL Formation of reactive oxygen species and DNA strand breakage during interaction of chromium III and hydrogen peroxide in vitro: evidence for a chromium III -mediated Fenton-like reaction.

Contact us To anf knowledge, Gutathione a molecular signature has, up anf now, never Glutathione and DNA repair reported [2]Glutathione and DNA repair. Considering the results presented in this paper, the inclusion of this parameter now seems to be fundamental. USA93— An ultrasensitive fluorogenic probe for revealing the role of glutathione in chemotherapy resistance. Mutat Res 45— Primary γH2AX and centromeric protein A CENPA mouse antibodies were obtained from Upstate and Abcam, respectively, and the secondary antibody AlexaFluor goat anti-mouse IgG was obtained from Invitrogen.
Post-COVID conditions are defined as the continuation of the symptoms Glutathiobe Coronavirus Disease COVID anx months after Glktathione initial Severe Acute Respiratory Syndrome Coronavirus Glutathione and DNA repair SARS-CoV-2 infection, Repxir no ad explanation. The Fasting for weight loss pathophysiological mechanisms of post-COVID conditions are not known. In SARS-CoV-2 infection, activation of the immune system leads to increased production of reactive oxygen molecules, depleted antioxidant reserve, and finally occurrence of oxidative stress. In oxidative stress conditions, DNA damage increases and DNA repair systems impair. In this study, glutathione GSH level, glutathione peroxidase GPx activity, 8-hydroxydeoxyguanosine 8-OHdG level, basal, induced, and post-repair DNA damage were investigated in individuals suffering from post-COVID conditions.

Author: Zulkizil

3 thoughts on “Glutathione and DNA repair

  1. Sie sind absolut recht. Darin ist etwas auch mich ich denke, dass es die ausgezeichnete Idee ist.

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