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Detection of Oxidative Stress in Lymphocytes Using Dichlorodihydrofluorescein Diacetate

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Stress Response

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 99))

Abstract

Normal oxidative metabolism leads to the generation of reactive oxygen species (ROS), in particular, superoxide anion (O2−), and its dismutation product hydrogen peroxide (H2O2), which may escape from the electron-transport chain (1. In addition, oxidative stress may be generated through the action of specialized enzymes, e.g., NADPH oxidase, nitric oxide synthase, cycloxygenase, and lipoxygenase, which produce H2O2, O2−, NO, and lipid hydroperoxides [ROOH] (2,3). Toxicants can also induce oxidative stress by a variety of mechanisms. Compounds that uncouple or block the electron-transport chain lead to increased leakage of ROS from mitochondria to the cytosol. Redox cycling of metals can result in H2O2, hydroxyl radical (·OH), and thionyl radical (RS·) production, and can deplete cellular thiol pools (4). In addition, toxicants may directly inhibit antioxidant enzymes or compromise cellular reducing capacity by depletion of NAD(P)H, and glutathione (GSH) through the cytochrome P450 oxidoreductase and glutathione-S-transferase detoxification pathways, and through the action of the GSH peroxidase/reductase cycle (5). In cells with diminished natural antioxidant capacity, normal metabolic sources of ROS may overwhelm the redox balance and push cells into a condition of oxidative stress.

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Author information

Authors and Affiliations

  1. Department of Environmental Health, University of Washington, Seattle, WA

    Cecile M. Krejsa

  2. Immunology, Inflammation and Pulmonary Drug Discovery, Bristol-Myers Squibb Pharmaceutical Research Institute, Princeton, NJ

    Gary L. Schieven

Authors
  1. Cecile M. Krejsa
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  2. Gary L. Schieven
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Editor information

Editors and Affiliations

  1. ICRF Laboratories, University of Dundee, UK

    Stephen M. Keyse

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© 2000 Humana Press Inc., Totowa, NJ

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Krejsa, C.M., Schieven, G.L. (2000). Detection of Oxidative Stress in Lymphocytes Using Dichlorodihydrofluorescein Diacetate. In: Walker, J.M., Keyse, S.M. (eds) Stress Response. Methods in Molecular Biology™, vol 99. Humana Press. https://doi.org/10.1385/1-59259-054-3:35

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  • DOI: https://doi.org/10.1385/1-59259-054-3:35

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-611-6

  • Online ISBN: 978-1-59259-054-4

  • eBook Packages: Springer Protocols

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