Journal article
Nitric Oxide-Induced Cytotoxicity: Involvement of Cellular Resistance to Oxidative Stress and the Role of Glutathione in Protection
Pediatric research, Vol.37(1), pp.41-49
01/1995
DOI: 10.1203/00006450-199501000-00010
PMID: 7700733
Abstract
ABSTRACT: A series of experiments were designed to examine the potential cytotoxicity of nitric oxide (NO), or reactive species derived from NO, in HA1 fibroblasts and H2O2-resistant variants of this cell line, designated OC14 cells. A 1-h exposure at 37°C to a 1.7 mM bolus dose of NO, prepared in N2-gassed medium, significantly reduced clonogenic survival in the HA1 fibroblasts line to 60% of control cells treated with N2-gassed medium alone. The OC14 cells were found to be completely resistant (100% survival) to NO-mediated injury in comparable experiments. A second set of experiments was designed to determine the role of the intracellular antioxidant, glutathione, in protection against NO-mediated injury. Depletion of total glutathione resulted in a significant reduction in HA1 and OC14 clonogenic survival to 8% and 50% when compared with respective control cells. The effect of total glutathione depletion on NO-initiated toxicity in HA1 cells was dose- and cell-density dependent and was observed to occur within 5 min of exposure to NO. Further evidence of cytotoxicity was demonstrated by loss of trypan blue dye exclusion properties in glutathione-depleted HA1 cells after NO exposure. Other experiments demonstrated that nitrate and nitrite exposure produced no cytotoxicity in glutathione-depleted HA1 cells and that coincubation of NO-saturated medium with oxyhemoglobin inhibited NO-induced cytotoxicity in glutathione-depleted HA1 cells. These results demonstrate that 1) nitric oxide, or an NO-derived reactive nitrogen species other than nitrites or nitrates, is responsible for reduction in clonogenic survival and trypan blue dye exclusion capabilities in vitro; 2) biochemical pathways associated with cellular resistance to oxidative stress also confer resistance to NO-mediated injury in this cell model; and 3) total glutathione content determines a significant portion of cell sensitivity to NO-mediated cytotoxicity.
Details
- Title: Subtitle
- Nitric Oxide-Induced Cytotoxicity: Involvement of Cellular Resistance to Oxidative Stress and the Role of Glutathione in Protection
- Creators
- M Whit Walker - University of VirginiaMichael T Kinter - University of VirginiaRobert J Roberts - University of VirginiaDouglas R Spitz - Washington University Medical Center
- Resource Type
- Journal article
- Publication Details
- Pediatric research, Vol.37(1), pp.41-49
- DOI
- 10.1203/00006450-199501000-00010
- PMID
- 7700733
- NLM abbreviation
- Pediatr Res
- ISSN
- 0031-3998
- eISSN
- 1530-0447
- Language
- English
- Date published
- 01/1995
- Academic Unit
- Pathology; Radiation Oncology; Fraternal Order of Eagles Diabetes Research Center
- Record Identifier
- 9984313095702771
Metrics
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