Abstract | INTRODUCTION:
Glutathione, a major cellular non- protein thiol (NPSH), serves a central role in repairing damage induced by cancer drugs, pollutants and radiation and in the detoxification of several cancer chemotherapeutic drugs and toxins. Current methods measure glutathione levels only, which require cellular extraction, rather than the glutathione recycling dependent antioxidant activity in intact cells. Here, we present a novel method using a bioactive probe of the oxidative pentose phosphate cycle, termed the OxPhos™ test, to quantify glutathione recycling dependent antioxidant activity in whole blood and intact human and rodent cells without the need for the isolation and cytoplasm extraction of cells. METHODS: OxPhos™ test kit (Rockland Immunochemicals, USA), which uses hydroxyethyldisulfide (HEDS) as a probe for the oxidative pentose phosphate cycle, was used in these studies. The results with OxPhos™ test kit in human blood and intact cells were compared with total thiol and high pressure liquid chromatography/electrochemical detection of HEDS metabolism. RESULTS: The OxPhos™ test measured glutathione-dependent antioxidant activity both in intact human and rodent cells and breast cancer patient's blood with a better correlation coefficient and biological variability than the thiol assay. Additionally, human blood and mammalian cells treated with various arsenicals showed a concentration-dependent decrease in activity. DISCUSSION:
|
Authors | Jie Li, Donglan Zhang, Pearl A Jefferson, Kathleen M Ward, Iraimoudi S Ayene |
Journal | Journal of pharmacological and toxicological methods
(J Pharmacol Toxicol Methods)
2014 Jan-Feb
Vol. 69
Issue 1
Pg. 39-48
ISSN: 1873-488X [Electronic] United States |
PMID | 24149024
(Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
|
Copyright | © 2013. |
Chemical References |
- Antioxidants
- Arsenicals
- Sulfhydryl Compounds
- Glutathione
|
Topics |
- Animals
- Antioxidants
(metabolism)
- Arsenicals
(pharmacology)
- Breast Neoplasms
(drug therapy, metabolism)
- CHO Cells
- Cell Line
- Cell Line, Tumor
- Cricetulus
- Female
- Glutathione
(metabolism)
- Humans
- Oxidation-Reduction
(drug effects)
- Pentose Phosphate Pathway
(drug effects)
- Sulfhydryl Compounds
(pharmacology)
|