Modulation of intracellular GSH in THP-1 cells during oxidative stress induced by AAPH

Type of content
Theses / Dissertations
Publisher's DOI/URI
Thesis discipline
Biochemistry
Degree name
Master of Science
Publisher
University of Canterbury. School of Biological Sciences
Journal Title
Journal ISSN
Volume Title
Language
Date
2006
Authors
Brown, Erin
Abstract

The human monocyte-derived THP-1 cell line was incubated with 10mM AAPH in Earle’s Balanced Salt Solution at 37°C for up to 24 hours. Protein hydroperoxide formation occurred after an 8 hour lag phase which corresponded to glutathione loss observed in the cells. SDS-Page analysis confirmed protein degradation occurred after 6 hours. Cell viability measured by the MTT reduction assay also dropped after 8 hours. Reduction of intracellular glutathione levels using BSO caused reduction of the lag phase seen in protein hydroperoxide formation. Cell viability of BSO-treated cells was lower than control cells, indicating the initiation of apoptotic events. Flow cytometry analysis showed no difference between BSO-treated and control cells, indicating that GSH levels do not have an effect on the type of cell death observed in AAPH-induced oxidative damage on THP-1 cells. These results confirmed previous data in the lab suggesting THP-1 cells undergo AAPH-induced necrosis as a result of cellular damage, including the loss of GSH and the formation of protein hydroperoxides.

Description
Citation
Keywords
monocyte-derived THP-1 cell, incubated, AAPH, modulation
Ngā upoko tukutuku/Māori subject headings
ANZSRC fields of research
Rights
Copyright Erin Brown