Free radical-operated proteotoxic stress in macrophages primed with lipopolysaccharide.

Free Radical Biology & Medicine
Zili ZhaiDario C Ramirez

Abstract

The free-radical-operated mechanism of death of activated macrophages at sites of inflammation is unclear, but it is important to define it in order to find targets to prevent further tissue dysfunction. A well-defined model of macrophage activation at sites of inflammation is the treatment of RAW 264.7 cells with lipopolysaccharide (LPS), with the resulting production of reactive oxygen species (ROS). ROS and other free radicals can be trapped with the nitrone spin trap 5,5-dimethyl-1-pyrroline N-oxide (DMPO), a cell-permeable probe with antioxidant properties, which thus interferes with free-radical-operated oxidation processes. Here we have used immuno-spin trapping to investigate the role of free-radical-operated protein oxidation in LPS-induced cytotoxicity in macrophages. Treatment of RAW 264.7 cells with LPS resulted in increased ROS production, oxidation of proteins, cell morphological changes and cytotoxicity. DMPO was found to trap protein radicals to form protein-DMPO nitrone adducts, to reduce protein carbonyls, and to block LPS-induced cell death. N-Acetylcysteine (a source of reduced glutathione), diphenyleneiodonium (an inhibitor of NADPH oxidase), and 2,2'-dipyridyl (a chelator of Fe(2+)) prevented LPS-induced o...Continue Reading

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Citations

May 21, 2014·Inflammation Research : Official Journal of the European Histamine Research Society ... [et Al.]·Ruiming ChangKai Yu
Aug 28, 2013·Free Radical Biology & Medicine·Rheal A TownerHugo C Castro Faria-Neto
Dec 3, 2014·Free Radical Biology & Medicine·Yuya HorinouchiRonald P Mason
Mar 29, 2018·Inflammation Research : Official Journal of the European Histamine Research Society ... [et Al.]·M D MuñozD C Ramirez

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