Oxidative stress and the mobilisation of arachidonic acid in stimulated human platelets: role of hydroxyl radical.
Platelet functions, including eicosanoid biosynthesis, can be significantly altered by exposure to reactive oxygen species. We utilised the redox properties of the phenazine derivative, pyocyanin, to generate low micromolar levels of reactive oxygen species in order to investigate the metabolism of arachidonic acid by human platelets under oxidative stress. Eicosanoid production by platelets, pre-labelled with [3H]arachidonic acid (AA) and stimulated with the calcium ionophore A23187, was inhibited in the presence of pyocyanin. In contrast, platelets pre-treated with pyocyanin and concurrently exposed to A23187 and AA showed no evidence of inhibition. Analysis of the free label content of labelled, pyocyanin-treated platelets after stimulation revealed diminished levels of total free label and a corresponding increase in labelled phospholipid. Prior treatment with the antioxidants, superoxide dismutase, catalase or the hydroxyl radical scavenger, mannitol, before the addition of pyocyanin afforded protection against loss of eicosanoid production and restored AA release. We conclude that hydroxyl radicals inhibit one or more steps in the cascade leading to phospholipase A2 activation and release of arachidonic acid from platelet phospholipid stores.
Muller
M
M
Centre for Infectious Diseases and Microbiology, University of Sydney, Westmead Hospital, NSW, Australia.
Sorrell
T C
TC
D000975:Antioxidants / Q000494:pharmacology; D016718:Arachidonic Acid / Q000378:metabolism*; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism*; D015777:Eicosanoids / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000737:chemistry / Q000378:metabolism; D017665:Hydroxyl Radical / Q000378:metabolism*; D015227:Lipid Peroxidation; D008353:Mannitol / Q000494:pharmacology; D009249:NADP / Q000378:metabolism; D018384:Oxidative Stress; D010743:Phospholipids / Q000378:metabolism; D011710:Pyocyanine / Q000378:metabolism / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism
D000975:Antioxidants; D015777:Eicosanoids; D010743:Phospholipids; D017382:Reactive Oxygen Species; D016718:Arachidonic Acid; D017665:Hydroxyl Radical; D008353:Mannitol; D009249:NADP; D011710:Pyocyanine; D006861:Hydrogen Peroxide