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Journal of Lipid Research, Vol 24, 1070-1076, Copyright © 1983 by Lipid Research, Inc.
DW Morel, JR Hessler and GM Chisolm
Low density lipoprotein (LDL) has been reported to be injurious or toxic to
cells in vitro. This injurious effect is, in some instances, due to
oxidation of the lipid moiety of the lipoprotein. The objectives of this
study were to determine if the oxidation rendering the lipoprotein toxic to
human skin fibroblasts occurred by free radical mechanisms, and if so,
which of the common free radical oxygen species were involved. The
selective free radical blockers or scavengers employed included superoxide
dismutase for superoxide, catalase for hydrogen peroxide, dimethylfuran for
singlet molecular oxygen, and mannitol for hydroxyl radical. The presence
during lipoprotein preparation of general free radical scavengers (vitamin
E, butylated hydroxytoluene) or the divalent cation chelator
ethylenediamine tetraacetic acid prevented the formation of cytotoxic low
density lipoprotein, while the simultaneous presence of superoxide
dismutase and catalase partially inhibited its formation. The results
indicate that superoxide and/or hydrogen peroxide are involved in the
formation of the toxic LDL lipid. The toxic action of oxidized LDL could
not be prevented by inclusion of antioxidants in the culture medium,
indicating that an oxidized lipid was responsible for cell injury rather
than free radicals generated in culture by the action of oxidized LDL.
Three separate assays for cell injury (enumeration of attached cells, cell
loss of lactate dehydrogenase into the culture medium, and trypan blue
uptake) indicated a sequence of events in which the fibroblasts are
injured, die, and then detach.
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Low density lipoprotein cytotoxicity induced by free radical peroxidation of lipid
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