Inhibition of oxidative insult in cultured cells by a novel 6-chromanol-containing antioxidant.
N18-RE-105 neuronal hybridoma cells were used in a cell culture system to evaluate the protective effects of a novel 6-chromanol-containing antioxidant, U78517F. First, the incorporation of the compound into the cells was evaluated, using a serum albumin carrier. Then the cells were exposed to peroxide-generating compounds, and the cell injury was estimated from the loss of alpha-aminoisobutyric acid (AIB) transport. We found that U78517F only protected the cells significantly when the degree of oxidative insult was below a certain limit; the measurable protection of cells by U78517F against either cumene hydroperoxide or H2O2 was limited to a narrow range of concentrations of the reactive oxygen species generator. Additionally, the protection provided by U78517F was largely localized to the cell membrane and did not extend to protection of mitochondrial function. The action of U78517 was fully consistent with a direct radical scavenging in the cells. The results indicate that the following factors must be taken into account for evaluation of antioxidants in cell culture: (a) the delivery of a compound to cells, especially when the compound is lipophilic; (b) the nature and extent of the oxidative insult used to evaluate protection; and (c) the location of the protective agent in the cells.
Decker
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DE
Upjohn Company, Upjohn Laboratories, Kalamazoo, MI 49001, USA.
Vroegop
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SM
Buxser
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SE
D026905:Amino Acid Transport Systems; D000621:Aminoisobutyric Acids / Q000378:metabolism; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D001555:Benzene Derivatives / Q000494:pharmacology; D002352:Carrier Proteins / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002839:Chromans / Q000737:chemistry / Q000494:pharmacology*; D016166:Free Radical Scavengers / Q000494:pharmacology*; D006825:Hybridomas; D006861:Hydrogen Peroxide / Q000494:pharmacology; D051379:Mice; D018384:Oxidative Stress / Q000187:drug effects*; D010879:Piperazines / Q000737:chemistry / Q000494:pharmacology*; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism
D026905:Amino Acid Transport Systems; D000621:Aminoisobutyric Acids; D000975:Antioxidants; D001555:Benzene Derivatives; D002352:Carrier Proteins; D002839:Chromans; D016166:Free Radical Scavengers; D010879:Piperazines; D017382:Reactive Oxygen Species; C066382:U 78517F; C100049:2-aminoisobutyric acid; C029141:2,2,5,7,8-pentamethyl-1-hydroxychroman; D006861:Hydrogen Peroxide; C007164:cumene hydroperoxide