Relative susceptibility of lipids to peroxidation in intact erythrocytes

Factors relating the site and metabolic state of erythrocyte lipids to their susceptibility to oxidative stress are poorly understood. Therefore red cell lipids were labeled with serum albumin-bound (carbon 14 [ul]) fatty acids in timed experiments. Susceptibility of sequentially labeled lipids to peroxidation in intact cells was determined by the percent conversion of the [14C]lipid to [14C]malondialdehyde (MDA). Other effects of alteration of incubation times could be ruled out by the equality of the conversion of total cell lipid to MDA. Polyunsaturated fatty acids but not other fatty acids served as MDA precursors. In 12 normal subjects the percentage of [14C]arachidonate converted to MDA after 15 minutes of labeling (3.08% +/- 0.69%) exceeded the percentage after 3 hours of labeling (2.01% +/- 0.83%) (paired, two-sided t test, p less than 0.0001). This susceptibility was caused by presence of rapidly labeled polyunsaturated fatty acids not extractable with defatted albumin (F2). Depletion of erythrocyte adenosine triphosphate before incubation favored accumulation of F2 and increased percentage conversion to MDA. Peroxidative susceptibility is determined by the metabolic state of the cell and the site of the peroxidized lipid within the intracellular pools.