Abstract
Using a chemiluminescent method, the consumption of HOCl/OCl was investigated during interaction with liposomes prepared from dimyristoylphos-phatidylcholine (DMPC) or egg yolk phosphatidylcholine (EYPC). The concentration of HOCl/OCl-decreased with time in the suspension of EYPC that contain unsaturated lipids and did not change in DMPC liposome suspensions. HOCl/OCl- was consumed more rapidly in peroxidized EYPC. The amount of double bonds was lowered by 40% in peroxidized liposomes and decreased by approximately one-third under the action of HOCl/OCl- in both native and peroxidized EYPC samples. Second-order rate constants for the interaction between HOCl and phospholipid double bonds of 0.50 M-1s-1 were calculated for native EYPC on basis of the consumption of HOCl/OCl- or from the decrease in concentration of double bonds. In peroxidized EYPC this reaction constant was similar as determined following changes in double bonds. It is concluded that the consumption of HOCl/OCl- increased in peroxidized liposomes due to additional reactions with lipid peroxidation products. tert-Butyl hydroperoxide and cumene hydroperoxide, or organic peroxides or epoxides (cis-9,10-epoxystearic acid; cholesterol-5α, bα-epoxide; trans-2,3-epoxy-butane; cis-2,3-epoxy-butane) were incorporated into liposomes and investigated in respect to their ability (1) to increase the consumption of HOCl/OCl- in DMPC liposomes, (2) to generate a non-enhanced chemiluminescence with HOCl/OCl- and (3) to evoke an accumulation of lipid peroxidation products (TBARS) in EYPC liposomes in the absence and presence of NaOCl. None of peroxides or epoxides tested showed any effect on the consumption of HOCI/OCl- or the generation of chemiluminescence. Nor increase of TBARS both in the absence or presence of HOCI/OCl-. In contrast, tert-butyl hydroperoxide and cumene hydroperoxide increased the consumption of HOCI/OCl- in DMPC liposomes and mediated a higher accumulation of TBARS in EYPC liposomes in the presence of HOCl/OCL- over the control. These data suggest that lipid peroxidation in EYPC can be initiated by the reaction of HOCI/OCL- with organic hydroperoxides.
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