Lipid-derived Radical Formation Research Articles

Free-Radical-Mediated Formation Mechanism of Polar Polymeric Triglycerides in Vegetable Oil Studied by Electron Spin Resonance and High-Performance Liquid Chromatography.

The free-radical-mediated formation mechanism of polar polymeric triglycerides (TAGs) was derived based on the formation of lipid-derived radicals and the degradation of TAGs in palm oil (PO), rapeseed oil (RO), and sunflower oil (SO). The experimental spectra were simulated by alkoxyl, alkyl, and 5-dimethyl-1-pyrroline N-oxide (DMPO)-oxidized adducts. DMPO-oxidized adducts were the main radical adducts in the initial stage. Then, alkyl radical adducts became the dominating radical adducts after 12 min in PO and RO. The intensity of alkyl radical adducts was the highest in SO. Therefore, based on the bimolecular reaction, polar polymeric TAGs were mainly bonded by -C-O-O-C- in the initial stage and then by -C-C- and -C-O-C- after 30 min. Besides, according to the correlation analysis between the amounts of polar polymeric TAGs and the degradation of TAGs, the main structures of polar polymeric TAGs in PO, RO, and SO were POL-LOP, POL-OOP, and POO-OOP; OLL-LnLO, OLLn-OLnO, OOO-OLO, and OLLn-OOO; and LLL-LLO, LLL-LLL, and OLL-LLO, respectively.

Acidic pH amplifies iron-mediated lipid peroxidation in cells

The goal of our study was to investigate the mechanism by which changes in extracellular pH influence lipid peroxidation processes. Ferrous iron can react with hydroperoxides, via a Fenton-type reaction, to initiate free radical chain processes. Iron is more soluble at lower pH values, therefore we hypothesized that decreasing the environmental pH would lead to increased iron-mediated lipid peroxidation. We used Photofrin, a photosensitizer that produces singlet oxygen, to introduce lipid hydroperoxides into leukemia cells (HL-60, K-562, and L1210). Singlet oxygen reacts with the PUFA of cells producing lipid hydroperoxides. Using EPR spin trapping with POBN, free radical formation from HL-60 cells was only detected when Photofrin, light, and ferrous iron were present. Free radical formation increased with increasing iron concentration; in the absence of extracellular iron, radical formation was below the limit of detection and lipid hydroperoxides accumulated in the membrane. In the presence of iron, lipid-derived radical formation in cells is pH dependent; the lower the extracellular pH (7.5–5.5), the higher the free radical flux; the lower the pH, the greater the membrane permeability induced in K-562 cells, as determined by trypan blue dye exclusion. These data demonstrate that lipid peroxidation processes, mediated by iron, are enhanced with decreasing extracellular pH. Thus, acidic pH not only releases iron from “safe” sites, but this iron will also be more damaging.

Singlet Oxygen Toxicity Is Cell Line‐dependent: A Study of Lipid Peroxidation in Nine Leukemia Cell Lines

Singlet oxygen (1O2) can be quenched by water, lipids, proteins, nucleic acids and other small molecules. Polyunsaturated fatty acids (PUFA) of cells principally quench 1O2 by chemical mechanisms, producing lipid hydroperoxides, while proteins physically and chemically quench 1O2. Because cell lines can have different PUFA and protein levels, we hypothesized that 1O2 toxicity will vary between cell lines. We used Photofrin as a source of 1O2. Exposure of nine different leukemia cell lines (CEM, HEL, HL-60, K-562, KG-1, L1210, Molt-4, THP-1 and U-937) to Photofrin and light results in changes in membrane permeability (trypan blue) that vary with cell line. The greater the lipid content of the cell line, the less susceptible they are to membrane damage. When the cell media was supplemented with docosahexaenoic acid (DHA, 22:6), the overall unsaturation of cellular lipids increased. Photofrin and light resulted in increased radical formation in these supplemented cells compared to controls; however, there was no difference in membrane permeability between DHA-supplemented and control cells. Lipid-derived radical formation (electron paramagnetic resonance spin trapping) was cell line dependent; but no correlation between lipid content of cells and radical formation was found. However, we found that the greater the protein content of cells the more they were protected against membrane damage induced by Photofrin photosensitization. This suggests that cellular proteins are a key target for 1O2-mediated toxicity. A remarkable observation is that cell size correlates inversely with ability of cells to cope with a given flux of 1O2.