linoleic-acid and phenanthrene

Smoking is a major risk factor for endothelial cell injury and subsequent coronary artery disease. Epidemiological studies implicate the phospholipase A2/arachidonic acid cascade in the mechanism by which smoking causes heart disease. However, specific components of cigarette smoke that activate this pathway have not been identified. The purpose of this study was to investigate the effects of polycyclic aromatic hydrocarbons contained in cigarette smoke on phospholipase A2 (PLA2) activity and apoptosis of human coronary artery endothelial cells. 1-methylanthracene (1-MA), phenanthrene (PA), and benzo(a)pyrene (B(a)P) caused significant release of 3H-arachidonate from endothelial cells. 1-MA and PA, but not B(a)P, also caused significant release of 3H-linoleic acid. Release of fatty acids from membrane phospholipids preceded the onset of apoptosis. 3H-arachidonate release and apoptosis induced by 1-MA, B(a)P, and PA were inhibited by methylarachidonoyl-fluorophosphonate, an inhibitor of Groups IV and VI PLA2s. Bromoenol lactone, an inhibitor of Group VI enzymes, inhibited both 3H-arachidonate release and apoptosis induced by 1-MA and PA, but not B(a)P. MJ33, an inhibitor of the acidic calcium-independent PLA2, attenuated 3H-arachidonate release and apoptosis by PA, but not 1-MA or B(a)P. The presence of Groups IV and VI and the acidic iPLA2 in endothelial cells was demonstrated by reverse transcriptase-polymerase chain reaction and Western analysis. These data suggest that 1-MA, B(a)P and PA induce apoptosis of endothelial cells by a mechanism that involves activation of these three distinct isoforms of PLA2.

Topics: Anthracenes; Apoptosis; Arachidonic Acid; Benzo(a)pyrene; Cells, Cultured; Coronary Vessels; Endothelium, Vascular; Enzyme Inhibitors; Humans; Linoleic Acid; Models, Biological; Phenanthrenes; Phospholipases A; Phospholipases A2; Polycyclic Aromatic Hydrocarbons; Smoking

Phenanthrene, a polycyclic aromatic hydrocarbon, was efficiently oxidized by laccase in the presence of both 1-hydroxybenzotriazole and unsaturated lipids. 73% of initially added phenanthrene was degraded within 182 hours to give phenanthrene-9,10-quinone and 2,2'-diphenic acid as the major products. The system was also able to peroxidize linoleic acid to its corresponding hydroperoxides suggesting the involvement of lipid peroxidation in laccase catalyzed phenanthrene oxidation. Lipid peroxidation by laccase required 1-hydroxybenzotriazole and did not depend on Mn2+ and H2O2 suggesting that the chemical reactions involved differ from those previously reported for manganese peroxidase.

Topics: Basidiomycota; Biphenyl Compounds; Fatty Acids, Unsaturated; Fungal Proteins; Laccase; Linoleic Acid; Lipid Peroxidation; Oxidation-Reduction; Oxidoreductases; Phenanthrenes; Triazoles