Inhibitory effect of docosahexaenoic acid-containing phospholipids on 5-lipoxygenase in rat basophilic leukemia cells.
5-Lipoxygenase has been recognized to be an important enzyme that catalyzes the first step in leukotriene production. In this study we examine whether or not phospholipids containing docosahexaenoic acid (DHA) affect 5-lipoxygenase activity of a rat basophilic leukemia cell line (RBL-1). Among the synthesized phospholipids examined, 1-oleoyl-2-docosahexaenoyl-sn-glycero-3-phosphocholine (1-oleoyl-2-DHA-PC) was found to be the most potent inhibitor of 5-lipoxygenase. The inhibition was dose-dependent and the ID50 value was 4.0 microM. When the fatty acid at the sn-2-position was replaced by other unsaturated fatty acids, the inhibitory activity decreased with decreasing numbers of both carbon atoms and double bonds in the fatty acids. Substitution at the 1-position of the DHA-containing PC also affected the inhibitory potency. If oleic acid was substituted with palmitic acid, the inhibition activity was completely abolished. Lineweaver-Burk plot analysis showed that the inhibition of 5-lipoxygenase by 1-oleoyl-2-DHA-PC was non-competitive. The inhibition by this synthesized phospholipid was very specific to 5-lipoxygenase; that is, it did not extend to fatty acid cyclooxygenase, 12-lipoxygenase or 15-lipoxygenase. These results suggest that endogenously existing DHA-containing phospholipids may affect 5-lipoxygenase activity and thus control leukotriene biosynthesis in vivo.
Matsumoto
K
K
Section of Physiological Chemistry, Faculty of Dentistry, Tokyo Medical and Dental University, Japan.
Morita
I
I
Hibino
H
H
Murota
S
S
D000818:Animals; D004281:Docosahexaenoic Acids / Q000494:pharmacology*; D015471:Leukemia, Basophilic, Acute / Q000201:enzymology*; D016859:Lipoxygenase Inhibitors / Q000494:pharmacology*; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors*; D010713:Phosphatidylcholines / Q000494:pharmacology; D010743:Phospholipids / Q000494:pharmacology*; D051381:Rats; D013329:Structure-Activity Relationship; D014407:Tumor Cells, Cultured
D016859:Lipoxygenase Inhibitors; D009363:Neoplasm Proteins; D010713:Phosphatidylcholines; D010743:Phospholipids; D004281:Docosahexaenoic Acids; C084316:1-oleoyl-2-docosahexaenoyl phosphatidylcholine