Role of phospholipase A2 in expression of the scavenger pathway in cultured aortic smooth muscle cells stimulated with phorbol 12-myristate 13-acetate.
We have demonstrated that cultured intimal smooth muscle cells (SMC) from thickened intima can metabolize acetylated low-density lipoprotein (LDL) by a scavenger pathway, but medial SMC from normal arteries cannot. In this study we investigated the expression mechanism of the scavenger pathway in medial SMC using a phorbol ester. Medial SMC were incubated with 10(-10)-10(-7) M phorbol 12-myristate 13-acetate (PMA) for 1-24 h and then their degradation of 125I-labelled acetylated LDL was assayed. Unstimulated SMC degraded little acetylated LDL, but incubation for 24 h with PMA dose-dependently stimulated its degradation by SMC, the optimal PMA concentration being 1 x 10(-8) M. Induction of expression of the scavenger pathway required more than 4 h of incubation with PMA and was completely inhibited by cycloheximide. In addition expression of the scavenger pathway was not transient but stable. Induction of expression of the scavenger pathway by PMA was not inhibited by protein kinase C inhibitors, but was inhibited about 50% by phospholipase A2 inhibitors. The study, using various phorbol esters, indicated that induction of the scavenger pathway was well correlated with their ability to stimulate phospholipase A2 in medial SMC but not with their ability to activate protein kinase C. Moreover, incubation with exogenous phospholipase A2 (0.1-10 units/ml) or its product, lysophosphatidylcholine (0.01-100 micrograms/ml) dose-dependently increased degradation of 125I-labelled acetylated LDL in medial SMC. Lysophosphatidylcholine was most effective in various lysophospholipids. These results suggest that PMA induced the scavenger pathway in part by stimulating phospholipase A2 in medial SMC, and that a product, lysophosphatidylcholine, is a mediator of expression of the scavenger pathway.
Morisaki
N
N
Second Department of Internal Medicine, School of Medicine, Chiba University, Japan.
Yokote
K
K
Takahashi
K
K
Otabe
M
M
Saito
Y
Y
Yoshida
S
S
Ueda
S
S
D000818:Animals; D001013:Aorta, Thoracic / Q000187:drug effects / Q000378:metabolism*; D001692:Biological Transport; D002478:Cells, Cultured; D003513:Cycloheximide / Q000494:pharmacology; D016166:Free Radical Scavengers; D007457:Iodine Radioisotopes; D007700:Kinetics; D008077:Lipoproteins, LDL / Q000378:metabolism; D008244:Lysophosphatidylcholines / Q000494:pharmacology; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000378:metabolism*; D010703:Phorbol Esters / Q000494:pharmacology*; D010741:Phospholipases A / Q000096:biosynthesis*; D054467:Phospholipases A2; D011817:Rabbits; D013379:Substrate Specificity; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*
D016166:Free Radical Scavengers; D007457:Iodine Radioisotopes; D008077:Lipoproteins, LDL; D008244:Lysophosphatidylcholines; D010703:Phorbol Esters; C032629:acetyl-LDL; D003513:Cycloheximide; D010741:Phospholipases A; D054467:Phospholipases A2; D013755:Tetradecanoylphorbol Acetate