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J. Biol. Chem., Vol. 266, Issue 15, 9419-9427, 05, 1991
CF Reilly and RC McFall
Bovine vascular smooth muscle cells (SMC) were examined for production of
plasminogen activator inhibitor-1 (PAI-1) which may play a key role in
regulating the fibrinolytic system. Growth-arrested SMC released active PAI
(101 arbitrary units (AU)/10(6) cells/24 h) and a latent form of PAI (880
AU/10(6) cells/24 h) into the conditioned medium (CM). The levels of PAI
were significant since 880 AU of PAI could inhibit approximately 1
microgram of tissue plasminogen activator. The extracellular matrix of SMC
also contained PAI activity; however, the level was 17-fold less than that
observed in the CM. SMC-PAI was a rapid inhibitor of tissue plasminogen
activator (kass greater than 10(7) M-1 S-1) and was identified as a 45-kDa
protein immunologically related to endothelial cell PAI-1. PAI-1 comprised
20 and 30%, respectively, of the newly synthesized protein detected in the
CM and extracellular matrix of SMC. The SMC growth modulators, platelet-
derived growth factor and transforming growth factor-beta, induced PAI- 1
activity and protein synthesis by 2- and 3-fold, respectively, in a dose-
and time-dependent manner. The increases in PAI-1 activity and protein
synthesis were ascribed to elevated levels of PAI-1 mRNA as judged by
Northern blot analysis of total RNA prepared from control and
platelet-derived growth factor- and transforming growth factor-beta-
treated cells. Increases in PAI-1 mRNA levels were evident 1 h after growth
factor treatment and were maximal after 4 h. PAI-1 mRNA levels were
unaffected by cycloheximide treatment. The results indicate that SMC
synthesize and release PAI-1 which could regulate the normal fibrinolytic
environment of the arterial wall. During atherosclerosis or after vascular
injury increases in platelet-derived or locally produced mitogens may
stimulate further PAI-1 synthesis and generate a prothrombotic state.
Platelet-derived growth factor and transforming growth factor-beta regulate plasminogen activator inhibitor-1 synthesis in vascular smooth muscle cells
Department of Pharmacology, Merck Sharp and Dohme Research Laboratories, West Point, Pennsylvania 19486.
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