Transformed rat arterial smooth muscle cells induce platelet aggregation.
Atherogenesis is characterized by a proliferation of arterial smooth muscle cells that may be of transformed nature. Platelets are implicated in the progression of atherosclerotic lesions through thrombotic complications. The present study was designed to investigate whether transformed arterial smooth muscle cells (SMC) could specifically aggregate platelets. We used rat transformed arterial SMC lines, V6- and V8-lines, that we had previously established. Experiments were performed with an in vitro homologous rat system. Suspensions of SMC were added without any other aggregating agent to rat heparinized platelet-rich plasma (PRP) in a coagulo-aggregometer. The effect of transformed V6-line and V8-line SMC was compared to that of their normal parental counterparts, V6- and V8-parent cells. Suspensions of transformed SMC induced, in a dose-dependent manner, an immediate and reversible ADP-like platelet aggregation. The amplitude of platelet aggregation was much higher with addition of transformed cells than of the corresponding control SMC (7.39 +/- 0.75 cm vs. 0.85 +/- 0.62 cm with 2 x 10(6) SMC, V6-line vs. V6-parent cells, respectively). ADP-like aggregation did not significantly differ between the two transformed V6- and V8-lines. ADP-like platelet aggregation was also obtained with supernatants of transformed SMC suspensions, the amplitude being higher with supernatants than with cell suspensions (21.0 +/- 3.64 cm vs. 6.8 +/- 1.22 cm with 1.0 x 10(6) V8-line cells, supernatant vs. cell suspension, respectively). The transformed SMC-induced aggregation of platelets was inhibited by apyrase (125 microM) and iodoacetate (25 mM) and thus was ascribable to ADP released by the SMC. In addition, all suspensions of SMC, normal or transformed, but not their supernatants, induced plasma clotting after variable coagulation times. Coagulation was inhibited by hirudin (25 to 100 U/ml) and phospholipase A2 (10 U/ml) indicating thrombin generation through activity of the SMC membrane tissue factor. The present results show that transformed arterial smooth muscle cells may directly aggregate platelets via a release of ADP and this could be of pathophysiological relevance for thrombosis associated with atherosclerosis.
INSERM U63, Lyon-Bron, France.
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N
Blaes
C
C
Covacho
D000244:Adenosine Diphosphate / Q000494:pharmacology; D000818:Animals; D001081:Apyrase / Q000494:pharmacology; D001777:Blood Coagulation; D002461:Cell Line, Transformed; D004492:Edetic Acid / Q000494:pharmacology; D006629:Hirudins / Q000494:pharmacology; D007461:Iodoacetates / Q000494:pharmacology; D019807:Iodoacetic Acid; D009131:Muscle, Smooth, Vascular / Q000502:physiology*; D010739:Phospholipase D / Q000494:pharmacology; D010741:Phospholipases A / Q000494:pharmacology; D054467:Phospholipases A2; D010974:Platelet Aggregation / Q000187:drug effects / Q000502:physiology*; D051381:Rats
D006629:Hirudins; D007461:Iodoacetates; D000244:Adenosine Diphosphate; D004492:Edetic Acid; D010741:Phospholipases A; D054467:Phospholipases A2; D010739:Phospholipase D; D001081:Apyrase; D019807:Iodoacetic Acid