Investigation of the effects of phospholipase C on human platelets: evidence that aggregation induced by phospholipase C is independent of prostaglandin generation, released ADP and is modulated by cyclic AMP.
Effects and the mechanism of action of phospholipase C (PLC), from Clostridium perfringens, on washed human platelets were examined to better understand the role of PLC in platelet function. PLC caused aggregation and secretion of [14C]-5HT, without concomitant loss of cytoplasmic, LDH, in a concentration dependent manner. P-nitrophenylphosphorylcholine, a substrate for PLC, blocked these responses in a concentration dependent manner. In other experiments hirudin, alpha-1-antitrypsin and soybean trypsin inhibitor did not inhibit PLC-induced activation of human platelets. PLC-induced aggregation and [14C]-5HT secretion was not inhibited by aspirin, a known inhibitor of prostaglandin biosynthesis. PLC-induced aggregation was selectively inhibited by analogs of 7,8-dihydroxybenzazepine and 7,8-methylenedioxybenzazepine in a concentration dependent manner. These two agents had no effect on arachidonic acid-induced aggregation. PLC-induced aggregation was not inhibited by apyrase, an enzyme which hydrolyzes ADP. In other experiments, PLC-treated platelets did not exhibit any platelet activating factor-like activity. Prostaglandin E1 and trifluoperazine showed concentration dependent inhibitor effects on PLC-mediated aggregation and secretion of [14C]-5HT. These findings indicate that: a) PLC is capable of inducing aggregation and specific secretion of [14C]-5HT without causing lysis of platelets; b) mechanism of PLC-induced activation of platelets is independent of prostaglandin generation or action, released ADP, and PAF; and c) cyclic AMP plays a modulatory role in PLC-mediated secretion and aggregation of human platelets.
D000244:Adenosine Diphosphate / Q000097:blood*; D001792:Blood Platelets / Q000201:enzymology; D000242:Cyclic AMP / Q000097:blood*; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000097:blood; D010740:Phospholipases / Q000494:pharmacology*; D010974:Platelet Aggregation / Q000187:drug effects*; D011453:Prostaglandins / Q000097:blood*; D012701:Serotonin / Q000378:metabolism; D010738:Type C Phospholipases / Q000494:pharmacology*
D011453:Prostaglandins; D012701:Serotonin; D000244:Adenosine Diphosphate; D000242:Cyclic AMP; D007770:L-Lactate Dehydrogenase; D010740:Phospholipases; D010738:Type C Phospholipases