Human platelets release a paf-acether : acetylhydrolase similar to that in plasma

Intact washed human platelets aggregated in response to paf-acether (paf) and did not metabolize [3H]paf at concentrations up to 10 nM. However, when platelets were lysed by exposure to pH 9.5, resulting in 37.5±2.5% (mean ±SD, n=3) lactic dehydrogenase (LDH) release, 20.5±5.7% of the radioactivity was detected as labeled lyso paf and 5.7±3.1% as labeled alkylacylglycerophosphocholine. When platelets were aggregated with 0.5 IU/mL thrombin or high concentrations of paf (100 nM), they released a part of their acetylhydrolase without releasing LDH. In supernatants obtained from aggregated platelets, 21±2% or 10±2% (n=3), respectively, of the total platelet acetylhydrolase activity was detectedvs. none in supernatants of resting cells. The release of acetylhydrolase was concentration-and time-dependent and paralleled the release of PF 4, a marker for α-granules. The acetylhydrolase affinity for paf (Km) measured in sonicates of resting and thrombin-activated platelets was 8.3±1.5 μMvs. 10.6±1.5 μM, n=5, n.s. in a “Mann Whitney” test. The latter Km was slightly but significantly different (P<0.05, n=5) from that of the thrombin-released acetylhydrolase (7.9 ±1.5 μM) and that of the latter was itself different from plasma acetylhydrolase (5.3±0.5,P<0.05, n=5). Addition of plasma (acid-treated to inactivate acetylhydrolase) decreased the Km value of supernatant acetylhydrolase to 6.1±1.4 μM. All preparations of acetylhydrolase exhibited similar pH requirements and sensitvity to various inhibitors. Thus paf and thrombin cause release of acetylhydrolase from platelets in parallel with release of the α-granule marker PF4. This phenomenon might represent a protective mechanism against paf-mediated effects in thrombotic and cardiovascular diseases.