Determination of the cell lytic properties of amphiphilic inhibitors of the cytosolic phospholipase A2 against human platelets by measuring the liberation of serotonin with high-performance liquid chromatography and fluorescence detection.
A procedure for the determination of the influence of detergents on the cell integrity of human platelets by measuring the release of serotonin with high-performance liquid chromatography and fluorescence detection is presented. After exposure of the platelets to the test compounds the cells and cell fragments, respectively, are centrifuged off. In the supernatants the liberated serotonin is determined directly without a further sample clean up. The amphiphilic inhibitors of cytosolic phospholipase A2 (cPLA2), arachidonyltrifluoromethyl ketone (AACOCF3), palmityltrifluoromethyl ketone (PACOCF3) and methyl arachidonylfluorophosphonate (MAFP), and the polyoxyethylene detergent Brij 58 were investigated for their cell lytic properties with this method. All compounds lysed the platelets liberating serotonin at a concentration of 33 microM. AACOCF, and Brij 58 even caused cell lysis at lower concentrations.
Risse
Dietmar
D
Institute of Pharmaceutical and Medicinal Chemistry, University of Münster, Germany.
Elfringhoff
Alwine Schulze
AS
Lehr
Matthias
M
D001095:Arachidonic Acids / Q000097:blood; D001792:Blood Platelets / Q000201:enzymology*; D002457:Cell Extracts; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003600:Cytosol / Q000201:enzymology*; D004791:Enzyme Inhibitors / Q000097:blood*; D006801:Humans; D006868:Hydrolysis; D007659:Ketones / Q000097:blood; D010741:Phospholipases A / Q000037:antagonists & inhibitors / Q000097:blood*; D054467:Phospholipases A2; D012701:Serotonin / Q000097:blood*; D013050:Spectrometry, Fluorescence
D001095:Arachidonic Acids; D002457:Cell Extracts; D004791:Enzyme Inhibitors; D007659:Ketones; C109341:palmitoyl trifluoromethyl ketone; C081565:arachidonyltrifluoromethane; D012701:Serotonin; D010741:Phospholipases A; D054467:Phospholipases A2