薄層クロマトグラフィーによるヒト血小板アラキドン酸代謝の解析
Analysis of arachidonic acid metabolism by human platelets using thin layer chromatography (TLC)

Kazuo KOIKE, Haruo ANDO, Hideo SASAKI, Holm HOLMSEN, A. Koneti RAO
1982 Blood & Vessel  
Metabolism of arachidonic acid (AA) by human platelets was studied by TLC. Gelfiltered platelets (GFP) were eluted both with Ca2+-free Tyrode albumin solution (pH 7. 3) and with albumin-free Haslam solution (pH 7.4). GFP was stimulated by different concentrations of thrombin (f.c.: 0.06-5u/ml). Platelet lipids were extracted by the method of Lapetina and Cuatrecasas, followed by TLC in the two kinds of slovent systems : I; Tou's solvent system; chloroform/methanol/acetic acid/H20
more » ... /H20 (81:10:45:5v/v) and II: the solvent system of Hong and Levine; the upper layer of ethylacetate/glacial acetic acid/iso-octane/ H2O (90:20:50:100, v/v). 14C-AA was incorporated into platelet phospholipids: PC 60.7%, PI 16.9%, PE 10.8% and PS 10.2%. On the other hand, free AA was released mainly from PC and PI. Radioactivity of 13 % of free 14C-AA was liberated from phospholipids after stimulation of GFP with 5u/ml of thrombin for 5 min. AA metabolites, such as PGD2, PGE2, TXB2f HETE and HHT were also detected by TLC. No spots and radioactivities corresponding to HHT were detected in aspirin (1 mM) -treated normal platelets and in platelets from the patient with congenital cyclooxygenase deficiency after stimulation by thrombin. Free AA was not liberated from phospholipids in platelets from the patient with congenital phospholipase A2 deficiency.
doi:10.2491/jjsth1970.13.163 fatcat:uyweel3r2ba3ncin2ik6gh3nqq