Krishnamurthi S, Kakkar V V
Thromb Haemost. 1985 Jun 24;53(3):337-42.
The effect of pyridoxal 5'-phosphate (PALP) and trifluoperazine (TFPZ), the calmodulin antagonist, on in vitro platelet adhesion to collagen and collagen-induced platelet activation was studied using platelet-rich-plasma (PRP) or washed platelets (WPL). Platelet aggregation and [14C]-5HT release induced by "threshold" or low concentrations of collagen (0.6 micrograms/ml) in PRP were completely abolished by PALP (24 mM), TFPZ (250 microM) as well as indomethacin (10 microM). At higher concentrations of collagen (10-15 micrograms/ml) in PRP and WPL, the use of stirred and unstirred platelets treated with collagen enabled a distinction to be made between aggregation and adhesion-mediated release reaction. Platelet aggregation and the aggregation-mediated release reaction induced by these concentrations of collagen in stirred platelets were completely abolished by PALP, TFPZ and indomethacin although neither adhesion to collagen nor the adhesion-mediated release reaction of unstirred platelets was significantly affected by these inhibitors. Interestingly, both adhesion and the adhesion-mediated release reaction were abolished by concentrations of PALP 10-40 fold higher than those required to abolish aggregation. Collagen-induced platelet aggregation, but not platelet adhesion, was inhibited in resuspended platelets pretreated with PALP and NaBH4 indicating a separation in the membrane sites involved in aggregation and adhesion. The results further emphasize the distinction between adhesion and aggregation-mediated events with regards to collagen with the latter being more susceptible to inhibition by antiplatelet agents such as PALP and TFPZ.
利用富含血小板血浆(PRP)或洗涤血小板(WPL),研究了5'-磷酸吡哆醛(PALP)和钙调蛋白拮抗剂三氟拉嗪(TFPZ)对体外血小板与胶原黏附及胶原诱导的血小板活化的影响。PRP中由“阈值”或低浓度胶原(0.6微克/毫升)诱导的血小板聚集和[14C]-5羟色胺(5HT)释放,被PALP(24毫摩尔)、TFPZ(250微摩尔)以及吲哚美辛(10微摩尔)完全抑制。在PRP和WPL中较高浓度胶原(10 - 15微克/毫升)时,对经胶原处理的搅拌血小板和未搅拌血小板进行比较,能够区分聚集和黏附介导的释放反应。这些浓度的胶原在搅拌血小板中诱导的血小板聚集和聚集介导的释放反应,被PALP、TFPZ和吲哚美辛完全抑制,尽管这些抑制剂对未搅拌血小板与胶原的黏附以及黏附介导的释放反应均无显著影响。有趣的是,能消除聚集所需浓度10 - 40倍的PALP浓度可消除黏附及黏附介导的释放反应。用PALP和硼氢化钠预处理后再悬浮的血小板中,胶原诱导的血小板聚集受到抑制,但血小板黏附未受抑制,这表明参与聚集和黏附的膜位点是分开的。结果进一步强调了在与胶原相关的黏附事件和聚集介导事件之间的区别,后者更易受到PALP和TFPZ等抗血小板药物的抑制。