[Inducing Effect of PKA Inhibitor at Different Concentrations on Platelet Apoptosis and Its Mechanism].

To investigate the inducing effect of PKA inhibitor H89 of different concentrations on platelet apoptosis and its mechanism.Platelets were isolated from peripheral venous blood of healthy volunteers. Different concentrations gradient PKA inhibitor H89 were co-incubated with washing platelets, and the effects of PKA inhibitor H89 at different concentrations on platelet mitochondrial membrane potential, phosphatidylserine and reactive oxygen species(ROS) were determined by ELISA and flow cytometry.Different concentration of PKA inhibitor H89 could induce the depolarization of mitochondrial membrane potential and PS exposure of platelet. However, high concentration(100 μmol/L) PKA inhibitor H89 could induce the production of ROS in platelets, but medium and low concentrations did not induce the production of ROS in platelets. And several ROS inhibitors could inhibit the apoptosis induced by high concentration PKA inhibitor H89.High concentration H89 can induce platelet apoptosis, however the mechanism of platelet apoptosis caused by H89 of high concentration is different from H89 at medium and low concentrations.不同浓度PKA抑制剂对血小板凋亡的诱导作用及机制.探讨不同浓度PKA抑制剂H89对血小板凋亡的诱导作用和机制.取健康志愿者外周静脉血分离血小板，选择不同浓度梯度的PKA抑制剂H89与经洗涤的血小板共同孵育，应用酶联免疫吸附试验、流式细胞术等检测不同浓度PKA抑制剂H89对血小板线粒体膜电位（Mitochondrial membrane potential，Δψm）、磷脂酰丝氨酸（Phosphatidylserine，PS）和活性氧（Reactive oxygen species，ROS）的影响.不同浓度PKA抑制剂H89均能诱导血小板发生Δψm去极化和PS暴露，但高浓度（100 μmol/L）PKA抑制剂H89可诱导血小板产生ROS，而中低浓度则不会诱导血小板内ROS的产生，并且多种ROS抑制剂均可抑制高浓度H89诱导的凋亡.高浓度的PKA抑制剂H89能诱导血小板凋亡，但是作用机制不同于中低浓度.