芍药苷联合川芎嗪对肌动蛋白微丝诱导血小板活化及血小板凝溶胶蛋白水平的干预效应研究

目的研究肌动蛋白微丝(F-actin)体外刺激对血小板活化及血小板凝溶胶蛋白(gelsolin)浓度的影响,同时观察活血化瘀中药赤芍-川芎有效成分配伍的体外干预效应。方法以人水洗血小板作为研究对象,分为低(2.5μmol/L)、中(5μmol/L)、高(10μmol/L)浓度F-actin组以及花生四烯酸(AA)、二磷酸腺苷(ADP)、凝血酶(thrombin)组,同时设中药F-actin(10μmol/L)组和中药AA组使用芍药苷和川芎嗪联合干预,阿司匹林F-actin(10μmol/L)组和阿司匹林AA组使用阿司匹林作为干预药物,另设对照组。采用流式细胞术检测血小板活化标志物CD62p的表达,使用血小板聚集仪测定最大血小板聚集率(PAR),采用ELISA法测定血小板gelsolin浓度。结果与对照组比较,不同浓度F-actin及AA、ADP、thrombin均可引起PAR和CD62p表达升高(P<0.01),且F-actin呈浓度依赖性。与AA组比较,低、中浓度F-actin组PAR与CD62p表达下降(P<0.01),高浓度F-actin、ADP、thrombin组则...

Effect of Paeoniflorin and Ligustrazine on the F-actin-induced Platelet Activation Level and Platelet Gelsolin in vitro

Objective To investigate the effect of Paeoniflorin and Ligustrazine on the F-actin-induced platelet aggregation, platelet activation level and platelet gelsolin in vitro. Methods Washing platelets were divided into low(2.5 μmol/L), medium(5 μmol/L) and high(10 μmol/L) F-actin group, AA, ADP and thrombin group. Meanwhile, we took combination of Paeoniflorin and Ligustrazine as the intervention drugs for Chinese medicine F-actin(10 μmol/L) group and Chinese medicine AA group, aspirin for aspirin F-actin(10 μmol/L) group and aspirin AA group. The control group was set up separately. The platelet CD62 p expression was detected by FCM(flow cytometry), platelet aggregation rate(PAR) was detected by platelet aggregation analyzer, and the platelet gelsolin was detected by ELISA. Results Compared with control group, F-actin of different concentrations, AA, ADP and thrombin groups all could increase PAR and CD62 p expression(P<0.01). In addition, F-actin showed concentration-dependent. Compared with AA group, the PAR and CD62 p expression decreased in low and medium F-actin group(P<0.01), while there were no difference in hight F-actin, ADP and thrombin group(P>0.05). Compared with control group, platelet gelsolin increased in medium,high F-actin group and AA group(P<0.05,P<0.01). Compared with AA group,platelet gelsolin decreased in low,medium F-actin group and ADP group,thrombin group(P<0.05). Compared with high F-actin group or AA group, paeoniflorin and ligustrazine, aspirin could inhibit the increasing of PAR and CD62 p expression induced by F-actin or AA(P<0.05). Compared with high F-actin group or AA group, Paeoniflorin and ligustrazine could inhibit the increasing of gelsolin indueced by F-actin or AA(P<0.05), but aspirin had no similar effect(P>0.05). Conclusions F-actin can induce platelet aggregation/activation in vitro, and 10 μmol/L of F-actin can increase the platelet gelsolin level of activated platelet, which has the similar effect with AA. Platelet gelsolin may be an specific effective target for paeoniflorin and ligustrazine against platelet aggregation and activation in vitro.