莪术二酮抑制凝血酶诱导血小板活化和聚集的研究  

Inhibition by curdione of thrombin-induced platelet activation and aggregation

作　　者：乔文豪 张冬玲[1] 赵营莉 黄燕[2] 孙继敏[2] 许杜娟[1,2] 夏泉[1,2] 

机构地区：[1]安徽医科大学药学院,合肥230032 [2]安徽医科大学第一附属医院药剂科,合肥230022

出　　处：《安徽医科大学学报》2017年第3期376-382,共7页Acta Universitatis Medicinalis Anhui

基　　金：安徽省自然科学基金(编号:1408085MH172)

摘　　要：目的研究莪术二酮对凝血酶诱导大鼠血小板活化和聚集的影响,来阐明莪术二酮抗血小板活化的机制。方法电阻抗法测定凝血酶诱导大鼠洗涤血小板聚集率,计算莪术二酮对其抑制率。荧光分光光度法测莪术二酮抑制血小板胞内钙离子浓度变化。流式细胞术检测血小板P-selectin(CD62p)的表达。Western blot测定磷酸化蛋白的磷酯酶C(PLC)、蛋白激酶C(PKC)和丝裂原活化蛋白激酶(MAPKs)的变化。结果 400μmol/L的莪术二酮可以有效地抑制0.3 U/ml凝血酶诱导的大鼠血小板的聚集。50、100μmol/L的莪术二酮可以显著抑制血小板活化标志物[Ca^(2+)]i上升和P-selectin(CD62p)的表达。在蛋白水平上,莪术二酮可以抑制PLCβ3、PKCθ和MAPKs蛋白的磷酸化。结论初步认为莪术二酮通过PLC-PKC-MAPKs通路抑制凝血酶诱导的血小板活化和聚集,阐明了莪术二酮抗血小板活化的部分机制。Objective To clarify the mechanism of curdione-attenuated platelet activation through exploring the effects of curdione on thrombin - induced rat platelet activation and aggregation. Methods Platelet aggregation was measured with the electrical impedance method then the inhibition rate of curdione was calculated. Inhibitory effect of curdione on intracellular calcium concentration in platelets was detected by fluorescence spectrophotometry. The expression of P-selectin（CD62p） was measured by flow cytometry. The phosphorylation of PLC, PKC and MAPK proteins were detected by Western blot. Results The results showed that 0. 3 U/ml thrombin-induced rat platelet aggregation was inhibited by 400 μmol/L eurdione effectively. The markers of platelet activation included the up - regulation of intracellular calcium mobilization [ Ca2 ＋ ]i and the expression of P-seleetin （CD62p） were decreased by 50 μmol/L or 100 μmol/L curdione significantly. Meanwhile, Western blot results demonstrated that the phos- phorylation of the PLCβ3, PKC0 and MAPKs was also inhibited by curdione. Conclusion Curdione may inhibit thrombin-induced rat platelet activation and aggregation by inhibiting the PLC-PKC-MAPKs pathway, at least in part.

关 键 词：莪术二酮 凝血酶 血小板活化和聚集 PLC-PKC-MAPKs通路 

分 类 号：R285.5[医药卫生—中药学] R331.1[医药卫生—中医学]