天麻成分对羟基苯甲醛抗血小板聚集的作用机制研究

摘要

目的 探讨天麻成分对羟基苯甲醛抗血小板聚集的作用机制.方法 采用酶联免疫吸附法,检测不同浓度(320,80,20μmol/L)对羟基苯甲醛对二磷酸腺苷(ADP)诱导后大鼠体外富血小板血浆中环磷酸腺苷(cAMP)水平的影响;采用双波长Fura-2荧光分光光度法,测定对羟基苯甲醛对ADP诱导后家兔体外血小板胞浆钙离子浓度的影响;采用western blot,检测不同剂量(20,15,10 mg/kg)对羟基苯甲醛大鼠连续灌胃5 d后,对ADP诱导的血小板P2Y12受体表达的影响.结果 对羟基苯甲醛高、中浓度(320,80μmol/L)均能显著提高大鼠富血小板血浆cAMP水平,与模型组比较,差异有统计学意义(P0.05);高剂量(20 mg/kg)对羟基苯甲醛能显著抑制ADP诱导的血小板P2Y12受体的表达.结论 天麻成分对羟基苯甲醛抗血小板聚集的作用是通过提高血小板cAMP水平、抑制血小板P2Y12受体的表达而发挥的.%Objective To investigate the mechanism of the anti-platelet aggregation effect of p-Hydroxybenzaldehyde(PHBA) from Gastrodia elata. Methods Enzyme - linked immunosorbent assay(ELISA)was used to detect the effect of PHBA(320,80,20 μmol/L) on the content of cyclic adenosine monophosphate(cAMP) in platelet-rich plasma(PRP) of rats induced by adenosine diphosphate (ADP). The effect of PHBA on the external platelet cytosolic calcium concentration in rabbits induced by ADP was measured by du-al-wavelength Fura-2 fluorescence spectrophotometry. The expression of P2Y12 receptor induced by ADP was detected by western blot after 5 days of continuous administration of PHBA (20,15,10 mg / kg)rats. Results The high and medium concentrations (320, 80 mol/L) of PHBA can significantly increase the levels of cAMP in PRP of rats,which were significantly higher than that in the model group,and the difference was statistically significant (P < 0. 01 or P < 0. 05). Different concentrations of PHBA (320,80, 20 μmol/L)had no obvious effect on intracellular calcium release and external calcium influx in ADP-induced rabbit platelets. High concentration of PHBA (20 mg / kg ) could significantly inhibit the expression of platelet P 2 Y 12 receptor induced by ADP . Conclusion The mechanism of the anti-platelet aggregation effect of PHBA from Gastrodia elata is related to the increasing of platelet cAMP level and the inhibition of platelet P2Y12 receptor expression.