天麻成分对羟基苯甲醛抗血小板聚集的作用机制研究

Mechanism of Anti-Platelet Aggregation of P-Hydroxybenzaldehyde from Gastrodia Elata

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(P<0.01或P<0.05);不同浓度(320,80,20μmol/L)对羟基苯甲醛对ADP诱导的家兔体外血小板细胞内钙释放与外钙内流均无明显影响,与阴性对照组相比,差异均无统计学意义(P>0.05);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 dual-wavelength Fura-2 fluorescence spectrophotometry. The expression of P2Y（12） receptor induced by ADP was detected by western blotafter 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 P2Y（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 P2Y（12） receptor expression.