壳聚糖体外促进ADP诱导人血小板聚集作用及其机制

The Effect and Mechanism of Chitosan on Human Platelet Aggregation Induced by ADP in Vitro

为了观察壳聚糖溶液在有无诱导剂二磷酸腺苷(ADP)存在的条件下对人血小板聚集的影响,通过体外聚集实验,用比浊法检测富含血小板的血浆(PRP)及洗涤后血小板的聚集率,用ELISA的方法对人血小板聚集后释放的产物P-选择素(P-selectin)、血栓素B_2(Thromboxane B_2,TXB_2)进行含量测定;加入低剂量的ADP进行诱导并用抗体SZ-21对ADP途径进行封闭,从而确定壳聚糖是否通过ADP途径引起血小板的聚集。结果显示,在无诱导剂的情况下,壳聚糖能够引起血小板的聚集(P<0.01),聚集后的产物能释放出大量的活性物质P-selectin、TXB_2(P<0.01);加入低剂量诱导剂ADP后,聚集效果更加明显(P<0.01),封闭ADP途径后,加入壳聚糖溶液不能够引起血小板聚集。以上实验结果提示壳聚糖能够促进血小板的聚集,并释放出活性物质,且与ADP起协同作用,说明壳聚糖引起血小板聚集是通过ADP途径起作用的,这也可能是其止血的机理之一。

In this study, the effect of chitosan on human platelet aggregation with or without inducer ADP was investigated. Through the given drugs in vitro, the aggregation of the platelet rich plasma （PRP） and the washed platelet was deteted by the nephelometric method, and the concentration of active substances, such as Pselectin and Thromboxane B2 were also measured by the ELISA. Then through adding the low-dose ADP and using the antibody SZ- 21 ADP pathway was blocked out. The purpose is to make sure that if chitosan can cause platelet aggregation through ADP pathway. Without ADP, Chitosan can promote the PRP＇s aggregation, but platelet aggregation level was lower than that of ADP. The products of aggregation can also release a large number of active substances such as P-selectin and Thromboxane B2 （P 〈 0. 01）. After blocking ADP pathway, adding the chitosan can not promote the platelet aggregation. Chitosan can promote the platelet aggregation and cooperate with ADP, and increase the active moleculars released from the platelet. It showed that chitosan caused platelet aggregation through ADP pathway.