荜茇酰胺对儿童抗凝、抗血小板聚集作用的体外实验研究

Inhibition of piperlongumine on platelet aggregation and blood coagulation for children in vitro study

目的观察荜茇提取物荜茇酰胺(PL)对儿童具有抗血小板、抗凝作用,为临床用药提供实验依据。方法抽取30名健康儿童静脉血,随机分为5组,分离富血小板血浆(PRP),取PRP分别加入二甲基亚砜(DMSO)作为空白组,加入阿司匹林(终浓度10μmol/L)作为对照组,加入PL(终浓度分别为20、100、200μmol/L)作为不同浓度PL组,37℃孵育5min,作用后PRP分别加入诱导剂二磷酸腺苷(ADP)终浓度10μmol/L,胶原(Coll)终浓度2.5μg/mL,花生四烯酸(AA)500μg/mL,采用比浊法在37℃条件下测定血小板聚集率。抽取静脉血分离血浆后分5组,将不同浓度PL分别与3组血浆混合,终浓度分别为5、10、20μmol/L,空白组将1%DMSO与血浆混合,对照组将肝素钠与血浆混合(35U/mL),各组37℃孵育5min后上机检测纤维蛋白原(FIB)水平、凝血酶原时间(PT)及凝血酶时间(TT)、活化部分凝血酶原时间(APTT)。结果与对照组比较,PL(20、100、200μmol/L)干预后对AA、Coll诱导的血小板聚集均有显著抑制作用(P<0.05),PL 100μmol/L、PL 200μmol/L可以显著抑制ADP诱导的血小板聚集率(P<0.05);PL 10μmol/L、PL 20μmol/L干预后可以显著延长儿童血浆的PT、APTT、TT值(P<0.05),但FIB未见明显变化。结论 PL能抑制儿童血浆的血小板聚集,抑制抗凝活性。

Objective To observe the inhibition of piperlongumine in vitro on platelet aggregation and blood coagulation tests for children,to provide the experimental basis for clinical medication.Methods Venous blood samples from 30 children were randomly devided into 5groups,and was centrifuge to separate platelet-rich plasma(PRP).After storing in 37℃thermostat water bath for 5minutes,the PRP which have been added DMSO as blank group,and added Aspirin(10μmol/L)as control group,and added PL(20μmol/L),PL(100μmol/L),PL(200μmol/L)as different concentrations of PL groups respectively,were induced by the addition of adenosine diphosphate(10μmol/L),collagen(2.5μg/mL)and the arachidonic acid(500μg/mL).Then the platelet aggregation rate of the PRP from 5groups could be measured by turbidimetry.Blood plasma isolated from venous blood was divided into 5groups.In the PL groups,blood plasma were mixed up with PRP concentrations of which were 5,10,20μmol/L.In the bland group,blood plasma were mixed up with DMSO(1%).In the control group,blood plasma were mixed up with heparin sodium(35U/mL).After storing in 37 ℃thermostat water bath for 5minutes,fibrinogen(FIB),prothrombin time(PT),thrombin time(TT)and activated partial thromboplastin time of different groups were detected.Results Compared to the control group,the groups which were add PL with different concentrations(20,100,200μmol/L)showed significant inhibition on platelet aggregation induced by AA and collagen(P<0.05).PL with concentrations of 100μmol/L and 200μmol/L showed significant inhibition on platelet aggregation induced by ADP(P<0.05).The PT,APTT,TT of blood plasma from children had been significantly prolonged by the intervention of PL 10μmol/L and PL 20μmol/L(P<0.05),however,no significant change of FIB was observed.Conclusion There are inhibitory effects of PL on platelet aggregation of blood plasma from children and anticoagulant activity in this study.