油枣中黄酮类化合物体外抗凝血活性分析

Analysis of in vitro Anticoagulant Activity of Flavonoids from Youzao Jujube

为了研究油枣中黄酮类化合物的体外抗凝血活性和抗血栓作用,试验以凝血酶原时间(PT)、活化部分凝血酶原时间(APTT)和凝血酶时间(TT)为评价指标,分别测定了油枣中黄酮提取物表儿茶素、槲皮素、异槲皮苷、金丝桃苷、芦丁的体外抗凝血活性;进一步选取抗凝血效果较为明显的黄酮类化合物,进行体外血凝块溶解率试验。结果表明,油枣中5种黄酮类化合物对正常人血浆的APTT均有不同程度的延长,且呈量效关系,其中芦丁的抗凝血效果最明显,当芦丁质量浓度为20 mg/mL时,APTT值达60.8 s,5种黄酮类化合物对APTT的影响由高到低依次为芦丁>表儿茶素>槲皮素>金丝桃苷>异槲皮苷。5种黄酮类化合物对PT和TT略有延长,当芦丁质量浓度为20 mg/mL时,作用于血浆,其PT值为17.8 s;当芦丁质量浓度为40 mg/mL时,作用于血浆,其TT值为23.9 s。当芦丁质量浓度为50 mg/mL,作用于血凝块4、8、24 h后,测得血凝块溶解率分别为22.8%、34.2%、41.3%,显著高于空白对照组。油枣中黄酮类化合物具有体外抗凝血活性和抑制血栓形成的作用。

In order to study the anticoagulant activity and antithrombotic effect of flavonoids in Youzao jujube in vitro,in this study,prothrombin time(PT),activated partial prothrombin time(APTT)and thrombin time(TT)were used as the evaluation indexes to determine the in vitro anticoagulant activity of flavonoid extracts including epicatechin,quercetin,isoquercitrin,hyperin,and rutin from Youzao jujube.The flavonoids with obvious anticoagulant effect were further selected to conduct the clot dissolution rate test in vitro.The results showed that the five flavonoids from Youzao jujube prolonged the APTT of normal human plasma in varying degrees,and showed a dose effect relationship.The anticoagulant effect of rutin was the most obvious.When the mass concentration of rutin was 20 mg/mL,the APTT value reached 60.8 s.The effect of the five flavonoids on APTT from high to low was:rutin>epicatechin>quercetin>hyperin>isoquercetin.They slightly prolonged PT and TT.When mass concentration of rutin was 20 mg/mL,it acted on plasma,and its PT value was 17.8 s.When the mass concentration of rutin was 40 mg/mL,it acted on plasma and its TT value was 23.9 s.The dissolution rates of blood clots were 22.8%,34.2%and 41.3%respectively when the mass concentration of rutin was 50 mg/mL and was applied to blood clots for 4,8,and 24 h,which were significantly higher than those in blank control group.The study above showed that flavonoids in Youzao jujube had anticoagulant activity in vitro and could inhibit thrombosis.