部分蔬菜抗血小板聚集功能及其作用机制的初步研究

Screening of some vegetables that inhibit platelet aggregation and their mechanism： a preliminary study

目的对部分具有抗血小板聚集功能蔬菜的作用机制进行初步探讨。方法将不同种类的蔬菜汁（以西红柿汁作为阳性对照）分别与正常人富血小板血浆（PRP）孵育，加入血小板聚集诱导剂，观察各种蔬菜汁对血小板聚集的影响；用诱导剂激活血小板，流式细胞仪分析蔬菜汁对活化血小板纤维蛋白原受体（Fib-R）和P-选择素（CD62P）的表达水平及血小板纤维蛋白原结合量的影响；利用动物试验对血小板抑制作用进行体内验证，普通白兔随机分为5组，分别定量饲以生理盐水、熟西红柿汁、蒜黄汁、大白菜汁及菠菜汁，观察不同时间后血小板聚集情况。结果在花生四烯酸（AA）诱导下，未发现熟蔬菜汁有明显抑制体外人血小板聚集的作用，但在二磷酸腺苷（ADP）、胶原或肾上腺素诱导下，熟蒜黄汁、大白菜汁、菠菜汁能够明显抑制血小板聚集，并且随着蔬菜汁浓度的增加，对血小板聚集的抑制率增高。熟蒜黄汁、大白菜汁、菠菜汁不能抑制ADP、胶原或肾上腺素诱导的血小板Fib-R、CD62P表达，但能够显著抑制由ADP或胶原诱导的血小板与纤维蛋白原结合量。普通白兔在饲以熟大白菜汁、蒜黄汁、菠菜汁后，ADP诱导的兔血小板聚集率分别在第3、5、8周末时开始降低，熟大白菜汁喂养组血小板聚集的抑制率在第3、5、8周末分别为：45．7％、53．6％、48．5％，熟蒜黄汁喂养组分别为：10．7％、66．7％、46．3％，熟菠菜汁喂养组分别为：8．7％、21．0％、42．6％；饲以熟大白菜汁、蒜黄汁后，胶原诱导的兔血小板聚集率分别在第5、8周末时开始降低，熟大白菜汁喂养组血小板聚集的抑制率在第5、8周末分别为：54．9％、56．3％，熟蒜黄汁喂养组分别为：28．4％、86．7％。在8周内，熟西红柿汁对兔血小板未产生抗聚集作用。结论在ADP或胶原诱导下，熟蒜黄汁、大白菜汁及菠菜汁能明显抑制活化血小板与纤维蛋白原的结合，能够显著抑制血小板的聚集，具有潜在的预防或治疗血栓性疾病的前景。

Objective To study the mechanism of some vegetables inhibiting human platelet aggregation function. Methods Some vegetable juice was mixed （ tomato juice as positive control ） with platelet-rich plasma （PRP） , and maximum ratio of platelet aggregation was measured after induction by agonists on the aggregometer. The expression levels of platelet activation marker were detected, including fibrinogen receptor （Fib-R） , P-selectin （CD62p） , and the combination amount of fibrinogen on the surface of platelet by flow cytometry to validate the inhibitory effect of platelet aggregation by animal tests. Ordinary white rabbits were randomly divided into five groups, each group was fed with saline, cooked tomato juice, blanched garlic leaves juice, Chinese cabbage juice or spinach juice, respectively. And the maximum ratio of rabbit platelet aggregation in different time were observed. Results Cooked vegetable juice couldn＇t inhibit human platelet aggregation induced by AA, but blanched garlic leaves juice, Chinese cabbage juice and spinach juice could inhibit human platelet aggregation obviously induced by ADP, collagen or epinephrine. The inhibition intensity of platelet aggregation increased markedly along with the increase of the concentration of the cooked vegetables juice. Cooked juice of blanched garlic leaves, Chinese cabbage, spinach could not inhibit the expression of Fib-R and CD62p, Whereas they were able to significantly decrease the amount of Fib-R and fibrinogen binding. ADP-induced rabbit platelet aggregation ratios in the group fed with cooked juice of blanched garlic leaves, Chinese cabbage or spinach were significantly lower than control group after 3, 5, 8 weeks, respectively. The inhibition ratios of the platelet aggregation in cooked Chinese cabbage juice group were 45.7% ,53.6% ,48. 5% after 3,5,8 weeks ,respectively. Cooked juice of blanched garlic leaves group were 10. 7%, 66. 7%, 46. 3%, respectively. Cooked spinach juice group were 8. 7%, 21.0% , 42. 6% , respectively. The collagen-induced rabbit platelet aggregation ratios in the groups fed with cooked juice of blanched garlic leaves or Chinese cabbage were significantly lower than control group after 5 and 8 weeks respectively. The inhibition ratios of the platelet aggregation in cooked Chinese cabbage juice group were 54. 9% , 56. 3% after 5 and 8 weeks,respectively. Cooked juice of blanched garlic leaves group were 28. 4% and 86. 7% ,respectively. Cooked tomato juice could not inhibit rabbit platelet aggregation in 8 weeks. Conclusions After induced by ADP or collagen, cooked juice of blanched garlic leaves , Chinese cabbage and spinach could significantly decrease the amount of Fib-R and fibrinogen binding, and inhibit platelet aggregation function significantly. It may have potential application of therapy or prevention of thrombotic diseases.