韭菜的抗凝血活性及其成分分析

Analysis of the Anticoagulant Activity and Ingredients of Chives

目的:为弄清中医理论上韭菜具有活血和止血的原因,以及了解食用韭菜是否可能对凝血不稳定或正服用抗凝血药剂的人群产生影响,对韭菜进行体外抗凝血试验。方法:干韭菜用70%乙醇60℃浸提,所得浸膏依次用石油醚、乙酸乙酯、正丁醇、水进行分相萃取。水相用乙醇沉淀分离。将乙醇沉淀的上清液过活性炭柱,收集不同浓度的乙醇洗脱液。95%乙醇洗脱液用Sephadex LH-20分离,收集洗脱液。对各种分离物质进行体外抗凝血试验。对Sephadex LH-20分离的具有较强抗凝血活性6个组分用多种显色技术进行物质类别鉴定。结果:体外抗凝血研究中,正丁醇的抗凝血作用极强。韭菜中正丁醇萃取相能明显降低该作用,说明正丁醇萃取相中含有促进血浆凝集物质;水萃取相能够显著延迟凝血时间。韭菜乙醇沉淀上清液和沉淀均能抑制凝血,乙醇沉淀上清液抗凝血效果强。韭菜乙醇沉淀上清液过活性炭柱后,其95%乙醇洗脱液抗凝血活性最强。该洗脱液过Sephadex LH-20柱后得到的8个分离相,有6个具有较强的抗凝血活性,其中S2抗凝血活性最强。该6种物质属于有机物,即4种酸性,不含有糖基;2种p H中性,含有糖基,在365 nm处显示为蓝色荧光,没有明显的香豆素的开环闭环反应,均带有氨基。结论:韭菜水溶性物质具有强烈的抗凝血作用,而其正丁醇提取物具有促进凝血作用。水相中的主要抗凝血成分是一大类物质,可能为氨基-苯丙酸类及其苷类物质。

Objective： In order to understand the reasons of traditional Chinese medicine about chives（Allium tuberosum） with invigorate the circulation of blood and hemostasis, and find out if chives leaf affect on coagulant excursion or taking anticoagulant medicine people, chives leaf in vitro anti-coagulation experiments were carried out. Methods： Dried chives leaf were extracted with 70% ethanol at 60 ℃, and its extraction were sub-phase extracted by petroleum ether,ethyl acetate, butanol and water in turn. The water phase was precipitated and separated with alcohol. Alcohol precipitation supernatant were separated by activated carbon column, and different concentrations of ethanol eluate were collected.95% ethanol eluate was separated with Sephadex LH-20 and the eluate were collected. A variety of separate substances were experimented with vitro anti-coagulation. The six components with strong anticoagulant activity after the separation by Sephadex LH-20 were identified by a variety of chromogenic reaction technology. Results： The results showed butanol had strong anticoagulant effect in vitro anticoagulant. Chives leaf butanol extraction phase significantly reduced anticoagulant effect that indicates this phase contains promoting plasma agglutination substances; water extraction phase significantly delayed the clotting time. Chives leaf alcohol precipitation and supernatant could inhibit blood coagulation, and supernatant had a stronger anticoagulant effect. 95% ethanol eluate of the supernatant separated by activated carbon column was strongest anticoagulant activity between all eluate. Eight separation phases obtained by Sephadex LH-20 column, and six substances of them had strong anticoagulant activity, including S2 with the strongest anticoagulant activity. The six substances are organic matter, four kinds of them are acidic and not contain glycosylation, two kinds of them are neutral p H and containing glycosylation that show blue fluorescent under 365 nm, but without coumarin ring closing reaction, all six substances have amino groups. Conclusion： Chives leaf water-soluble substances have a strong anticoagulant effect,while the butanol extract promotes the coagulation of the blood. Anticoagulant in the aqueous phase are a large class of substances that may be amino-styrene-acrylic acids and their glycosides.