奥卡宁的提取分离及抗血小板聚集作用研究

Extraction and separation of okanin and its exploration on antiplatelet aggregation effect

目的分离和纯化两色金鸡菊中的奥卡宁,并研究其抗血小板聚集作用。方法两色金鸡菊干燥花序经乙醇回流提取得到两色金鸡菊醇提物(Ethanol Extract,ETE),用乙酸乙酯和正己烷对ETE水溶液进行萃取,采用硅胶中压柱层析方法对萃取物进行分离得到奥卡宁;采集静脉血,随机分为空白组(生理盐水)、ETE高、中、低(900、300、100μg/mL)剂量组、奥卡宁组高、中、低(45、15、5μg/mL)剂量组、阿司匹林(acetylsalicylic acidsigma,ASA)组(500μg/mL),建立二磷酸腺苷(adenosine diphosphate,ADP)、凝血酶诱导的血小板聚集模型,采用Born比浊法,以血小板聚集抑制率为指标,观察其抗血小板聚集作用。结果从两色金鸡菊中分离得到奥卡宁。与空白组比较,ETE中、高剂量组和奥卡宁高剂量组可以抑制ADP诱导的血小板聚集,差异有统计学意义(P <0.01)。与空白组比较,ETE中剂量组和奥卡宁中剂量组可以抑制凝血酶诱导的血小板聚集,差异有统计学意义(P <0.05),ETE高剂量组和奥卡宁高剂量组可以抑制凝血酶诱导的血小板聚集(P <0.01)。结论两色金鸡菊中的奥卡宁是抗血小板聚集的主要活性物质。

Objective To isolate and purify okanin from coreopsis tinctoria and to test its anticoagulant effect.Methods The dried inflorescence of coreopsis tinctoria was refluxed with ethanol to obtain ethanol extract(ETE).The ETE aqueous solution was extracted with ethyl acetate and n-hexane.The extract was separated by silica gel column chromatography to obtain okanin.Venous blood was collected and randomly divided into blank group(saline),ETE group(high dose of900μg/mL,medium dose of300μg/mL,low dose of100μg/mL),okanin group(high dose of45μg/mL,medium dose of15μg/mL,low dose of5f15igh dose of45mly divided into blank group(saline),ETEμg/mL),establish adenosine diphosphate(ADP),thrombin-induced platelet aggregation model,Born turbidimetry was used to observe the platelet aggregation inhibition rate and observe its anti-platelet aggregation effect.Results Okanin was isolated from the coreopsis tinctoria.Compared with the blank group,the medium and high dose groups of ETE and the high dose group of okanin could inhibit ADP-induced platelet aggregation(P<0.01).Compared with the blank group,the medium dose group of ETE and the medium dose group of okanin could inhibit thrombin-induced platelet aggregation(P<0.05).The high dose of ETE group and the high dose of okanin group could inhibit thrombin-induced platelets aggregation(P<0.01).Conclusion Okanin in the coreopsis tinctoria is the main active substance for anti-platelet aggregation.