奥卡宁的提取分离及抗血小板聚集作用研究
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摘要
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 of 900μg/mL,medium dose of 300μg/mL,low dose of 100μg/mL),okanin group (high dose of 45μg/mL,medium dose of 15μg/mL,low dose of 5f 15 igh dose of 45 mly 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.
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 of 900μg/mL,medium dose of 300μg/mL,low dose of 100μg/mL),okanin group (high dose of 45μg/mL,medium dose of 15μg/mL,low dose of 5f 15 igh dose of 45 mly 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.
引文
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[2]刘立业.HPLC-ELSD法测定昆仑雪菊中氨基酸种类[J].食品研究与开发,2016,37(17):118-120.
[3]刘金俊.两色金鸡菊头状花序的化学成分和药材质量研究[D].北京:北京协和医学院,2015.
[4]DIAS T,BRONZE M R,HOUGHTON P J,et al.The flavonoid-rich fraction of coreopsis tinctoria promotes glucose tolerance regain through pancreatic function recovery in streptozotocin-induced glucose-intolerant rats[J].Journal of Ethnopharmacol,2010,132(2):483-490.
[5]茹克娅·胡加,阿不都拉,斯依提,尼沙汗·吾满尔.雪菊花药用功能及临床应用体会[J].中国民族民间医药,2011,20(19):41-42.
[6]卢伟,马晓丽,兰怡,等.两色金鸡菊醇提物对自发性高血压大鼠尿液的代谢组学研究[J].中国药理学通报,2014,29(9):1311-1314,1315.
[7]梁淑红,庞市宾,刘晓燕,等.金鸡菊提取物降血脂作用的研究[J].中国实验方剂学杂志,2010,16(8):234-235.
[8]王健.两色金鸡菊提取物大鼠肠吸收及预防血栓作用与物质基础的研究[D].乌鲁木齐:新疆医科大学,2016.
[9]ZHAO L M,JIN H S,SUN L P,et al.Synthesis and evaluation of antiplatelet activity of trihydroxychalcone derivatives[J].Bioorganic and Medicinal Chemistry Letters,2005,15(22):5027-5029.
[10]黄敏珠,陈海生,刘建国,等.中药鬼针草化学成分的研究[J].第二军医大学学报,2006,26(8):888-891.
[11]张川.银杏达莫注射液联合前列地尔在急性脑梗死急诊治疗中对患者血液流变学、血小板活性指标和神经功能的影响[J].川北医学院学报,2018,33(4):483-487.
[12]CALONGE N,PETITTI D B,DEWITT T G,et al.Aspirin for the prevention of cardiovascular disease:U.S.preventive services task force recommendation statement[J].Ann Intern Med,2009,150(6):396-404.
[13]王炎炎,朱惠超,肖扬,等.注射用血栓通体外对家兔血小板聚集的影响[J].中草药,2014,45(18):2669-2672.
[14]DORSAM R T,KUNAPULI S P.Central role of the P2Y12receptor in platelet activation[J].J Cin Invest,2004,113(3):340-345.
[15]史帝,崔桂云.凝血酶在脑出血中作用机制的研究[J].中国实用神经疾病杂志,2014,17(10):138-140.