海星中抑制ACE活性的甾体化合物的分离纯化  

Separation and purification of steroids with inhibitory activity of angiotensin I-converting enzyme in starfish Asterias amurensis

作　　者：田丽冉 朱春芃 曲敏[1] 佟长青[1] 金桥[1] 谭成玉[2] 李伟[1] 

机构地区：[1]大连海洋大学食品科学与工程学院,辽宁大连116023 [2]大连海洋大学海洋科技与环境学院,辽宁大连116023

出　　处：《大连海洋大学学报》2014年第4期409-412,共4页Journal of Dalian Ocean University

基　　金：国家海洋局公益性行业科研专项(201205022-7);大连市科学技术基金资助项目(2011J21DW012)

摘　　要：用体积分数为70%的乙醇对多棘海星Asterias amurensis进行热浸提，减压浓缩后依次用石油醚、乙酸乙酯、正丁醇进行萃取，分别浓缩乙酸乙酯层和正丁醇层，得到海星总甾体化合物，对总甾体化合物的含量和抑制血管紧张素Ⅰ转换酶（ ACE）的活性进行了测定，并将海星总甾体化合物通过正向硅胶柱色谱和Sephadex TM LH-20柱色谱进一步分离纯化，根据薄层色谱分析收集有效成分。结果表明：正丁醇层中总甾体化合物的含量最高，石油醚层中的含量最低；海星乙醇粗提物、乙酸乙酯层和正丁醇层中的总甾体化合物对ACE酶活性有明显的抑制作用，而石油醚粗提物的抑制作用则较低，其中浓度为100 mg/mL的正丁醇层溶液对ACE酶的抑制率最高（78.05%），其IC50值最低，为27.76 mg/mL；正丁醇层中的总甾体化合物通过正向硅胶和Sephadex LH-20柱色谱得到F1-F88个亚组分，其中F3在薄层色谱（ TLC）中为单点，醋酸酐-浓硫酸反应（ Liebermann-Burchard反应）为阳性，提示为甾体类化合物。The total steroids were extracted from starfish Asterias amurensis by 70% ethanol, and then by petroleum ether, ethyl acetate and n-butyl alcohol, and the ethyl acetate and n-butyl alcohol extracts were evaporated. The inhibitory activity of angiotensinⅠ-coverting enzyme（ ACE） in total steroids was determined and purified by differ-ent chromatographic methods including silica gel column and Sephadex TM LH-20 for screening of bioactive constitu-ents from the starfish. The determination showed that there was the maximal steroids in the n-butyl alcohol extracts and the minimal steroids in petroleum ether extracts. The ethanol extracts, ethyl acetate and n-butyl alcohol ex-tracts were shown to contain inhibitory activity of ACE, the maximal ACE inhibition rate （78. 05% at 100 mg/mL） and minimal IC50（27. 76 mg/mL） in the n-butyl alcohol fraction, and lower ACE inhibition in the petroleum frac-tion. Eight subunits from F1 to F8 were obtained by silica gel column and Sephadex LH-20, in which only F3 with positive L-B reaction showed single spot by thin-layer chromatogram（ TLC） .

关 键 词：海星 甾体化合物 血管紧张素Ⅰ转换酶(ACE) 

分 类 号：R914[医药卫生—药物化学]