杨静明¹, 卢虹玉¹, 谢慧风¹, 刘亚月¹, 聂影影^1,2, 张永平¹, 宋采¹, 秦邦伟³, 张翼^1,2
1.广东海洋大学 食品科技学院, 广东省海洋生物制品工程实验室, 广东省水产品加工与安全重点实验室, 广东海洋大学海洋药物研究所, 广东 湛江 524088;2.广东海洋大学 深圳研究院, 广东 深圳 518120;3.山东圣洲海洋生物科技股份有限公司, 山东 威海 264211

摘要:
通过国标方法分析海参酶解原液的基本营养成分；并将海参酶解原液用不同极性的有机溶剂进行提取，利用乙酰胆碱酯酶抑制活性自显影的方法进行活性组分的追踪。利用硅胶柱层析、Sephadex LH-20凝胶柱、高效液相色谱逐步纯化，得到具有较高乙酰胆碱酯酶抑制活性的组分；另外，采用MTT法考察海参酶解液总脂溶性组分对小胶质细胞（BV-2细胞）生长的影响；并利用LPS诱导细胞建立细胞炎症模型，分别采用Griess法和ELISA法检测其对LPS诱导的BV-2细胞中NO和IL-6水平的影响。结果表明：海参酶解原液含有丰富的钙、锌等矿物质，含量较高的多不饱和脂肪酸和必需氨基酸其提取物中具有乙酰胆碱酯酶抑制活性的薄层层析斑点均无明显紫外与荧光吸收，其中活性最强组分C4-2-E在浓度为0.25 g/L时对乙酰胆碱酯酶的抑制率为91.04%；海参酶解液总脂溶性组分在一定的浓度范围内对BV-2细胞均无显著的抑制作用，并且能够显著的抑制LPS诱导的BV-2细胞所产生的NO和IL-6的分泌。故海参酶解液的脂溶性成分可能具有一定的抗神经炎症和老年痴呆作用，具体活性物质的化学结构还需进一步分析鉴定。
关键词:海参提取分离阿尔兹海默氏症乙酰胆碱酯酶抑制活性抗炎活性
DOI：10.11759/hykx20190106001
分类号:TS254.1
基金项目:广东省扬帆计划引进紧缺拔尖人才项目（201433009）；广东省自然科学基金（2018A030307046）；广东海洋大学科研启动项目（E15155）；深圳市科创委基础研究自由探索项目（JCYJ20170306165013264）；深圳市大鹏新区产业发展资金（KY20180203；PT201901-05；KY20170210）

Preliminary study on anti-Alzheimer's activity and composition in sea cucumber hydrolysate
YANG Jing-ming¹, LU Hong-yu¹, Xie Hui-feng¹, LIU Ya-yue¹, NIE Ying-ying^1,2, ZHANG Yong-ping¹, SONG Cai¹, QIN Bang-wei³, ZHANG Yi^1,2
1.College of Food Science and Technology, Guangdong Ocean University, Guangdong Marine Biological Products Engineering Laboratory, Key Laboratory of Guangdong Province Aquatic Products Processing and Safety, Research Institute for Marine Drugs and Nutrition, Guangdong Ocean University, Zhanjiang, 524088, China;2.Shenzhen Insititute of Guangdong Ocean University, Shenzhen, 518120, China;3.Shandong Shengzhou Marine Biological Technology Co., Ltd. Weihai, 264211, China
Abstract:
The basic nutrients in the enzymatic hydrolysate of sea cucumber were analyzed by national standard methods. The enzymatic hydrolysate further was extracted with organic solvents of different polarities, and the active components were tracked by acetylcholinesterase inhibition self-development. Silica gel column chromatography, Sephadex LH-20 gel column and HPLC were used to obtained components with higher acetylcholinesterase inhibitory activity. In addition, MTT method was used to investigate the effect of total fat-soluble components of sea cucumber enzymatic hydrolysate on the growth of microglial cells (BV-2 cells). LPS-induced cells were used to establish a model of cellular inflammation; and the effects on the levels of NO and IL-6 in LPS-induced BV-2 cells were detected by Griess method and ELISA method. The results showed that the enzymatic hydrolysate contains rich mineral elements like calcium and zinc, as well as high content of unsaturated fatty acids and essential amino acids. From the solvent extract, the TLC spots with acetylcholinesterase inhibitory activity showed no significant UV and fluorescence absorption, and the inhibition rate of the most active component C4-2-E was 91.04% at the concentration of 0.25 μg/μL. The TLF of sea cucumber had no significant inhibitory effect on BV-2 cells within a certain concentration range, and could significantly inhibit the secretion of NO and IL-6 produced by LPS-induced BV-2 cells. Therefore, the TLF of sea cucumber enzymatic hydrolysate may have certain effects of anti-neuroinflammation and Alzheimer's disease, and the chemical structure of specific active substances needs further analysis and identification.
Key words:sea cucumberextraction and separationAlzheimer's diseaseacetylcholinesterase inhibitory activityanti-inflammatory activity





PDF全文下载地址:

点我下载PDF