糙海参极性脂的提取和神经保护活性  

Extraction and Neuroprotective Activity of Polar Lipid from Holothuria scabra

作　　者：熊仕洪 张翼 赵佳文 周龙建 刘亚月 杨志友 张永平 XIONG Shihong;ZHANG Yi;ZHAO Jiawen;ZHOU Longjian;LIU Yayue;YANG Zhiyou;ZHANG Yongping(College of Food Science and Technology,Guangdong Ocean University/Guangdong Provincial Key Laboratory of Aquatic Product Processing and Safety/Guangdong Provincial Engineering Laboratory for Marine Biological Products/Guangdong Provincial Engineering Technology Research Center of Seafood/Key Laboratory of Advanced Processing of Aquatic Product of Guangdong Higher Education Institution/Guangdong Provincial Center for Modern Agricultural Scientific Innovation/Marine Biomedicine Research and Development Center at Shenzhen Institute of Guangdong Ocean University/Zhanjiang Municipal Key Laboratory of Marine Drugs and Nutrition for Brain Health/Research Institute for Marine Drugs and Nutrition,Guangdong Ocean University,Zhanjiang 524088,China;Southern Marine Science and Engineering Guangdong Laboratory(Zhanjiang),Zhanjiang 524006,China;Collaborative Innovation Center of Seafood Deep Processing,Dalian Polytechnic University,Dalian 116034,China)

机构地区：[1]广东海洋大学食品科技学院/广东省水产品加工与安全重点实验室/广东省海洋生物制品工程实验室/广东省海洋食品工程技术研究中心/水产品深加工广东普通高等学校重点实验室/广东省现代农业科技创新中心/广东海洋大学深圳研究院海洋医药研发中心/湛江市脑健康海洋药物与营养品重点实验室/广东海洋大学海洋药物研究所,广东湛江524088 [2]南方海洋科学与工程广东省实验室(湛江),广东湛江524006 [3]海洋食品精深加工关键技术省部共建协同创新中心/大连工业大学,辽宁大连116034

出　　处：《广东海洋大学学报》2025年第1期94-105,共12页Journal of Guangdong Ocean University

基　　金：广东省普通高校重点领域专项(生物医药与健康)(2021ZDZX2064);广东省基础与应用基础研究基金自然科学基金(面上项目)(2022A1515010783);深圳市科创委基础研究面上项目(JCYJ20220530162014032);广东省科技专项资金-基础与应用基础研究专题(2021A05240);湛江市海洋青年人才创新项目(2022E05010)。

摘　　要：【目的】研究糙海参(Holothuria scabra)极性脂的抗阿尔茨海默氏症作用。【方法】采用8种总脂提取方法结合冷丙酮抽提制备海参极性脂,计算不同方法下的极性脂得率,并通过高效液相色谱分析比较所制备极性脂的指纹图谱,采用薄层层析-磷钼酸显色法和基于液质联用的脂质组注释分析其成分,对通过优选提取方法得到的总极性脂,采用DPPH自由基清除法、乙酰胆碱酯酶抑制活性测试、LPS诱导的小胶质细胞(BV-2)神经炎症模型、Aβ_(25-35)诱导的小鼠海马神经元(HT-22)损伤模型等指标,综合评价糙海参极性脂的抗阿尔茨海默氏症潜力。【结果】1)在8种不同的总脂提取方法中,Bligh&Dyer法既有较高的最终极性脂得率(1.25%),且提取的极性脂成分多样,并在同等效果下氯仿消耗量最低,该方法在极性脂提取上综合表现最佳;2)薄层层析-磷钼酸显色实验显示该极性脂中含有磷脂类成分,脂质组分析显示其含有丰富的磷脂、鞘脂、甘油脂、脂肪酸、N-磺甲基酰胺和未知脂类等成分;3)糙海参极性脂表现出较好的DPPH自由基清除活性[半数清除质量浓度(217.13±2.34)μg/mL]、乙酰胆碱酯酶抑制活性[半抑制质量浓度(30.39±1.48)μg/mL]、抑制LPS诱导的BV-2细胞中NO生成活性(1μg/mL剂量下抑制率为48.88%),以及对Aβ_(25-35)诱导的HT-22神经元损伤的治疗性、预防性保护作用和清除胞内活性氧的作用(40μg/mL剂量下)。【结论】糙海参极性脂具有较好的多靶向抗阿尔茨海默氏症体外活性,作为相关功能食品和药物具备进一步研究开发的价值。【Objective】To study the Anti-AD(Alzheimer’s disease)effect of polar lipids(PLs)from Holothuria scabra.【Methods】The total H.scabra PL(HSPL)was prepared by use of eight total lipid extraction methods followed by cold acetone extraction.The different extraction techniques were compared for their PL yields and phospholipid fingerprints by high-performance liquid chromatography(HPLC).The composition of the HSPL was primarily analyzed by thin-layer chromatography(TLC)with phosphomolybdic acid staining and lipidomics annotation based on liquid chromatography coupled with mass spectrometry(LC-MS).The total HSPL prepared by the optimized method was comprehensively evaluated for its anti-AD potential using different bioassays,including 2,2-diphenyl-1-picrylhydrazyl(DPPH)free radical scavenging activity,acetylcholinesterase(AChE)inhibitory activity,lipopolysaccharide(LPS)-induced neuroinflammatory model in microglial cell(BV-2),and Aβ_(25-35)-induced neuronal injury model in mouse hippocampus cell(HT-22).【Result】1)Among the eight lipid extraction methods,the Bligh&Dyer method was the most effective for polar lipid preparation,with the highest polar lipid yield(1.25%),the most diverse lipid composition,and the lowest chloroform use.2)TLC-phosphomolybdic acid staining displayed the presence of phospholipid in the HSPL and lipidomics analysis revealed rich phospholipids,sphingolipids,glycerolipids,fatty acids,N-sulfomethylamides,and unannotated lipids as its possible ingredients.3)The HSPL demonstrated favorable DPPH free radical scavenging activity with a semi-scavenging mass concentration of(217.13±2.34)μg/mL,AChE inhibitory activity with a semi-inhibitory mass concentration of(30.39±1.48)μg/mL,and inhibition to LPS-induced NO production in BV-2 cells with an inhibitory rate of 48.88%at the dose of 1μg/mL.Besides,these lipids exhibited both therapeutic and preventive protective effects against Aβ_(25-35)-induced neuronal damage in HT-22 cells,as well as the ability to scavenge intracellular reactive oxyg

关 键 词：糙海参 极性脂 提取 阿尔茨海默氏症 神经保护 

分 类 号：O629.6[理学—有机化学]