基于高分辨质谱及斑马鱼模型的磷虾油磷脂组成特征及降血糖活性分析  

Composition Characteristics and Hypoglycaemic Activity of Phospholipids in Krill Oil as Determined by High-Resolution Mass Spectrometry and Zebrafish

作　　者：刘慧茹 李文玉 陈立国 王昊 周芳 韩利文 LIU Huiru;LI Wenyu;CHEN Liguo;WANG Hao;ZHOU Fang;HAN Liwen(The School of Pharmacy,Shandong First Medical University&Shandong Academy of Medical Sciences,Jinan 250117,China;Fengshi(Qingdao)Marine Technology Co.Ltd.,Qingdao 266100,China)

机构地区：[1]山东第一医科大学(山东省医学科学院)药学院(药物研究所),山东济南250117 [2]逢时(青岛)海洋科技有限公司,山东青岛266100

出　　处：《食品科学》2024年第18期134-142,共9页Food Science

基　　金：山东第一医科大学学术提升计划项目(2019LJ003);山东省中医药高层次人才培育项目。

摘　　要：为了探究磷虾油的磷脂成分组成特征及降血糖活性,本研究首先采用比色法测定磷虾油中总磷脂含量,采用高分辨质谱对磷虾油中的磷脂组成成分进行检测并运用质谱裂解规律以及Lipidmaps、Lipidblast等数据库进行结构解析;此外采用四氧嘧啶与蔗糖联合处理构建斑马鱼高血糖模型,检测磷虾油样品对斑马鱼血糖及与血糖调节相关的生化指标和基因表达水平;进一步通过分子对接技术探究磷虾油特有磷脂型二十二碳六烯酸/二十碳五烯酸(docosahexaenoic acid/eicosapentaenoic acid,DHA/EPA)降血糖潜在作用位点。结果显示,冷萃磷虾油中磷脂含量可达57.57%,使用高分辨质谱技术共鉴定出63种磷脂,其中磷脂酰胆碱种类最丰富;磷虾油可显著降低高血糖斑马鱼体内的血糖浓度,升高胰岛素、糖原水平,抑制FOXO1/PCK1/G6PC的表达,促进GLUT4的表达;通过分子对接发现,磷虾油特有磷脂型DHA/EPA可能作用在FOXO1/PCK1/G6PC位点,提示磷脂成分与降血糖活性密切相关。综上,本研究发现冷萃工艺制备的磷虾油富含磷脂类成分,具有较好的降血糖效果,可作为一种潜在的降血糖功能天然产物。In this study,in order to investigate the phospholipid composition characteristics and hypoglycemic activity of krill oil,we determined the content of total phospholipids in krill oil by a colorimetric method,examined the phospholipid composition by high-resolution mass spectrometry,and then applied the mass spectral fragmentation pattern and the databases Lipidmaps and Lipidblast for structural analysis.Furthermore,we evaluated the effect of krill oil on blood glucose,biochemical indexes and gene expression levels related to the regulation of blood glucose in a zebrafish model of hyperglycemia induced by the combined treatment of alloxan and sucrose,and explored the potential sites of hypoglycemic action of krill oil-specific phospholipid docosahexaenoic acid(DHA)and eicosapentaenoic acid(EPA)by molecular docking.The results showed that the content of phospholipids in cold-extracted krill oil was 57.57%,and a total of 63 phospholipids were identified,of which phosphatidylcholine(PC)was the most abundant.Krill oil significantly reduced blood glucose concentration,elevated insulin and glycogen levels,inhibited the expression of FOXO1/PCK1/G6PC,and promoted the expression of GLUT4 in hyperglycemic zebrafish.Molecular docking revealed that DHA and EPA in krill oil might act at the FOXO1/PCK1/G6PC sites,suggesting close relationship between the phospholipid components and the hypoglycemic activity of krill oil.In summary,this study found that cold-extracted krill oil is rich in phospholipids and is a potential natural product with good hypoglycemic effect.

关 键 词：磷虾油 磷脂 高分辨质谱 降血糖 斑马鱼模型 

分 类 号：R151[医药卫生—营养与食品卫生学]