海参岩藻聚糖硫酸酯的空间构象解析及干预胰岛素抵抗的定量构效关系研究

项目名称： 海参岩藻聚糖硫酸酯的空间构象解析及干预胰岛素抵抗的定量构效关系研究

项目编号： No.31471684

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 常耀光

作者单位： 中国海洋大学

项目金额： 83万元

中文摘要： 项目拟针对海参岩藻聚糖硫酸酯（SC-FUC）干预胰岛素抵抗的构效关系展开深入研究。以多种低值海参为原料提取SC-FUC，利用自主筛选的糖苷酶特色制备系列酶解产物，基于多级质谱解明其精细一级结构；采用高效凝胶排阻色谱-多角度激光光散射、动态光散射、原子力显微镜等技术解析SC-FUC及其酶解产物的空间构象，进而阐明一级结构对SC-FUC空间构象的影响；运用胰岛素抵抗细胞模型及动物模型，研究SC-FUC及其酶解产物对胰岛素抵抗的干预作用，并以PI3K/PKB通路中关键靶点基因的表达为切入点，探明作用的分子机制；在此基础上，基于人工神经网络建立定量构效关系模型，准确阐明SC-FUC干预胰岛素抵抗作用与其硫酸酯化形式、分子量、链构象及链尺寸间的构效关系。本项目将首次从多个结构水平阐明SC-FUC的分子结构，并建立首个SC-FUC定量构效关系模型，为SC-FUC深度开发奠定理论基础，推动海参产业发展。

中文关键词： 海参；岩藻聚糖硫酸酯；空间构象；胰岛素抵抗；定量构效关系

英文摘要： This project is set to investigate the structure-activity relationship of insulin resistance-affecting function of sea cucumber fucoidan (SC-FUC). The program is as follows. Low-price sea cucumbers are employed as materials to extract SC-FUC, and by using a special self-isolated fucoidanase, a series of SC-FUC enzymatic degradation products would be prepared; the delicate primary structure of SC-FUC would be clarified by using ESI-MS/MS; the conformation of SC-FUC and its degrdation products would be studied by using a combination of HPLC-multiple angle laser light scattering, dynamic light scattering and atomic force microscopy, and the relationship between primary structure and confromation would be further elucidated; insulin resistance-affecting activity of SC-FUC and its degradation product would be investigated by using both of the cell model and animal model, and the molecualr mechanism would be clarified via phosphatidylinositol 3-hydroxy kinase and Protein Kinase B (PI3K/PKB) pathway; finally, the QSAR model would be established based on artificial neural network, and in that case, the relationship between the insulin resistance-affecting activity and several structural features including sulfatation pattern, molecular mass, chain morphology, and chain dimension could be clearly elucidated. This study would clarify the molecular structure of SC-FUC in different structural levels at the first time, and establish the first quantitative structure-activity ralationship (QSAR) model of SC-FUC. It would provide academic support to the efficient utilization of SC-FUC, and therefore be benefit to the benign development of sea cucumber industry.

英文关键词： sea cucumber;fucoidan;conformation;insulin resistance;quantitative structure-activity relationship