果糖偶联壳聚糖两亲性递送体系的构建及其诱导GLUT2调控的胞吞作用机制研究

食源性亲水与疏水活性物质具有良好的生理活性且存在广泛的协同增效现象，但由于较低的小肠吸收效率，经口摄入后难以在体内有效发挥生物活性。果糖修饰型递送体系可通过葡萄糖转运体2（GLUT2）调控的胞吞作用显著提高活性物质的小肠吸收效率，但胞吞作用机制尚不明确。本项目拟以果糖偶联壳聚糖和β-环糊精为基础，通过层层自组装法构建共载蛋清肽（亲水）和姜黄素（疏水）的两亲性递送体系；采用体内和体外吸收模型，阐明该递送体系结构与蛋清肽和姜黄素跨膜共转运效率的关系；综合运用免疫荧光共定位、生物素标记及蛋白质组学等技术，探究果糖偶联壳聚糖两亲性递送体系胞吞作用过程中对GLUT2位置迁移、受体功能转变、磷酸化修饰、表达量及表达方向等方面的影响，从分子和细胞水平上阐明果糖偶联壳聚糖两亲性递送体系诱导GLUT2调控的胞吞作用机制。本项目将为食源性亲水与疏水活性物质在食品产业中的研究与应用提供重要的科学依据和理论基础。

Food-derived hydrophilic and hydrophobic bioactive compounds show excellent bioactivity and extensive synergistic effects. However, due to the poor intestinal permeability, the bioactivity of multi-components is greatly limited after oral administration. Fructose modified delivery system can significantly improve the intestinal permeability of bioactive compounds through endocytosis mediated by glucose transporter 2 (GLUT2), while its transport mechanism is not yet clear. Based on the fructose-conjugated chitosan and β-cyclodextrin, an amphiphilic delivery system co-loaded with egg white peptide (hydrophilicity) and curcumin (hydrophobicity) will be fabricated by the layer-by-layer self-assembly. The fructose-conjugated chitosan amphiphilic delivery system will be performed to clarify the relationship between the structure of the delivery system and the transmembrane transport efficiency of egg white peptide and curcumin by in vivo and in vitro intestinal absorption models. Furthermore, immunofluorescence co-localization, biotin labeling, and proteomics will be used to explore the endocytosis mechanism of the GLUT2-mediated fructose-conjugated chitosan amphiphilic delivery system. Moreover, the effects of fructose-conjugated chitosan amphiphilic delivery system on the site migration, receptor function transition, phosphorylation, and expression quantity and direction of GLUT2 will be considered as the main factors to evaluate the endocytosis mechanism regulated by GLUT2 at the molecular and cellular level. This project will provide critical scientific evidence and a theoretical foundation for applying of food-derived hydrophilic and hydrophobic bioactive compounds in the food industry.