小肠上皮细胞OATP1A2和OATP2B1载体介导的蛋清源活性寡肽完整吸收机制

食源性生物活性寡肽在小肠上皮的完整吸收机制是活性肽研究领域亟待解决的关键问题，制约着活性肽类营养食品的研究和发展。本课题组前期研究表明有机阴离子转运多肽载体OATP1A2和OATP2B1可能参与寡肽在小肠上皮的完整吸收。本项目以蛋清源活性寡肽为对象，构建高表达OATP1A2/OATP2B1细胞模型，利用HPLC-MS、RT-PCR和PRM技术研究蛋清源活性寡肽的完整吸收与OATP1A2/OATP2B1表达之间的关系，利用荧光标记与共聚焦激光显微镜技术研究蛋清源活性寡肽的完整吸收过程，从基因、蛋白和细胞水平确证小肠上皮细胞OATP1A2和OATP2B1对蛋清源活性寡肽的完整转运作用；利用动力学方法研究寡肽完整吸收的结构选择性、离子依赖性和细胞膜电位变化等特性，阐明小肠上皮细胞OATP1A2和OATP2B1介导的蛋清源活性寡肽完整吸收机制。本项目将为蛋清源活性寡肽在小肠上皮的完整吸收提供依据。

The transport mechanisms of food-derived bioactive oligopeptides through the intestinal epithelium in intact forms becomes to be a critical issue that influencing the development of the industry of nutritional food based on food-derived bioactive peptides. Our previous study reveals that organic anion transporting polypeptides OATP1A2 and OATP2B1 may take responsibility for the transport of oligopeptides in intact forms through the intestinal epithelium. The objective of this project is to investigate the OATP1A2 and OATP2B1 mediated transport of egg white-derived bioactive oligopeptides in the model with overexpressed of OATP1A2 and OATP2B1. HPLC/HPLC-MS, RT-PCR and PRM will be used to study the relationship between the intact transport of egg white-derived bioactive oligopeptides and the transcription and expression of OATP1A2 and OATP2B1. Fluorescence labeling and confocal laser microscopy will be used to trace the transport progress of egg white-derived bioactive oligopeptides. It helps to confirm the critical role of OATP1A2 and OATP2B1 in the absorption of oligopeptides in the intestinal epithelium on gene, protein and cell levels. According to the method of transport kinetics, the transport characteristic of oligopeptides, including substrate specificity, H+ or Na+ dependence and cell membrane potential changes will be charactered. Those will help to figure out and explain the transport mechanisms of oligopeptides in the intestinal epithelium mediated by OATP1A2 and OATP2B1. In a word, this work will provide basis for the intact transport of egg white-derived bioactive peptides in the intestinal epithelium mediated.