柑橘黄酮类化合物的肠吸收构-效关系及关键外排蛋白作用机制研究

项目名称： 柑橘黄酮类化合物的肠吸收构-效关系及关键外排蛋白作用机制研究

项目编号： No.31471625

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 食品科学、农学基础与作物学

项目作者： 徐晓云

作者单位： 华中农业大学

项目金额： 84万元

中文摘要： 黄酮类化合物在植物界中大量存在并具多种活性，然而其口服后生物利用度较低严重制约其进一步应用。本课题以30种主要来源于柑橘的黄酮类化合物为研究对象，从肠道吸收转运角度，将其作用于Caco-2细胞，构建结构-渗透3DQSPR模型，比较不同结构黄酮类化合物肠道吸收的结构特征。为明确其肠吸收的限制因素，进一步研究两个关键外排蛋白P-gp和MRP1的作用机制，利用外排蛋白P-gp或MRP1过表达的KB细胞，将黄酮类化合物作用于外排蛋白介导的道诺霉素转运，构建两个结构-亲和力3D QSAfR模型，与同源模建得到的人P-gp或MRP1分别进行分子对接，寻找黄酮类化合物的蛋白结合位点及方式，和QSAfR结果对比、验证，获得黄酮类化合物与外排蛋白结合的必要结构。结合肠道吸收实验，分析肠道中易吸收的黄酮类化合物的关键结构位点。用在体肠道和体外肠道模型验证，为筛选或合成具有高吸收结构特征的黄酮类化合物提供依据。

中文关键词： 黄酮类化合物；人结肠腺癌细胞；定量构-效关系；分子对接

英文摘要： Flavonoids are ubiquitously compounds, presenting in the plant world and our common diet. Although epidemiological investigations and laboratory studies have indicated their pharmacological properties, it is a big challenge for flavonoids further development due to their low oral bioavailability. In this study, 30 kinds of flavonoids will be studied in vitro intestinal model Caco-2 monolayers, getting a quantitative structure permeability relationship (QSPR) to define the structural requirements of flavonoids necessary for high Papp. In order to find out limiting factor for low bioavailability and help on understanding the mechanism of the P-gp and MRP1, the affinity of flavonoids to this two crucial transporters will be studied in P-gp overexpression or MRP1 transfected KB cells. The results will be used to define the necessary structural requirements of flavonoids for affinity of P-gp or MRP1-mediated daunorubicin transport in the cellular models, and two quantitative structure affinity relationships (QSAfR) model will be obtained and analyzed. Flavonoids will docking with P-gp or MRP1 obtained by homology modeling to find their protein binding sites and binding mode. The expected results will compared with the data from QSAfR to get the essential structure combined with transporters. Taking the results of QSPR into account, the structure of flavonoids easy to be absorbed in intestine will be figured out and comfirmed by vivo intestinal and vitro intestinal model. The crucial structure information will be a useful reference for screening of high absorbtion rate flavonoids or its structural modification.

英文关键词： Flavonoids;Caco-2 Cells;QSAR;Molecular Docking