基于Caco-2细胞的外源核酸吸收模型的建立及其吸收机制.pdfVIP

236 201 6, Vo1.37, No.23 画面副事 ※营养E生 基于Caco-2细胞的外源核酸吸收模型的建主及其 吸收机制 王晓倩，张艳芳，王倩，高晓梦，李春}I I.李敬，吴薇，董平气梁兴国 (中国海洋大学食品科学与工程学院，山东青岛 266∞3) 摘 要z 基于人结肠癌细胞系Caco-2细胞建立外源核酸的吸收模型，并在此模型的基础上探究质粒DNA的肠道吸 收机制。在Transwell孔膜上培养Caco-2细胞21 d后，通过透射电子显微镜观察细胞形态、监测培养期间跨膜电阻 (transepithelial electrical resistance , TEER) 值、测定荧光黄透过率评价细胞模型是否成功建成:通过细胞毒性实 验探究质粒对细胞生长的影响:将质粒DNA加入细胞模型中进行双向转运实验，探究质粒的肠道吸收规律:在低 温和添加特异性抑制剂条件下进行质粒转运实验，进一步推测质粒的肠道吸收机制。结果表明，细胞分化形态良 好、 TEER值及荧光黄表现透过系数 (apparent permeability coefficient , P叩p) 均符合要求，细胞模型可用于转运实 验:质粒对细胞生长无显著抑制作用，可用于转运实验:随着时间的延长，质粒在模型两方向上的转运均呈逐渐饱 和趋势， P app (肠腔侧 (apical. AP )→基底侧 (basolateral ， BL) )远大于P啊 (BL→AP) ，提示质粒的转运受到 某种载体蛋白的介导，膜介导的跨细胞运输方式可能参与其中:在低温和添加特异性抑制剂条件下，质粒的转运均 受到显著抑制，进一步表明读过程是一个跨细胞方式的主动运输过程。 关键词: Caco-2细胞模型:外源核酸:吸收机制:质粒DNA Absorption Model and Mechanism of Exogenous Nucleic Acids Based on Caco-2 Cells WANG Xia叫ian， ZHANG Yanfang , WANG Qian, GAO Xiaomeng , LI Chunchu钮， LI Ji吨， WU Wei , DONG Ping* , LIANG Xingguo (College of Food Science and Engineering, Ocean University of China, Qingdao 266∞3， China) Abstract: In this study, we employed Caco-2 cells ωestablish an absorption model of exogenous nucleic acids. Using 也is model , we also explored the mechanism of intestinal absorption of plasmid DNA. After culturing Caco-2 cells for 21 days on Transwell membranes , cell morphology, transepithelial electrical resistance (TEER) and apparent permeability coeffi.cient (P，啊) offiuorescent yellow were measured and used to evaluate whether the model was established su∞essfully. C}仙toxicity experiments were used to explore the e能ct of plasmid DNA on cell gro啊th. The mechanism of intestinal DNA absorption was explored by two-way transfer experiments. To