Takizawa Yusuke, Kishimoto Hisanao, Tomita Mikio, Hayashi Masahiro
Department of Drug Absorption and Pharmacokinetics, School of Pharmacy and Life Sciences, Tokyo University of Pharmacy and Life Sciences, 1432-1, Horinouchi, Hachioji, Tokyo, 192-0392, Japan,
Eur J Drug Metab Pharmacokinet. 2014 Sep;39(3):211-20. doi: 10.1007/s13318-013-0151-z. Epub 2013 Sep 8.
Tight junction (TJ) is composed of the most apical components of the intercellular junctional complex in epithelial cells; TJ has cell polarity and functions as a major determinant of epithelial barrier function. In this study, to clarify the components of TJ required for its reconstruction and functional acquisition, we examined the changes in intestinal mucosal structure that depended on mucosal lesion by intestinal I/R, that is, the changes in mRNA and protein expression of the claudin family and scaffold proteins. We used an in vivo intestinal I/R model made using the spring scale and surgical sutures, and examined the mRNA and protein expression levels of TJ components by real-time RT-PCR and Western blotting, respectively. Changes in mRNA and protein expression levels of TJ components by intestinal I/R were observed. Among them, characteristic changes were observed in claudin-2 and claudin-4. In addition, the expression behavior of multi-PDZ domain protein (MPDP) mRNA was similar to that of claudin-4. In conclusion, in the recovery process of TJ from mucosal lesion by intestinal I/R, it was suggested that claudin-2 and claudin-4 strongly participate in the reconstruction and functional acquisition of TJ, respectively. Furthermore, it was suggested that MPDZ, which is scaffold protein, also has an important role in these processes.
紧密连接(TJ)由上皮细胞间连接复合体最顶端的成分组成;TJ具有细胞极性,是上皮屏障功能的主要决定因素。在本研究中,为了阐明TJ重建和功能获得所需的成分,我们研究了肠道缺血/再灌注(I/R)引起的肠道黏膜损伤所导致的肠道黏膜结构变化,即紧密连接蛋白家族和支架蛋白的mRNA及蛋白表达变化。我们使用弹簧秤和手术缝线制作了体内肠道I/R模型,并分别通过实时逆转录聚合酶链反应(RT-PCR)和蛋白质免疫印迹法检测TJ成分的mRNA和蛋白表达水平。观察到肠道I/R引起的TJ成分mRNA和蛋白表达水平变化。其中,紧密连接蛋白-2(claudin-2)和紧密连接蛋白-4(claudin-4)出现了特征性变化。此外,多PDZ结构域蛋白(MPDP)mRNA的表达行为与claudin-4相似。总之,在肠道I/R导致黏膜损伤后TJ的恢复过程中,提示claudin-2和claudin-4分别强烈参与TJ的重建和功能获得。此外,提示作为支架蛋白的MPDZ在这些过程中也具有重要作用。