The Effect of Claudin-5 Overexpression on the Interactions of Claudin-1 and-2 and Barrier Function in Retinal Cells
The Effect of Claudin-5 Overexpression on the Interactions of Claudin-1 and-2 and Barrier Function in Retinal Cells
复制标题
Claudin-5 过表达对 Claudin-1 和 -2 相互作用以及视网膜细胞屏障功能的影响。
DOI：
10.2174/1566524014666141015160355
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发表时间：
2014-01-01
影响因子：
2.5
通讯作者：
Liu, X.
中科院分区：
文献类型：
--
作者：
Tian, R.;Luo, Y.;Liu, X.
Claudin-5是主要的紧密连接蛋白之一，在维持血、脑和视网膜屏障功能方面发挥着重要作用。本研究旨在探讨claudin-5过表达对原代培养的人视网膜色素上皮细胞(HRPECs)和人视网膜内皮细胞(HRECs)中claudin-1和claudin-2的相互作用及屏障功能的影响。慢病毒介导Claudin-5在视网膜细胞中过表达。转染组Claudin-5的mRNA和蛋白水平显著升高。转基因细胞在跨膜上生长3周后，体外建立了稳定的单层细胞屏障模型。Claudins表达分析表明，过表达的claudin-5显著增加了claudin-1的表达，而降低了claudin-2在mRNA和蛋白水平的表达。CO-IP实验和屏障功能分析表明，claudin-5过表达促进了claudin-1和claudin-2的相互作用，增强了视网膜细胞的屏障功能。有趣的是，外源性claudin-5的表达诱导了内源性claudin-5不表达的HRPECs中claudin-5与claudin-1或-2之间的新的相互作用模式。此外，Claudin-5的过表达降低了细胞的迁移率和体外血管形成的萌发能力。本研究表明claudin-5在视网膜屏障的形成中具有正向调节作用。Claudin元件及其相互作用是可以调节的，这种动态特性对于TJ的功能非常重要，从调节视网膜屏障的完整性到连接相关的信号机制。
Claudin-5, one of the dominant tight junctions (TJs) proteins, plays an important role in maintaining the barrier function in the blood brain and retinal barrier. This study aimed to investigate the effect of claudin-5 overexpression on the interactions of claudin-1 and -2 and barrier functions in primary cultured human retinal pigment epithelium cells (HRPECs) and human retina endothelial cells (HRECs). Lentivirus was used to mediate the overexpression of claudin-5 in retinal cells. Significantly increased mRNA and protein levels of claudin-5 were detected in the transfection group. After the transfected cells grew on the transwell membrane for three weeks, a stable monolayer cell barrier model was established in vitro. The claudins expressions analysis showed that overexpressed claudin-5 significantly increased the expression of claudin-1, while it decreased the expression of claudin-2 in both mRNA and protein level. Co-IP experiments and barrier function assay revealed that claudin-5 overexpression promoted the interactions of claudin-1 and claudin-2 and enhanced the barrier function of retinal cells. Intriguingly, the exogenous expression of claudin-5 induced new interaction pattern between claudin-5 and claudin-1 or -2 in HRPECs, which do not have endogenous claudin-5 expression. In addition, claudin-5 overexpression decreased cell mobility and the sprouting capability of vessel tube formation in vitro. This study demonstrated that claudin-5 has a positive regulation in the formation of retinal barrier. Claudin elements and their interactions can be modulated and that such dynamic properties are important for the functions of TJs, ranging from the regulation of retinal barrier integrity to junction-associated signaling mechanisms.