Inhibition of glycogen synthase kinase 3β promotes tight junction stability in brain endothelial cells by half-life extension of occludin and claudin-5.
Inhibition of glycogen synthase kinase 3β promotes tight junction stability in brain endothelial cells by half-life extension of occludin and claudin-5.
复制标题
DOI：
10.1371/journal.pone.0055972
复制
发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Persidsky Y
中科院分区：
文献类型：
--
作者：
Ramirez SH;Fan S;Dykstra H;Rom S;Mercer A;Reichenbach NL;Gofman L;Persidsky Y
神经炎症性疾病通常涉及血脑屏障 (BBB) 功能障碍。因此，识别能够维持屏障保真度的分子靶标具有临床重要性。我们之前曾报道过糖原合成酶激酶 3β (GSK3β) 抑制对原代人脑内皮细胞的抗炎作用。在这里，我们表明 GSK3β 抑制剂还通过影响紧密连接 (TJ) 蛋白的稳定性来促进屏障的紧密性。跨内皮电阻 (TEER) 用于评估药物抑制剂和 GSK3β 突变体的屏障完整性。 GSK3β 的抑制使 TEER 逐渐持续增加（比基线高出 22%）。对亚细胞膜组分的分析显示，必需的紧密连接蛋白、occludin 和 claudin-5 的数量有所增加，但 claudin-3 的数量没有增加。这种现象归因于 TJ 蛋白更新的减少，而不是转录调控。使用一种新的基于细胞的测定法，GSK3β 的失活使 occludin 和 claudin-5 的半衰期分别显着延长了 32% 和 43%。作为 GSK3β 抑制的函数，β-连环蛋白与 ZO-1 的关联增强之间也建立了相关性。总的来说，我们的研究结果表明，可以使用 GSK3β 抑制剂作为延长关键紧密连接蛋白半衰期的手段，以促进神经炎症期间 BBB 的重新密封。
Neuroinflammatory conditions often involve dysfunction of the Blood-Brain Barrier (BBB). Therefore, identifying molecular targets that can maintain barrier fidelity is of clinical importance. We have previously reported on the anti-inflammatory effects that glycogen synthase kinase 3β (GSK3β) inhibition has on primary human brain endothelial cells. Here we show that GSK3β inhibitors also promote barrier tightness by affecting tight junction (TJ) protein stability. Transendothelial electrical resistance (TEER) was used to evaluate barrier integrity with both pharmacological inhibitors and mutants of GSK3β. Inhibition of GSK3β produced a gradual and sustained increase in TEER (as much as 22% over baseline). Analysis of subcellular membrane fractions revealed an increase in the amount of essential tight junction proteins, occludin and claudin-5, but not claudin-3. This phenomenon was attributed to a decrease in TJ protein turnover and not transcriptional regulation. Using a novel cell-based assay, inactivation of GSK3β significantly increased the half-life of occludin and claudin-5 by 32% and 43%, respectively. A correlation was also established between the enhanced association of β-catenin with ZO-1 as a function of GSK3β inhibition. Collectively, our findings suggest the possibility of using GSK3β inhibitors as a means to extend the half-life of key tight junction proteins to promote re-sealing of the BBB during neuroinflammation.
登录
查看更多内容
DOI：
10.1152/ajprenal.00674.2009
发表时间：
2010-08-01
影响因子：
4.2
作者：
Raikwar, Nandita S.;Vandewalle, Alain;Thomas, Christie P.
通讯作者：
Thomas, Christie P.
影响因子：
3.3
作者：
Medici, D;Hay, ED;Goodenough, DA
通讯作者：
Goodenough, DA
Wnt/β-catenin信号传导控制血脑屏障的发展。
影响因子：
7.8
作者：
Liebner, Stefan;Corada, Monica;Bangsow, Thorsten;Babbage, Jane;Taddei, Andrea;Czupalla, Cathrin J.;Reis, Marco;Felici, Angelina;Wolburg, Hartwig;Fruttiger, Marcus;Taketo, Makoto M.;von Melchner, Harald;Plate, Karl Heinz;Gerhardt, Holger;Dejana, Elisabetta
通讯作者：
Dejana, Elisabetta
DOI：
10.1083/jcb.132.3.451
发表时间：
1996-02
期刊：
The Journal of cell biology
影响因子：
--
作者：
Rajasekaran AK;Hojo M;Huima T;Rodriguez-Boulan E
通讯作者：
Rodriguez-Boulan E
影响因子：
4.8
作者：
Liu, F;Schaphorst, KL;Garcia, JGN
通讯作者：
Garcia, JGN