Oxidative stress-induced activation of Abl and Src kinases rapidly induces P-glycoprotein internalization via phosphorylation of caveolin-1 on tyrosine-14, decreasing cortisol efflux at the blood-brain barrier
Oxidative stress-induced activation of Abl and Src kinases rapidly induces P-glycoprotein internalization via phosphorylation of caveolin-1 on tyrosine-14, decreasing cortisol efflux at the blood-brain barrier
复制标题
DOI：
10.1177/0271678x18822801
复制
发表时间：
2020-02-01
影响因子：
6.3
通讯作者：
Terasaki, Tetsuya
中科院分区：
文献类型：
--
作者：
Hoshi, Yutaro;Uchida, Yasuo;Terasaki, Tetsuya
脑缺血再灌注期间暴露于高水平的糖皮质激素诱导神经元细胞死亡。在缺血再灌注过程中，氧化应激改变血脑屏障（BBB）的功能，因此我们假设它可能会损害P-糖蛋白（P-gp）介导的糖皮质激素在BBB的外排转运。因此，本研究的目的是阐明这种假定的P-gp介导的外排功能降低的分子机制。首先，我们建立了H2 O2治疗的人在体外血脑屏障模型（hCMEC/D3）减少P-gp外排转运活性和蛋白表达的质膜上在20分钟内。这些结果表明，外排功能的快速下降可能是由于P-gp的内化。此外，过氧化氢处理显着增加酪氨酸-14-磷酸化小窝蛋白-1，这是参与P-gp的内化。大鼠脑灌注研究表明，皮质醇流出在血脑屏障显着减少过氧化氢管理，和抑制剂的Abl激酶和Src激酶，磷酸化酪氨酸-14小窝蛋白-1，抑制这种减少。总体而言，这些研究结果支持的想法，氧化应激诱导的激活Abl激酶和Src激酶诱导P-gp的内化通过磷酸化的酪氨酸-14小窝蛋白-1，导致快速减少P-gp介导的皮质醇流出在血脑屏障。
Exposure of the brain to high levels of glucocorticoids during ischemia-reperfusion induces neuronal cell death. Oxidative stress alters blood-brain barrier (BBB) function during ischemia-reperfusion, and so we hypothesized that it might impair P-glycoprotein (P-gp)-mediated efflux transport of glucocorticoids at the BBB. Therefore, the purpose of this study was to clarify the molecular mechanism of this putative decrease of P-gp-mediated efflux function. First, we established that H2O2 treatment of a human in vitro BBB model (hCMEC/D3) reduced both P-gp efflux transport activity and protein expression on the plasma membrane within 20 min. These results suggested that the rapid decrease of efflux function might be due to internalization of P-gp. Furthermore, H2O2 treatment markedly increased tyrosine-14-phosphorylated caveolin-1, which is involved in P-gp internalization. A brain perfusion study in rats showed that cortisol efflux at the BBB was markedly decreased by H2O2 administration, and inhibitors of Abl kinase and Src kinase, which phosphorylate tyrosine-14 in caveolin-1, suppressed this decrease. Overall, these findings support the idea that oxidative stress-induced activation of Abl kinase and Src kinase induces internalization of P-gp via the phosphorylation of tyrosine-14 in caveolin-1, leading to a rapid decrease of P-gp-mediated cortisol efflux at the BBB.