TIMP-1 通过 MMP 保护脑内皮细胞紧密连接免受降解。
TIMP-1 Protects Tight Junctions of Brain Endothelial Cells From MMP-Mediated Degradation.
机构信息
Department of Chemical and Materials Engineering, University of Nevada, 1664 N. Virginia St, Reno, NV, 89557, USA.
Department of Pharmacology, University of Nevada, Reno School of Medicine, Reno, NV, USA.
出版信息
Pharm Res. 2023 Sep;40(9):2121-2131. doi: 10.1007/s11095-023-03593-y. Epub 2023 Sep 12.
OBJECTIVE
The blood-brain barrier (BBB) plays a critical role in central nervous system homeostasis, and the integrity of BBB is disrupted in many neurodegenerative diseases. Matrix metalloproteinases (MMPs) degrade the tight junctions (TJs) of endothelial cells and basement membrane components essential to BBB integrity, which leads to increased BBB permeability and allows inflammatory cells and neurotoxic substances to enter the brain. Tissue inhibitors of metalloproteinases (TIMPs), endogenous inhibitors of MMPs, regulate MMP activity, thereby maintaining BBB integrity.
METHODS
The disruptive impacts of MMP-3 and MMP-9 on BBB and protective effect of TIMP-1 were investigated in a simplified in vitro model of the BBB, which was generated using rat brain microvascular endothelial cells (RBMEC). The main features of BBB formation, including permeability and the trans-endothelial electrical resistance (TEER), were monitored over time after the addition of MMP-3 and MMP-9 and their complexes with TIMP-1 inhibitor.
RESULTS
Our results indicated that MMP-3 and MMP-9 caused a dose-dependent disruption of the BBB, with 1.5 µM MMPs resulting in an over threefold increase in permeability, while TIMP-1 inhibition protected the integrity of the BBB model and recovered TEER and permeability of RBMECs. The disruption and recovery of tight junction proteins of RBMECs after MMP and TIMP treatment were also detected using fluorescent microscopy.
CONCLUSION
MMP-9 and MMP-3 disrupt the BBB by degrading tight junctions in endothelial cells, and TIMP-1 could inhibit the disruptive effect of MMP-3 and MMP-9 by showing potential as therapeutic protein against MMP-related diseases where BBB disruption plays a role.
目的
血脑屏障（BBB）在中枢神经系统稳态中起着关键作用，而许多神经退行性疾病会破坏 BBB 的完整性。基质金属蛋白酶（MMPs）降解内皮细胞紧密连接（TJ）和基底膜成分，这些成分对 BBB 的完整性至关重要，导致 BBB 通透性增加，使炎症细胞和神经毒性物质进入大脑。基质金属蛋白酶抑制剂（TIMPs）是 MMPs 的内源性抑制剂，可调节 MMP 活性，从而维持 BBB 的完整性。
方法
在 BBB 的简化体外模型中研究了 MMP-3 和 MMP-9 对 BBB 的破坏作用以及 TIMP-1 的保护作用，该模型是使用大鼠脑微血管内皮细胞（RBMEC）产生的。在添加 MMP-3 和 MMP-9 及其与 TIMP-1 抑制剂的复合物后，随时间监测 BBB 形成的主要特征，包括通透性和跨内皮电阻（TEER）。
结果
结果表明，MMP-3 和 MMP-9 导致 BBB 呈剂量依赖性破坏，1.5µM MMPs 可使通透性增加三倍以上，而 TIMP-1 抑制可保护 BBB 模型的完整性，并恢复 RBMEC 的 TEER 和通透性。还使用荧光显微镜检测了 MMP 和 TIMP 处理后 RBMEC 紧密连接蛋白的破坏和恢复情况。
结论
MMP-9 和 MMP-3 通过降解内皮细胞中的紧密连接破坏 BBB，而 TIMP-1 可能通过抑制 MMP-3 和 MMP-9 的破坏作用显示出作为治疗 MMP 相关疾病的潜在治疗蛋白的作用，其中 BBB 破坏起着重要作用。
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