Selective inhibition of brain endothelial Rho-kinase-2 provides optimal protection of an in vitro blood-brain barrier from tissue-type plasminogen activator and plasmin.
Selective inhibition of brain endothelial Rho-kinase-2 provides optimal protection of an in vitro blood-brain barrier from tissue-type plasminogen activator and plasmin.
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DOI：
10.1371/journal.pone.0177332
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Medcalf RL
中科院分区：
文献类型：
--
作者：
Niego B;Lee N;Larsson P;De Silva TM;Au AE;McCutcheon F;Medcalf RL
Rho激酶（ROCK）抑制剂广泛用于心血管疾病，可在溶栓过程中保护血脑屏障（BBB）免受rt-PA诱导的损伤。虽然非选择性ROCK抑制剂如法舒地尔与rt-PA一起使用可能受到可能的副作用和阻断脑内皮细胞（BEC）中有害rt-PA活性的能力不足的阻碍，但选择性ROCK-2抑制剂可以克服这些限制。在这里，我们研究了ROCK-2在主要脑细胞中的表达，并比较了法舒地尔和KD 025（一种选择性ROCK-2抑制剂）在体外人体模型中减弱rt-PA诱导的BBB损伤的能力。ROCK-2在所有人类和小鼠脑细胞类型中相对于ROCK-1高度表达，并且与神经元相比，在啮齿动物脑内皮细胞和星形胶质细胞中特别富集。在常氧条件下，KD 025在衰减rt-PA和纤溶酶原诱导的BBB渗透方面比法舒地尔更有效，但在卒中样条件下尤其如此。重要的是，只有KD 025，而不是法舒地尔，能够阻止rt-PA依赖性的渗透性增加，形态学变化和紧密连接降解分离的BEC。选择性ROCK-2抑制进一步减少了星形胶质细胞中rt-PA触发的肌球蛋白磷酸化、形状改变和基质金属蛋白酶激活。这些发现强调了ROCK-2是rt-PA导致BBB损伤和脑内皮损伤的关键亚型，以及KD 025在溶栓过程中最佳保护BBB的潜力。
Rho-kinase (ROCK) inhibition, broadly utilised in cardiovascular disease, may protect the blood-brain barrier (BBB) during thrombolysis from rt-PA-induced damage. While the use of nonselective ROCK inhibitors like fasudil together with rt-PA may be hindered by possible hypotensive side-effects and inadequate capacity to block detrimental rt-PA activity in brain endothelial cells (BECs), selective ROCK-2 inhibition may overcome these limitations. Here, we examined ROCK-2 expression in major brain cells and compared the ability of fasudil and KD025, a selective ROCK-2 inhibitor, to attenuate rt-PA-induced BBB impairment in an in vitro human model. ROCK-2 was highly expressed relative to ROCK-1 in all human and mouse brain cell types and particularly enriched in rodent brain endothelial cells and astrocytes compared to neurons. KD025 was more potent than fasudil in attenuation of rt-PA- and plasminogen-induced BBB permeation under normoxia, but especially under stroke-like conditions. Importantly, only KD025, but not fasudil, was able to block rt-PA-dependent permeability increases, morphology changes and tight junction degradation in isolated BECs. Selective ROCK-2 inhibition further diminished rt-PA-triggered myosin phosphorylation, shape alterations and matrix metalloprotease activation in astrocytes. These findings highlight ROCK-2 as the key isoform driving BBB impairment and brain endothelial damage by rt-PA and the potential of KD025 to optimally protect the BBB during thrombolysis.