BMX Represses Thrombin-PAR1-Mediated Endothelial Permeability and Vascular Leakage During Early Sepsis
BMX Represses Thrombin-PAR1-Mediated Endothelial Permeability and Vascular Leakage During Early Sepsis
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BMX 抑制早期脓毒症期间凝血酶 PAR1 介导的内皮通透性和血管渗漏
DOI：
10.1161/circresaha.119.315769
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发表时间：
2020-02-14
影响因子：
20.1
通讯作者：
Min Wang
中科院分区：
文献类型：
--
作者：
Li Zhao;Yin Mingzhu;Min Wang
原理：BMX（X染色体上的骨髓激酶）在小鼠胚胎期至成年期的动脉内皮中高度表达。它也在微血管和神经系统中表达，以响应病理刺激。然而，它在内皮通透性和sepsis中的作用仍然未知。Objective：我们的目的是描绘BMX在凝血酶介导的内皮通透性和血管渗漏中的功能，在盲肠结扎和穿刺models.Methods和结果：盲肠结扎和穿刺模型应用于WT（野生型）和BMX-KO（BMX全球敲除）小鼠诱导脓毒症。同时采用细胞-基质电阻抗传感法检测体外跨内皮电阻，改良Miles法检测体内血管渗漏。我们发现，BMX的损失造成了早期盲肠结扎和穿刺诱导的脓毒症肺损伤和炎症。破坏BMX增加凝血酶介导的小鼠和培养的内皮细胞的渗透性2至3倍。BMX在巨噬细胞、中性粒细胞、血小板和肺上皮细胞中的表达与在内皮细胞中的表达相比是不可检测的，这表明内皮功能障碍，而不是白细胞和血小板功能障碍，参与了血管通透性和脓毒症。从机制上讲，生物化学和细胞分析表明，BMX特异性抑制凝血酶-PAR 1（蛋白酶激活受体-1）信号在内皮细胞直接磷酸化PAR 1和促进其内化和失活。重要的是，预处理与选择性PAR 1拮抗剂SCH 79797抢救BMX损失介导的内皮细胞通透性和肺渗漏在早期盲肠结扎和穿刺诱导的sepsis.Conclusions：作为一个负调节PAR 1，BMX促进PAR 1的内化和信号失活，通过PAR 1磷酸化。此外，BMX介导的PAR 1内化减弱内皮通透性，以保护早期脓毒症期间的血管渗漏。
Rationale:BMX (bone marrow kinase on the X chromosome) is highly expressed in the arterial endothelium from the embryonic stage to the adult stage in mice. It is also expressed in microvessels and the lymphatics in response to pathological stimuli. However, its role in endothelial permeability and sepsis remains unknown.Objective:We aimed to delineate the function of BMX in thrombin-mediated endothelial permeability and the vascular leakage that occurs with sepsis in cecal ligation and puncture models.Methods and Results:The cecal ligation and puncture model was applied to WT (wild type) and BMX-KO (BMX global knockout) mice to induce sepsis. Meanwhile, the electric cell-substrate impedance sensing assay was used to detect transendothelial electrical resistance in vitro and, the modified Miles assay was used to evaluate vascular leakage in vivo. We showed that BMX loss caused lung injury and inflammation in early cecal ligation and puncture-induced sepsis. Disruption of BMX increased thrombin-mediated permeability in mice and cultured endothelial cells by 2- to 3-fold. The expression of BMX in macrophages, neutrophils, platelets, and lung epithelial cells was undetectable compared with that in endothelial cells, indicating that endothelium dysfunction, rather than leukocyte and platelet dysfunction, was involved in vascular permeability and sepsis. Mechanistically, biochemical and cellular analyses demonstrated that BMX specifically repressed thrombin-PAR1 (protease-activated receptor-1) signaling in endothelial cells by directly phosphorylating PAR1 and promoting its internalization and deactivation. Importantly, pretreatment with the selective PAR1 antagonist SCH79797 rescued BMX loss-mediated endothelial permeability and pulmonary leakage in early cecal ligation and puncture-induced sepsis.Conclusions:Acting as a negative regulator of PAR1, BMX promotes PAR1 internalization and signal inactivation through PAR1 phosphorylation. Moreover, BMX-mediated PAR1 internalization attenuates endothelial permeability to protect vascular leakage during early sepsis.