Yes-associated protein (YAP) signaling regulates lipopolysaccharide-induced tissue factor expression in human endothelial cells
Yes-associated protein (YAP) signaling regulates lipopolysaccharide-induced tissue factor expression in human endothelial cells
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Yes相关蛋白（YAP）信号调节人内皮细胞中脂多糖诱导的组织因子表达
DOI：
10.1016/j.surg.2015.12.008
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发表时间：
2016
期刊：
影响因子：
3.8
通讯作者：
Huan Jingning
中科院分区：
文献类型：
--
作者：
Yi Lei;Huang Xiaoqin;Guo Feng;Zhou Zengding;Dou Yi;Huan Jingning
脓毒症急性肺损伤（acute lung injury，ALI）以纤维蛋白沉积为特征，是局部凝血功能激活的标志。组织因子（TF）表达于肺微血管，是脓毒症中凝血和ALI的关键起始因子。然而，脂多糖（LPS）诱导内皮细胞（EC）表达TF的分子机制尚未确定。在这项研究中，我们牵连的Rho相关蛋白激酶（ROCK）/是相关蛋白（雅普）/早期生长反应（Egr-1）信号通路在LPS诱导的TF表达在体外和脓毒症诱导的ALIinvivo.MethodsHuman脐静脉内皮细胞与LPS孵育预处理或不与ROCK抑制剂Y-27632，一个雅普小，干扰RNA（siRNA）和Egr-1 siRNA。Western blot检测ROCK、雅普和Egr-1信号转导蛋白的表达。通过免疫荧光测定分析LPS诱导的雅普活化。此外，我们intraperitheally注射雅普siRNA评估脓毒性ALI小鼠苏木素和伊红staining.ResultsLPS迅速诱导ROCK激活和增加TF表达EC。LPS诱导雅普穿梭于内皮细胞核内，并通过激活ROCK与Egr-1结合。此外，ROCK失活、雅普敲低和Egr-1缺失均能抑制LPS诱导的TF表达，提示LPS诱导的TF表达与ROCK/雅普/Egr-1信号通路密切相关。最后，气管内注射雅普siRNA减轻脓毒症小鼠的肺损伤。ConclusionThis study not only suggests that ROCK/雅普/Egr-1 signaling regulates TF expression after stimulation with LPS in ECs，but it also suggests，LPS诱导的雅普signaling activation plays an important role in septic ALI in mice.我们的研究结果为TF表达的致病机制提供了新的见解，TF表达与脓毒症ALI密切相关，雅普信号传导被认为是脓毒症条件下治疗干预的新靶点。
BackgroundSepsis-induced acute lung injury (ALI) is characterized by fibrin deposition, which indicates the local activation of coagulation. Tissue factor (TF), expressed in the pulmonary microvasculature, acts as a critical initiator of blood coagulation and ALI in sepsis. The molecular mechanism of lipopolysaccharide (LPS)-induced TF expression in endothelial cells (ECs), however, has not been determined. In this study, we implicate the Rho-associated protein kinase (ROCK)/Yes associated protein (YAP)/early growth response (Egr-1) signaling pathway in LPS-induced TF expression in vitro and in sepsis-induced ALI in vivo.MethodsHuman umbilical vein ECs incubated with LPS were pretreated with or without the ROCK inhibitor Y-27632, a YAP small, interfering RNA (siRNA) and an Egr-1 siRNA. ROCK, YAP and Egr-1 signaling–induced protein expression was investigated by Western blot. The LPS-induced activation of YAP was analyzed by an immunofluorescent assay. Furthermore, we intratracheally injected YAP siRNA to assess septic ALI in mice by hematoxylin and eosin staining.ResultsLPS rapidly induced ROCK activation and increased TF expression in ECs. LPS caused YAP shuttling into the nuclei of ECs and combined with Egr-1 via the activation of ROCK. Furthermore, the LPS-mediated TF expression increase was prevented by ROCK inactivation, YAP knockdown and Egr-1 depletion, suggesting that LPS-induced TF expression is closely associated with the ROCK/YAP/Egr-1 signaling pathway in ECs. Finally, an intratracheal injection of YAP siRNA relieved lung injury in septic mice.ConclusionThis study not only suggests that ROCK/YAP/Egr-1 signaling regulates TF expression after stimulation with LPS in ECs, but it also indicates that LPS-induced activation of YAP signaling plays an important role in septic ALI in mice. Our findings provide a new insight into the pathogenic mechanism of TF expression, which is closely linked to septic ALI, and YAP signaling is considered to be a novel target for therapeutic intervention under septic conditions.