EphrinA1/EphA2 Promotes Epithelial Hyperpermeability Involving in Lipopolysaccharide-induced Intestinal Barrier Dysfunction
EphrinA1/EphA2 Promotes Epithelial Hyperpermeability Involving in Lipopolysaccharide-induced Intestinal Barrier Dysfunction
复制标题
EphrinA1/ephA2 促进上皮通透性过高，参与脂多糖诱导的肠道屏障功能障碍
DOI：
10.5056/jnm19095
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发表时间：
2020-07-01
影响因子：
3.4
通讯作者：
Hou, Xiaohua
中科院分区：
文献类型：
--
作者：
Chen, Yuhua;Zhang, Lei;Hou, Xiaohua
背景/目的脂多糖（Lipopolysaccharide，LPS）是引起各种肠道和胃肠外疾病的黏膜和全身炎症的关键因素，最初可破坏上皮屏障功能。推测EphrinA 1/ephA 2由于其在相邻细胞之间的“排斥性相互作用”而增加上皮渗透性。本研究旨在探讨ephrinA 1/ephA 2在LPS诱导的上皮细胞通透性增高中的作用。评估了屏障功能，包括紧密连接蛋白（闭合蛋白和封闭蛋白-1）的表达、跨上皮电阻和对大分子（异硫氰酸荧光素标记的荧光葡聚糖4 kDa [FD 4]）的渗透性。此外，通过用ephrinA 1-Fc嵌合体刺激ephA 2来定量ephrinA 1/ephA 2的表达和磷酸化，并证实其在上皮屏障破坏过程中的作用用ephA 2-Fc嵌合体灭活ephA 2（ephA 2-Fc）或ephA 2单克隆抗体（ephA2-mAb），以及抑制细胞外信号调节激酶1/2（ERK 1/2），结果LPS可诱导内皮细胞屏障功能障碍，表现为occludin和claudin-1表达减少，跨上皮电阻降低，FD 4通透性增加，伴随着ephrinA 1/ephA 2途径的上调和ephA 2受体的磷酸化。此外，ephA 2-Fc和ephA 2- mab改善LPS诱导的上皮细胞通透性过高，这也被PD 98059抑制。EphrinA 1-Fc可通过促进ERK 1/2磷酸化而导致Caco 2单层上皮细胞渗漏，而EphA 2-mAb和PD 98059可明显阻断EphrinA 1/EphA 2通过ERK 1/2依赖性途径促进上皮细胞通透性增高，参与LPS诱导的肠屏障功能障碍。
Background/AimsLipopolysaccharide (LPS) is the key factor inducing mucosal and systemic inflammation in various intestinal and parenteral diseases, which could initially disrupt the epithelial barrier function. EphrinA1/ephA2 is speculated to increase the epithelial permeability for its "repulsive interaction" between adjacent cells. This study aim to investigate the role of ephrinA1/ephA2 in LPS-induced epithelial hyperpermeability.MethodsIn vivo model challenged with oral LPS in C57BL/6 mice and in vitro model exposed to LPS in Caco2 monolayer were established. The barrier function was assessed including expression of tight junction proteins (occludin and claudin-1), transepithelial electrical resistance, and permeability to macromolecules (fluorescein isothiocyanate-labeled fluorescent dextran 4 kDa [FD4]). Moreover, the expression and phosphorylation of ephrinA1/ephA2 were quantified, and its roles in the process of epithelial barrier disruption were confirmed via stimulating ephA2 with ephrinA1-Fc chimera (ephrinA1-Fc) and inactivating ephA2 with ephA2-Fc chimera (ephA2-Fc), or ephA2 monoclonal antibody (ephA2-mab), as well as inhibiting extracellular signal-regulated kinase 1/2 (ERK1/2) with PD98059.ResultsLPS induced significant barrier dysfunction with dismissed occludin and claudin-1 expression, reduced transepithelial electrical resistance and increased FD4 permeability, accompanied by upregulated ephrinA1/ephA2 pathway and phosphorylation of ephA2 receptor. Furthermore, ephA2-Fc, and ephA2- mab ameliorated LPS-induced epithelial hyperpermeability, which was also inhibited by PD98059. Additionally, ephrinA1-Fc led to apparent epithelial leakage in Caco2 monolayer by promoting the phosphorylation of ERK1/2, which could be obviously blocked by ephA2-mab and PD98059.ConclusionEphrinA1/ephA2 promotes epithelial hyperpermeability with an ERK1/2-dependent pathway, which involves in LPS-induced intestinal barrier dysfunction.