Histidine-rich glycoprotein ameliorates endothelial barrier dysfunction through regulation of NF-κB and MAPK signal pathway
Histidine-rich glycoprotein ameliorates endothelial barrier dysfunction through regulation of NF-κB and MAPK signal pathway
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DOI：
10.1111/bph.14711
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发表时间：
2019-08-01
影响因子：
7.3
通讯作者：
Nishibori, Masahiro
中科院分区：
文献类型：
--
作者：
Gao, Shangze;Wake, Hidenori;Nishibori, Masahiro
背景和目的微血管屏障破坏是脓毒症的标志，与脓毒症诱导的多器官衰竭相关。富含组氨酸的糖蛋白（HRG）是一种75 kDa的血浆蛋白，经证实可通过调节细胞形状、中性粒细胞中的自发ROS产生以及中性粒细胞与血管内皮细胞的粘附来改善脓毒症小鼠的存活率。我们研究了HRG在体内外LPS/TNF-α诱导的内皮细胞屏障功能障碍中的作用及其可能的机制，以阐明HRG在脓毒症中的确切作用。实验方法EA.hy926内皮细胞在用LPS/TNF-α刺激之前用HRG或人血清白蛋白预处理。本研究采用多种生物化学方法探讨HRG保护血管内皮屏障功能的分子机制。免疫组化结果显示HRG通过抑制细胞骨架重组、VE-钙粘蛋白和β-连环蛋白的丢失、粘着斑激酶降解和LPS/TNF-α诱导的细胞脱离来维持内皮单层的完整性。HRG还抑制LPS/TNF-α诱导的内皮细胞分泌细胞因子，这与NF-κ B活化减少有关。此外，HRG有效地防止了LPS/TNF-α诱导的体外和体内毛细血管通透性增加。最后，Western blot结果表明HRG抑制MAPK家族的磷酸化和RhoA的激活，这两种蛋白质主要参与细胞骨架重组和屏障通透性的调节。结论：HRG对血管屏障功能具有保护作用，其机制可能与抑制MAPK家族和Rho激活有关。
Background and Purpose Microvascular barrier breakdown is a hallmark of sepsis that is associated with sepsis-induced multiorgan failure. Histidine-rich glycoprotein (HRG) is a 75-kDa plasma protein that was demonstrated to improve the survival of septic mice through regulation of cell shape, spontaneous ROS production in neutrophils, and adhesion of neutrophils to vascular endothelial cells. We investigated HRG's role in the LPS/TNF-alpha-induced barrier dysfunction of endothelial cells in vitro and in vivo and the possible mechanism, to clarify the definitive roles of HRG in sepsis. Experimental Approach EA.hy 926 endothelial cells were pretreated with HRG or human serum albumin before stimulation with LPS/TNF-alpha. A variety of biochemical assays were applied to explore the underlying molecular mechanisms on how HRG protected the barrier function of vascular endothelium. Key Results Immunostaining results showed that HRG maintains the endothelial monolayer integrity by inhibiting cytoskeleton reorganization, losses of VE-cadherin and beta-catenin, focal adhesion kinase degradation, and cell detachment induced by LPS/TNF-alpha. HRG also inhibited the cytokine secretion from endothelial cells induced by LPS/TNF-alpha, which was associated with reduced NF-kappa B activation. Moreover, HRG effectively prevented the LPS/TNF-alpha-induced increase in capillary permeability in vitro and in vivo. Finally, Western blot results demonstrated that HRG prevented the phosphorylation of MAPK family and RhoA activation, which are involved mainly in the regulation of cytoskeleton reorganization and barrier permeability. Conclusions and Implications Taken together, our results demonstrate that HRG has protective effects on vascular barrier function in vitro and in vivo, which may be due to the inhibition of MAPK family and Rho activation.