Thrombin Contributes to Anti-myeloperoxidase Antibody Positive IgG-Mediated Glomerular Endothelial Cells Activation Through SphK1-S1P-S1PR3 Signaling
Thrombin Contributes to Anti-myeloperoxidase Antibody Positive IgG-Mediated Glomerular Endothelial Cells Activation Through SphK1-S1P-S1PR3 Signaling
复制标题
凝血酶通过 SphK1-S1P-S1PR3 信号传导促进抗髓过氧化物酶抗体阳性 IgG 介导的肾小球内皮细胞激活
DOI：
10.3389/fimmu.2019.00237
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发表时间：
2019-02-15
影响因子：
7.3
通讯作者：
Zhao, Ming-Hui
中科院分区：
文献类型：
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作者：
Sun, Xiao-Jing;Chen, Min;Zhao, Ming-Hui
背景资料：凝血系统的激活在抗神经细胞胞浆抗体（ANCA）相关性血管炎（AAV）的发病机制中起重要作用。凝血过程中产生的凝血酶可通过蛋白酶激活受体1（PAR 1）破坏内皮屏障的完整性。我们的前期研究发现，1-磷酸鞘氨醇（S1 P）通过S1 P受体（S1 PR）依赖性途径参与髓过氧化物酶（MPO）-ANCA阳性IgG诱导的肾小球内皮细胞（GEnC）活化。近年来，S1 P信号被报道参与凝血酶对内皮细胞的作用。本研究探讨凝血酶-PAR和S1 P-S1 PR信号之间的相互作用是否导致MPO-ANCA阳性IgG诱导的GEnC功能障碍。方法：从三个方面分析凝血酶对GEnC激活的影响。首先，观察GEnCs的形态学改变。第二，进行渗透性测定以定量测定GEnC单层活化。第三，测定内皮素-1（ET-1）水平。检测鞘氨醇激酶（SphKs）和S1 PRs的表达水平。此外，使用PAR 1和S1 PR 3的拮抗剂来确定它们的作用。结果：MPO-ANCA阳性IgG刺激后，凝血酶可进一步损伤GEnCs的紧密连接，增加内皮细胞单层通透性，上调内皮素1（ET-1）水平。阻断PAR 1可下调凝血酶加MPO ANCA阳性IgG处理的GEnCs培养上清中ET-1的水平。在凝血酶+MPO-ANCA阳性IgG处理的GEnC中，SphK 1、S1 PR 3的表达水平显著增加。结论：凝血酶通过与SphK 1-S1 P-S1 PR 3信号通路协同作用，增强MPO-ANCA阳性IgG介导的GEnC活化。
Background: Activation of coagulation system plays an important role in antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) pathogenesis. Thrombin, generated during coagulation could disrupt endothelial barrier integrity through protease-activated receptor 1 (PAR1). Our previous study found that sphingosine-1-phosphate (S1P) contributed to myeloperoxidase (MPO)-ANCA-positive IgG-induced glomerular endothelial cell (GEnC) activation through a S1P receptor (S1PR)-dependent route. In recent years, S1P signaling was reported to be involved in thrombin effects on endothelial cells. This current study investigated whether the interaction between thrombin-PAR and S1P-S1PR signaling contributed to MPO-ANCA-positive IgG-induced GEnC dysfunction.Methods: The effect of thrombin on GEnC activation was analyzed from three aspects. First, morphological alteration of GEnCs was observed. Second, permeability assay was performed to determine GEnC monolayer activation quantitatively. Third, endothelin-1 (ET-1) levels were measured. Expression levels of sphingosine kinases (SphKs) and S1PRs were detected. In addition, antagonists of PAR1 and S1PR3 were employed to determine their roles. Eventually, PAR1 and tissue factor (TF) expression levels as well as TF procoagulant activity were analyzed.Results: Thrombin induced further damage of tight junction, increase in endothelial monolayer permeability as well as upregulation of ET-1 levels in GEnCs stimulated with MPO-ANCA-positive IgG. Blocking PAR1 downregulated ET-1 levels in the supernatants of GEnCs treated by thrombin plusMPO-ANCA-positive IgG. Expression levels of SphK1, S1PR3 increased significantly in GEnCs treated with thrombin plus MPO-ANCA-positive IgG. S1P upregulated PAR1 and TF expression, and enhanced procoagulant activity of TF in MPO-ANCA-positive IgG-stimulated GEnCs.Conclusion: Thrombin synergized with SphK1-S1P-S1PR3 signaling pathway to enhance MPO-ANCA-positive IgG-mediated GEnC activation.