Toll-Like Receptor 3 Signaling Contributes to Regional Neutrophil Recruitment in Cultured Human Glomerular Endothelial Cells
Toll-Like Receptor 3 Signaling Contributes to Regional Neutrophil Recruitment in Cultured Human Glomerular Endothelial Cells
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DOI：
10.1159/000489507
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发表时间：
2018-05
期刊：
影响因子：
2.5
通讯作者：
Qiang Liu;T. Imaizumi;S. Kawaguchi;Tomomi Aizawa;T. Matsumiya;Shojiro Watanabe;K. Tsugawa;H. Yoshida;K. Tsuruga;K. Joh;H. Kijima;Hiroshi Tanaka
中科院分区：
文献类型：
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作者：
Qiang Liu;T. Imaizumi;S. Kawaguchi;Tomomi Aizawa;T. Matsumiya;Shojiro Watanabe;K. Tsugawa;H. Yoshida;K. Tsuruga;K. Joh;H. Kijima;Hiroshi Tanaka
背景：鉴于中性粒细胞募集在肾小球肾炎发病机制中的重要性，具有代表性的中性粒细胞趋化因子C-X-C基序趋化因子1/Groα和肾小球内皮细胞黏附分子E-选择素在肾小球肾炎的发生发展中起关键作用。内皮细胞Toll样受体3(TLR3)被认为通过先天免疫参与炎症反应。然而，内皮细胞TLR3信号在中性粒细胞趋化因子和黏附分子表达中的作用尚不清楚。因此，我们的目标是研究这个问题。方法：采用多聚肌苷-多胞苷(PolyIC)处理正常人肾小管上皮细胞，采用实时定量逆转录聚合酶链式反应、Western blotting和酶联免疫吸附试验检测CXCL1和E-选择素的表达。为了进一步阐明Poly IC诱导的信号通路，我们对TLR3、干扰素-β、核因子-κB p65和干扰素调节因子3进行了RNA干扰。我们还使用免疫荧光技术检测了新月体和非新月体紫癜性肾炎(PN)患者活检标本中CXCL1的内皮表达。结果：我们发现TLR3的激活诱导了CXCL1和E-选择素的内皮表达，并且参与了TLR3、NF-κB、IRF3和干扰素-β的表达。在新月体PN患者的活检标本中观察到内皮细胞CXCL1的强烈表达。结论：这些发现支持肾小球天然免疫在肾小球肾炎发病机制中的作用。因此，干预肾小球TLR3信号可能是未来治疗肾小球肾炎的合适治疗策略。
Background: Given the importance of neutrophil recruitment in the pathogenesis of glomerulonephritis (GN), the representative neutrophil chemoattractant C-X-C motif chemokine 1 (CXCL1)/GROα and the adhesion molecule E-selectin in glomerular endothelial cells (GECs) play a pivotal role in the development of GN. Endothelial Toll-like receptor 3 (TLR3) is thought to be involved in the inflammatory response via innate immunity. However, the role of endothelial TLR3 signaling in the expression of neutrophil chemoattractants and adhesion molecules remains to be elucidated. Thus, we aimed to examine this issue. Methods: We treated normal human GECs with polyinosinic-polycytidylic acid (poly IC), an authentic double-stranded RNA, and analyzed the expressions of CXCL1 and E-selectin using quantitative real-time reverse transcription-polymerase chain reaction, western blotting, and enzyme-linked immunosorbent assay. To further elucidate the poly IC-induced signaling pathway, we subjected the cells to RNA interference against TLR3, interferon (IFN)-β, nuclear factor (NF)-κB p65, and IFN regulatory factor (IRF) 3. We also used immunofluorescence to examine the endothelial expression of CXCL1 in biopsy specimens from patients with crescentic and non-crescentic purpura nephritis (PN). Results: We found that the activation of TLR3 induced the endothelial expression of CXCL1 and E-selectin, and that this involved TLR3, NF-κB, IRF3, and IFN-β. Intense endothelial CXCL1 expression was observed in biopsy specimens from patients with crescentic PN. Conclusion: These findings support a role for glomerular antiviral innate immunity in the pathogenesis of GN. Intervention of glomerular TLR3 signaling may therefore be a suitable therapeutic strategy for treating GN in the future.