BACKGROUND Lymphatic collecting vessels in the kidney are critical in clearing interstitial fluid, macromolecules, and infiltrating immune cells. Dysfunction of the lymphatic vessels can disrupt this process and exacerbate injury-associated inflammation in many disease conditions. We previously found that sodium accumulates within the kidney interstitium during proteinuric kidney injury and elevated sodium environments stimulate isolevuglandin production in antigen-presenting cells, stimulating T cells, and modulating inflammatory responses. In the present study, we investigated whether proteinuric injury increases production of isolevuglandin-adduct formation in antigen-presenting cells, their effects on lymphatic endothelial cells (LECs), and the role of the ET-3 (endothelin-3)/ETBR (endothelin type B receptor) on lymphatic vessel function. METHODS We used a mouse model of nephrotoxin-induced proteinuric injury to show that proteinuric injury expanded the kidney lymphatic network and to immunophenotype the infiltrating immune cells. To determine mechanisms, we analyzed the interaction of migratory immune cells and LECs using an in vitro transwell migration assay, bulk RNA sequencing, and flow cytometric analysis. To determine the effect of ET-3/ETBR axis on lymphatic vessel contractility, we analyzed microdissected lymphangions utilizing a vessel perfusion chamber. RESULTS We found that animals with proteinuric injury have increased kidney lymphangiogenesis, isolevuglandin-producing dendritic cells, and IFN (interferon)-γ-producing CD4+T cells. The sodium avid environment present in kidney injury enhances the interaction between LECs and migratory antigen-presenting cells and LEC production of isolevuglandin-adducts. Elevated sodium environment-induced isolevuglandin-adduct formation facilitates the ET-3/ETBR communication between LECs and dendritic cells. In addition, the ET-3/ETBR axis modulates lymphatic collecting vessel pumping dynamics. CONCLUSIONS These findings reveal a novel mechanism linking the isolevuglandin-mediated ET-3/ETBR axis with LECs and infiltrating dendritic cells. ET-3/ETBR signaling in lymphatic vessel dynamics is a novel pathogenic component and a possible therapeutic target in kidney disease.

中文翻译：

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ET-3/ETBR 介导 Na+ 激活的免疫信号传导和肾脏淋巴动力学。


背景 肾脏中的淋巴集合血管对于清除间质液、大分子和浸润免疫细胞至关重要。在许多疾病情况下，淋巴管功能障碍会破坏这一过程并加剧与损伤相关的炎症。我们之前发现，在蛋白尿性肾损伤期间钠在肾间质内积累，升高的钠环境刺激抗原呈递细胞中异亮腺素的产生，刺激 T 细胞，并调节炎症反应。在本研究中，我们研究了蛋白尿损伤是否会增加抗原呈递细胞中异乙酰腺素加合物形成的产生，它们对淋巴内皮细胞 （LEC） 的影响，以及 ET-3 （内皮素-3）/ETBR （内皮素 B 型受体） 对淋巴管功能的作用。方法 我们使用肾毒素诱导的蛋白尿损伤小鼠模型来表明蛋白尿损伤扩大了肾淋巴网络并对浸润免疫细胞进行免疫表型分析。为了确定机制，我们使用体外 transwell 迁移测定、大量 RNA 测序和流式细胞术分析分析了迁移免疫细胞和 LEC 的相互作用。为了确定 ET-3/ETBR 轴对淋巴管收缩力的影响，我们利用血管灌注室分析了显微解剖的淋巴管。结果我们发现，蛋白尿损伤的动物肾淋巴管生成、产生异亮腺素的树突状细胞和产生 IFN （干扰素）γ CD4+T 细胞增加。肾损伤中存在的钠亲和环境增强了 LEC 与迁移抗原呈递细胞之间的相互作用以及 LEC 产生异亮腺素加合物。 钠环境诱导的异亮腺素加合物形成升高促进了 LEC 和树突状细胞之间的 ET-3/ETBR 通讯。此外，ET-3/ETBR 轴调节淋巴收集血管泵血动力学。结论 这些发现揭示了一种将异左兰素介导的 ET-3/ETBR 轴与 LECs 和浸润树突状细胞联系起来的新机制。淋巴管动力学中的 ET-3/ETBR 信号转导是一种新型致病成分，是肾脏疾病的可能治疗靶点。