Endothelial NF-κB Blockade Abrogates ANCA-Induced GN
Endothelial NF-κB Blockade Abrogates ANCA-Induced GN
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DOI：
10.1681/asn.2016060690
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发表时间：
2017-11-01
影响因子：
13.6
通讯作者：
Kettritz, Ralph
中科院分区：
文献类型：
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作者：
Choi, Mira;Schreiber, Adrian;Kettritz, Ralph
ANCA相关性血管炎（AAV）是一种高度炎症性疾病，其中ANCA激活的中性粒细胞与内皮细胞相互作用，导致坏死性血管炎。我们验证了内皮NF-κ B B介导坏死性新月体肾炎（NCGN）并提供特异性治疗靶点的假设。对来自先前检查的鼠NCGN疾病模型的肾脏的再分析揭示了受影响肾脏中的NF-κ B活化，主要为p50/p65异源二聚体，并且NF-κ B依赖性肿瘤坏死因子α（TNF-α）的肾脏表达增加。NF-κ B活化与新月体形成呈正相关，核磷酸化p65染色显示受影响肾小球中表达CD 31的内皮细胞（EC）内NF-κ B活化。因此，我们在体外研究了ANCA对中性粒细胞/EC共培养物中NF-κ B活化的影响。ANCA不能激活致敏的人中性粒细胞中的NF-κ B B，但ANCA刺激的致敏的中性粒细胞激活了EC中的NF-κ B B，至少部分是通过TNF-α的释放。这种作用增加了NF-κ B调节的白细胞介素-8的内皮基因转录和蛋白质产生。此外，内皮细胞NF-κ B的上调促进了中性粒细胞与EC单层的粘附，这种作用可被特异性IKK β抑制剂抑制。在小鼠NCGN模型中，预防性应用E-选择素靶向免疫脂质体（包装有p65 siRNA）下调内皮NF-κ B，显著减少了尿液异常、肾髓样细胞流入和NCGN。AAV患者肾小球内皮磷酸化p65染色增加表明NF-κ B在人NCGN中也被激活。我们认为ANCA刺激的中性粒细胞激活内皮NF-κ B B，这有助于NCGN，并提供了一个潜在的治疗靶点在AAV。
ANCA-associated vasculitis (AAV) is a highly inflammatory condition in which ANCA-activated neutrophils interact with the endothelium, resulting in necrotizing vasculitis. We tested the hypothesis that endothelial NF-kappa B mediates necrotizing crescentic GN(NCGN) and provides a specific treatment target. Reanalysis of kidneys from previously examined murine NCGN disease models revealed NF-kappa B activation in affected kidneys, mostly as a p50/p65 heterodimer, and increased renal expression of NF-kappa B-dependent tumor necrosis factor a (TNF-alpha). NF-kappa B activation positively correlated with crescent formation, and nuclear phospho-p65 staining showedNF-kappa B activation within CD31-expressing endothelial cells (ECs) in affected glomeruli. Therefore, we studied the effect of ANCA on NF-kappa B activation in neutrophil/EC cocultures in vitro. ANCA did not activate NF-kappa B in primed human neutrophils, but ANCA-stimulated primed neutrophils activated NF-kappa B in ECs, at least in part via TNF-alpha release. This effect increased endothelial gene transcription and protein production of NF-kappa B-regulated interleukin-8. Moreover, upregulation of endothelial NF-kappa B promoted neutrophil adhesion to EC monolayers, an effect that was inhibited by a specific IKK beta inhibitor. In a murine NCGN model, prophylactic application of E-selectin-targeted immunolipo-somes packed with p65 siRNA to downregulate endothelial NF-kappa B significantly reduced urine abnormalities, renal myeloid cell influx, and NCGN. Increased glomerular endothelial phospho-p65 staining in patients with AAV indicated that NF-kappa B is activated in human NCGN also. We suggest that ANCA-stimulated neutrophils activate endothelial NF-kappa B, which contributes to NCGN and provides a potential therapeutic target in AAV.