Czepielewski Rafael S, Jaeger Natália, Marques Pedro E, Antunes Maísa M, Rigo Maurício M, Alvarenga Débora M, Pereira Rafaela V, da Silva Rodrigo D, Lopes Tiago G, da Silva Vinícius D, Porto Bárbara N, Menezes Gustavo B, Bonorino Cristina
Laboratório de Imunologia Celular e Molecular, Instituto de Pesquisas Biomédicas (IPB), Porto Alegre, RS, Brazil.
Departamento de Bioquímica e Imunologia, Laboratório de Imunofarmacologia, UFMG, Belo Horizonte, MG, Brazil.
Eur J Immunol. 2017 Apr;47(4):646-657. doi: 10.1002/eji.201646394.
Drug-induced liver injury (DILI) is a major cause of acute liver failure (ALF), where hepatocyte necrotic products trigger liver inflammation, release of CXC chemokine receptor 2 (CXCR2) ligands (IL-8) and other neutrophil chemotactic molecules. Liver infiltration by neutrophils is a major cause of the life-threatening tissue damage that ensues. A GRPR (gastrin-releasing peptide receptor) antagonist impairs IL-8-induced neutrophil chemotaxis in vitro. We investigated its potential to reduce acetaminophen-induced ALF, neutrophil migration, and mechanisms underlying this phenomenon. We found that acetaminophen-overdosed mice treated with GRPR antagonist had reduced DILI and neutrophil infiltration in the liver. Intravital imaging and cell tracking analysis revealed reduced neutrophil mobility within the liver. Surprisingly, GRPR antagonist inhibited CXCL2-induced migration in vivo, decreasing neutrophil activation through CD11b and CD62L modulation. Additionally, this compound decreased CXCL8-driven neutrophil chemotaxis in vitro independently of CXCR2 internalization, induced activation of MAPKs (p38 and ERK1/2) and downregulation of neutrophil adhesion molecules CD11b and CD66b. In silico analysis revealed direct binding of GRPR antagonist and CXCL8 to the same binding spot in CXCR2. These findings indicate a new potential use for GRPR antagonist for treatment of DILI through a mechanism involving adhesion molecule modulation and possible direct binding to CXCR2.
药物性肝损伤（DILI）是急性肝衰竭（ALF）的主要原因，其中肝细胞坏死产物引发肝脏炎症，释放CXC趋化因子受体2（CXCR2）配体（IL-8）和其他中性粒细胞趋化分子。中性粒细胞浸润肝脏是随后发生的危及生命的组织损伤的主要原因。一种胃泌素释放肽受体（GRPR）拮抗剂在体外会损害IL-8诱导的中性粒细胞趋化作用。我们研究了其减轻对乙酰氨基酚诱导的ALF、中性粒细胞迁移以及这种现象背后机制的潜力。我们发现，用GRPR拮抗剂治疗的对乙酰氨基酚过量小鼠的DILI和肝脏中的中性粒细胞浸润有所减轻。活体成像和细胞追踪分析显示肝脏内中性粒细胞的迁移率降低。令人惊讶的是，GRPR拮抗剂在体内抑制CXCL2诱导的迁移，通过调节CD11b和CD62L减少中性粒细胞活化。此外，该化合物在体外独立于CXCR2内化降低CXCL8驱动的中性粒细胞趋化作用，诱导丝裂原活化蛋白激酶（p38和ERK1/2）活化并下调中性粒细胞黏附分子CD11b和CD66b。计算机模拟分析显示GRPR拮抗剂和CXCL8直接结合到CXCR2中的同一结合位点。这些发现表明GRPR拮抗剂通过涉及黏附分子调节和可能直接结合CXCR2的机制治疗DILI有新的潜在用途。
