State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China.
Zhejiang Provincial Key Laboratory of Pancreatic Disease, the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China.
Hepatol Commun. 2023 Sep 15;7(10). doi: 10.1097/HC9.0000000000000257. eCollection 2023 Oct 1.
Acute-on-chronic liver failure (ACLF) is an acute decompensated syndrome based on chronic liver disease, while neutrophil recruitment is the most critical early step. C-X-C motif chemokine ligand 1 (CXCL1), a cytokine that recruits neutrophils, was significantly upregulated in both ACLF mice and patients with ACLF. This present study aims to explore the role of CXCL1 in the pathogenesis of ACLF.
We established an ACLF mouse model induced by carbon tetrachloride, lipopolysaccharide, and D-galactosamine, and used adeno-associated virus to achieve overexpression and knockdown of Cxcl1. We employed mass cytometry, flow cytometry, multiplex cytokine and chemokine analysis, Western blot, and reactive oxygen species (ROS) detection in mice blood and liver. ACLF patients (n = 10) and healthy controls (n = 5) were included, and their liver samples were stained using multiplex immunohistochemistry techniques.
CXCL1 was significantly elevated in both ACLF mice and patients. CXCL1 recruits neutrophils by binding to the C-X-C motif chemokine receptor 2 on the surface of neutrophils, affects ACLF prognosis by generating ROS and mitochondrial depolarization and modulating caspase3-related apoptotic pathways. We found that the knockdown of CXCL1 attenuated the infiltration of neutrophils in the mouse liver, reduced the expression of inflammatory cytokines, and also significantly downregulated ROS production and caspase3-related hepatocyte apoptosis, thereby ameliorating the liver injury of ACLF.
CXCL1 is a core player in the mobilization of neutrophils in ACLF, and the knockdown of Cxcl1 improves neutrophil infiltration, reduces ROS levels, and reduces hepatocyte apoptosis, thereby attenuating inflammation and liver injury in ACLF. Our results revealed a previously unknown link between CXCL1-induced neutrophil recruitment and ACLF, providing evidencing for potential therapies targeting ACLF.
慢加急性肝衰竭（ACLF）是基于慢性肝病的急性失代偿综合征，而中性粒细胞募集是最关键的早期步骤。C-X-C 基序趋化因子配体 1（CXCL1）是一种募集中性粒细胞的细胞因子，在 ACLF 小鼠和 ACLF 患者中均显著上调。本研究旨在探讨 CXCL1 在 ACLF 发病机制中的作用。
我们建立了由四氯化碳、脂多糖和 D-半乳糖胺诱导的 ACLF 小鼠模型，并使用腺相关病毒实现 Cxcl1 的过表达和敲低。我们在小鼠血液和肝脏中使用质谱流式细胞术、流式细胞术、多重细胞因子和趋化因子分析、Western blot 和活性氧（ROS）检测来研究 Cxcl1 的作用。纳入了 10 名 ACLF 患者和 5 名健康对照者，并用多重免疫组化技术对他们的肝组织样本进行染色。
在 ACLF 小鼠和患者中，CXCL1 均显著升高。CXCL1 通过与中性粒细胞表面的 C-X-C 基序趋化因子受体 2 结合来募集中性粒细胞，通过产生 ROS 和线粒体去极化以及调节 caspase3 相关凋亡途径来影响 ACLF 的预后。我们发现，敲低 CXCL1 可减轻小鼠肝脏中性粒细胞的浸润，降低炎症细胞因子的表达，同时还显著下调 ROS 产生和 caspase3 相关的肝细胞凋亡，从而改善 ACLF 的肝损伤。
CXCL1 是 ACLF 中中性粒细胞动员的核心分子，敲低 Cxcl1 可减少中性粒细胞浸润，降低 ROS 水平，并减少肝细胞凋亡，从而减轻 ACLF 的炎症和肝损伤。我们的研究结果揭示了 CXCL1 诱导的中性粒细胞募集与 ACLF 之间的一个以前未知的联系，为靶向 ACLF 的潜在治疗提供了证据。
