Department of General Surgery, Xuanwu Hospital, Capital Medical University, Beijing, PR China; Clinical Center for Acute Pancreatitis, Capital Medical University, Beijing, PR China.
Department of General Surgery, Xuanwu Hospital, Capital Medical University, Beijing, PR China; Clinical Center for Acute Pancreatitis, Capital Medical University, Beijing, PR China.
Int Immunopharmacol. 2024 Nov 15;141:112923. doi: 10.1016/j.intimp.2024.112923. Epub 2024 Aug 12.
Exogenous inhibition of neutrophil extracellular traps (NETs) was believed to alleviate acute pancreatitis (AP). This study aimed to comprehensively explore the key biological behavior of NETs including timing and pathogenesis in AP by integrating of single cell RNA sequencing(scRNA-seq) and bulk RNA-seq.
Differentially expressed NETs-related genes and the hub genes of NETs were screened by bulk RNA-seq. ScRNA-seq was used to identify the cell types in pancreas of AP mice and to depict the transcriptomic maps in neutrophils. The mouse AP models were build to verify the timing of initiation of NETs and underlying pathogenesis of damage on pancreas acinar cells.
Tlr4 and Ccl3 were screened for hub genes by bulk RNA-seq. The trajectory analysis of neutrophils showed that high expression of Ccl3, Cybb and Padi4 can be observed in the middle stage during AP. Macrophages might be essential in the biological behavior of neutrophils and NETs. Through animal models, we presented that extensive NETs structures were formed at mid-stage of inflammation, accompanied by more serious pancreas and lung damage. NETs might promote necroptosis and macrophage infiltration in AP, and the damage on pancreatic injury could be regulated by Tlr4 pathway. Ccl3 was considered to recruit neutrophils and promote NETs formation.
The findings explored the underlying timing and pathogenesis of NETs in AP for the first time, which provided gene targets for further studies.
细胞外网状陷阱（NETs）的外源抑制被认为可以减轻急性胰腺炎（AP）。本研究旨在通过整合单细胞 RNA 测序（scRNA-seq）和批量 RNA 测序，全面探讨 NETs 的关键生物学行为，包括在 AP 中的时间和发病机制。
通过批量 RNA-seq 筛选差异表达的 NETs 相关基因和 NETs 的枢纽基因。scRNA-seq 用于鉴定 AP 小鼠胰腺中的细胞类型，并描绘中性粒细胞的转录组图谱。构建小鼠 AP 模型以验证 NETs 起始的时间和胰腺腺泡细胞损伤的潜在发病机制。
通过批量 RNA-seq 筛选出 Tlr4 和 Ccl3 作为枢纽基因。中性粒细胞的轨迹分析表明，在 AP 中，Ccl3、Cybb 和 Padi4 的高表达可在中期观察到。巨噬细胞可能在中性粒细胞和 NETs 的生物学行为中至关重要。通过动物模型，我们发现大量 NETs 结构在炎症中期形成，同时胰腺和肺部损伤更严重。NETs 可能在 AP 中促进坏死和巨噬细胞浸润，Tlr4 途径可调节胰腺损伤。Ccl3 被认为可招募中性粒细胞并促进 NETs 形成。
本研究首次探讨了 AP 中 NETs 的潜在时间和发病机制，为进一步研究提供了基因靶点。
