Yang Yuansu, Zhou Qiang, Liu Shuyu, Wu Bo, Wu Jing, Xu Yangni
Department of Research and Training, Jinling Hospital, Affiliated Hospital of Medical School, Nanjing University, Nanjing, 210000, Jiangsu Province, People's Republic of China.
Department of Clinical Pharmacy, Jinling Hospital, Affiliated Hospital of Medical School, Nanjing University, Nanjing, 210000, Jiangsu Province, People's Republic of China.
Sci Rep. 2026 Jan 24;16(1):6117. doi: 10.1038/s41598-026-36938-9.
This study investigated the role of neutrophils in intestinal ischemia–reperfusion injury (IRI) in mice. We combined single-cell RNA sequencing (scRNA-seq) with in vivo and in vitro functional assays to characterize cellular dynamics. Single-cell RNA sequencing of IRI model tissue revealed a significant increase in neutrophils and inflammatory monocytes, alongside a decrease in T cells, B cells, and NK cells. In vivo neutrophil depletion markedly alleviated intestinal damage, as indicated by reduced serum diamine oxidase (DAO) and IL-6 levels, improved histopathological scores, and preserved Occludin protein integrity. Mechanistically, scRNA-seq identified a pro-inflammatory neutrophil subcluster (C5) characterized by enrichment of endoplasmic reticulum stress (ERS) markers, particularly the transcription factor ATF4. In vitro and in vivo studies confirmed that neutrophils exacerbate IRI severity by inducing ERS via the ATF4 pathway. Pharmacological inhibition of ERS or genetic ablation of ATF4 significantly attenuated neutrophil-driven inflammation and mucosal injury. These findings demonstrate that a specific neutrophil subpopulation aggravates intestinal IRI through the intrinsic ERS/ATF4 pathway, providing a novel perspective on IRI pathophysiology and highlighting a potential therapeutic target for mitigating intestinal damage.
The online version contains supplementary material available at 10.1038/s41598-026-36938-9.
本研究调查了中性粒细胞在小鼠肠道缺血再灌注损伤（IRI）中的作用。我们将单细胞RNA测序（scRNA-seq）与体内和体外功能试验相结合，以表征细胞动态变化。IRI模型组织的单细胞RNA测序显示，中性粒细胞和炎性单核细胞显著增加，同时T细胞、B细胞和NK细胞减少。体内中性粒细胞耗竭显著减轻了肠道损伤，表现为血清二胺氧化酶（DAO）和白细胞介素-6水平降低、组织病理学评分改善以及紧密连接蛋白（Occludin）蛋白完整性得以保留。机制上，scRNA-seq鉴定出一个促炎性中性粒细胞亚群（C5），其特征是内质网应激（ERS）标志物富集，特别是转录因子ATF4。体内外研究证实，中性粒细胞通过ATF4途径诱导ERS，从而加剧IRI的严重程度。对ERS的药理学抑制或ATF4的基因敲除显著减轻了中性粒细胞驱动的炎症和黏膜损伤。这些发现表明，一个特定的中性粒细胞亚群通过内在的ERS/ATF4途径加重肠道IRI，为IRI的病理生理学提供了新的视角，并突出了减轻肠道损伤的潜在治疗靶点。
在线版本包含可在10.1038/s41598-026-36938-9获取的补充材料。
