scRNA-seq analysis shows neutrophil extracellular traps as drivers of obstruction-induced intestinal damage in rats

Intestinal obstruction is a common surgical emergency characterised by rapid progression and high mortality. However, the underlying mechanisms remain incompletely elucidated. Here, we show, by combining single-cell RNA sequencing with in vivo pharmacological modulation in male rats, the temporal dynamics of neutrophil extracellular traps (NETs) formation and their contribution to barrier dysfunction in incarcerated small-bowel obstruction. Profiling of 23,320 intestinal cells reveals pronounced neutrophil infiltration in late-stage obstruction (24.59% of total cells) and enables identification of a discrete "NETs-associated neutrophil subset (Neu_NETs)". These Neu_NETs exhibit marked transcriptional enrichment of IL-17, NF-κB and TNF signalling pathways. Functional analyses demonstrate a time-dependent accumulation of NETs within obstructed segments that inversely correlates with the expression of intestinal tight junction proteins. In vivo administration of the PAD4 inhibitor Cl-amidine significantly attenuates NETs formation, ameliorates histopathological injury, and decreases local levels of IL-1β and TNF-α. Therefore, targeting NETs may represent a strategy for treating intestinal obstruction.