A Neutrophil–Epithelial Thbs1-CD36-JNK Axis Orchestrates Mucosal Barrier Failure in Intestinal Ischemia-Reperfusion

Background Intestinal barrier failure following ischemia–reperfusion (IR) injury is a critical event that drives lethal systemic complications; however, the specific intercellular signals governing epithelial cell fate under these pathological conditions remain incompletely understood. Methods We performed single-cell RNA sequencing (scRNA-seq) on intestinal tissues from a murine IR model to map intercellular communication networks. The identified ligand–receptor pair was functionally validated using neutrophil-specific thrombospondin-1 (Thbs1) knockout mice, combined with pharmacological inhibition and activation of CD36, as well as biochemical and histological analyses. Results scRNA-seq analysis of the IR-injured gut revealed a key neutrophil–epithelial signaling pathway mediated by the Thbs1–CD36 ligand–receptor pair. Neutrophil-specific genetic ablation of Thbs1 markedly attenuated IR-induced epithelial apoptosis and barrier disruption. Mechanistically, Thbs1 binding to epithelial CD36 activated JNK signaling, mediating a proapoptotic cascade. CD36 inhibition was protective, whereas its activation bypassed the requirement for Thbs1 and provoked mucosal injury. Conclusions Our study elucidates a key mechanism of gut barrier failure and identifies the Thbs1–CD36–JNK axis as a promising therapeutic target for mitigating intestinal IR injury in critical conditions such as burns and shock.