A neutrophil–epithelial thrombospondin-1–CD36–c-Jun N-terminal kinase axis orchestrates mucosal barrier failure in intestinal ischemia–reperfusion

Abstract

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 ribonucleic acid 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.

Keywords

Intestinal ischemia–reperfusion injury / Neutrophils / Apoptosis / Thrombospondin-1 / CD36 / JNK signaling / Intestinal mucosal barrier

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