miR-329-3p derived from mast cells-exosomes exacerbates intestinal ischemia-reperfusion injury by targeting ATG10 to regulate autophagy.

Abstract

Activation and degranulation of intestinal mucosal mast cells (IMMCs) play a critical role in intestinal ischemia-reperfusion (IIR) injury. Exosomes can regulate various physiological and pathological processes by mediating intercellular communication. However, whether activated mast cells modulate IIR injury via exosome secretion remains unclear. In this study, exosomes were isolated from IMMCs of sham-operated and IIR model mice, followed by microRNA (miRNA) high-throughput sequencing to identify significantly altered miRNA, particularly miR-329-3p. First, the impact of oxygen-glucose deprivation/reperfusion (OGD/R)-treated human mast cell (HMC-1)-derived exosomes (OGD/R-HMC-exo) on OGD/R-induced injury in Caco-2 cells was investigated in vitro. Subsequently, the role of miR-329-3p in IIR injury via targeting ATG10 was explored through bioinformatics analysis, cellular experiments, and animal studies. Mechanistic investigations were performed using 5-ethynyl-2-deoxyuridine (EdU) assay, Western blot, reverse transcription quantitative polymerase chain reaction (RT-qPCR), small interfering RNA (siRNA), dual-luciferase reporter assay, and flow cytometry. The research results show that exosomal OGD/R inhibited autophagy, thereby promoting apoptosis and ultimately exacerbating OGD/R injury in Caco-2 cells. After IIR injury, miR-329-3p levels were significantly elevated in exosomes secreted by IMMCs. Upregulation of miR-329-3p downregulated ATG10 expression, suppressed autophagy, impaired autophagic flux, and subsequently aggravated apoptosis, compromised intestinal mucosal barrier integrity, and ultimately exacerbated IIR injury as well as OGD/R injury in Caco-2 cells. These findings provide a novel potential therapeutic target and theoretical basis for the prevention and treatment of IIR injury.