Allicin attenuates necrotizing enterocolitis via PINK1/Parkin-mediated mitophagy to suppress pyroptosis

Necrotizing enterocolitis (NEC) is a common and severe gastrointestinal disease in neonates, with unclear pathogenesis and limited therapeutic options. This study explores the intestinal protective effects of allicin in NEC and its potential molecular mechanisms. In lipopolysaccharide (LPS)-induced in vitro NEC model using IEC-6 cells, allicin significantly improved cell viability, reduced cytotoxicity, inhibited apoptosis, promoted proliferation, and suppressed ROS overproduction and inflammatory responses (IL-1β, IL-6, IL-18, TNF-α). Expanding to a physiologically relevant 3D intestinal organoid model mimicking NEC pathology, allicin enhanced organoid budding efficiency and proliferative capacity while restoring epithelial barrier integrity through E-cadherin upregulation, highlighting its ability to repair multicellular structural damage. In a neonatal rat NEC model, allicin treatment reduces mortality in NEC rats, alleviates intestinal pathological damage, and restores intestinal barrier integrity while suppressing intestinal inflammation and pyroptosis, as evidenced by downregulated NLRP3, Cleaved-Caspase-1, and N-terminal-GSDMD expression. Notably, the therapeutic benefits of allicin were largely abolished by the mitophagy inhibitor 3-MA at both cellular and animal levels, underscoring the necessity of mitophagy for its protective effects. Mechanistic studies indicate that allicin activates the PINK1/Parkin-dependent mitophagy pathway, effectively reducing mitochondrial ROS levels and inhibiting pyroptosis. Furthermore, genetic knockdown of PINK1 or Parkin partially reversed allicin-mediated cytoprotection, ROS reduction, and anti-pyroptotic effects, definitively establishing PINK1/Parkin-dependent mitophagy as the crucial pathway. Our findings provide substantial theoretical and scientific support for the potential of allicin as a therapeutic agent for NEC.