Cistanche deserticola exosome-like nanoparticles alleviate constipation through PINK1-dependent mitophagy and gut microbiota remodeling

Introduction Constipation is a prevalent gastrointestinal disorder with limited long-term treatment options. This study investigates the therapeutic potential and mechanisms of Cistanche deserticola exosome-like nanoparticles (CELNs) in alleviating constipation, focusing on mitochondrial quality control and gut microbiota regulation. Methods CELNs were isolated and characterized using ultracentrifugation, transmission electron microscopy, and nanoparticle tracking analysis. Using a loperamide-induced mouse model, constipation parameters, histopathology, inflammation, and gut microbiota were assessed after CELNs treatment. The role of PTEN-induced kinase 1 (PINK1) was investigated with knockout mice, and mechanisms were analyzed via molecular and sequencing approaches. Results CELNs treatment significantly improved defecation, moisture content, and intestinal transit in constipated mice, reduced colonic inflammation, and restored gut morphology. Transcriptomic analysis revealed CELNs enriched pathways related to mitophagy and apoptosis. Crucially, CELNs activated PINK1/Parkin-mediated mitophagy, enhanced mitochondrial function, and inhibited apoptosis of interstitial cells of Cajal. These benefits were abolished in PINK1 −/− mice. Furthermore, CELNs reversed constipation-associated gut microbiota dysbiosis, specifically enriching genera like Alistipes and Bacteroides . This microbial restoration was also PINK1-dependent. Discussion In summary, this study demonstrates that CELNs effectively alleviate constipation through a dual-pathway mechanism centered on PINK1. CELNs activate PINK1/Parkin-mediated mitophagy to clear damaged mitochondria, thereby restoring energy homeostasis and suppressing ICC apoptosis. Simultaneously, CELNs remodel the gut microbiota in a PINK1-dependent manner, which may reinforce mitophagy activation. These findings elucidate a novel exosome-PINK1-mitophagy axis essential for restoring intestinal motility and highlight CELNs as a promising multi-target therapeutic strategy for functional gastrointestinal disorders.