Wheat Peptides Promote Recovery of the Enteric Nervous System via mAChR Activation to Relieve Constipation in Mice

Constipation represents a prevalent gastrointestinal disorder. This study demonstrates that wheat peptides (WP) effectively ameliorate loperamide-induced constipation in mice. Administration of WP orally at 0.5 mg/g body weight led to significant improvements in defecation, small intestinal motility, and the integrity of the intestinal barrier. WP further modulated inflammatory markers and enteric nervous system (ENS) regulators while correcting gut microbiota dysbiosis through restoration of microbial diversity and taxonomic composition. Crucially, colon transcriptomics revealed WP-mediated activation of calcium signaling pathway genes and the acetylcholine receptor gene. RT-qPCR validation confirmed the upregulation of key effectors, including muscarinic acetylcholine receptor Chrm1, inositol trisphosphate receptors (Itpr2/3), and calmodulin-like proteins (Calml3, Calm4). Three-dimensional imaging substantiated WP-induced augmentation of enteric neurons in constipated colons. Collectively, these results indicate that WP alleviates constipation through a multitarget mechanism involving muscarinic receptor activation, potentiation of calcium-dependent signaling, and promotion of enteric nervous system recovery, which collectively restore normal intestinal motility. WP thus emerges as a promising therapeutic candidate for constipation management.