Abstract
INTRODUCTION: This study aimed to systematically fractionate Coprinus comatus mycelial polysaccharides using ethanol precipitation, screen the optimal fraction, and investigate its potential to alleviate inflammation in a dextran sulfate sodium (DSS)-induced Caco-2 cell model. METHODS: Polysaccharides (CCPJ-40, -60, -80) were obtained with 40%, 60%, and 80% ethanol. Their chemical properties (yield, monosaccharide composition, molecular weight) and in vitro bioactivities were assessed. The optimal fraction was investigated in a dextran sulfate sodium (DSS)-induced Caco-2 cell model. RESULTS: The results revealed that the ethanol concentration significantly affected the polysaccharide yield, chemical composition, and bioactivities. Notably, CCPJ-80 exhibited the highest yield and total sugar content. Monosaccharide composition analysis indicated that both CCPJ-40 and CCPJ-80 were composed of L-fucose, L-arabinose, galactose, glucose, xylose, mannose, ribose, and D-glucuronic acid, whereas CCPJ-60 additionally contained rhamnose. Regarding molecular weight distribution, CCPJ-40 had a high molecular weight and was relatively polydisperse, while CCPJ-60 and CCPJ-80 had lower molecular weights with good homogeneity. In the in vitro activity assessment, CCPJ-80 demonstrated the most potent antioxidant and prebiotic activities. Further cell experiments confirmed that CCPJ-80 effectively alleviated the DSS-induced inflammatory state in Caco-2 cells. The underlying mechanisms included significantly reducing the levels of lactate dehydrogenase, tumor necrosis factor-α, interleukin-6, reactive oxygen species, and mitochondrial ROS, while simultaneously enhancing superoxide dismutase activity and restoring the mitochondrial membrane potential. DISCUSSION: This study elucidates the regulatory effect of ethanol precipitation concentration on the physicochemical properties and functional activities of CC mycelial polysaccharides, and reveals the potential of CCPJ-80 to ameliorate ulcerative colitis by modulating oxidative stress and mitochondrial function pathways, providing a scientific basis for developing CCPJ as a functional food or therapeutic agent.