Sarcodon aspratus fruiting polysaccharides (SAFP) exhibit multiple therapeutic properties. In this study, a type 2 diabetes mellitus (T2DM) mouse model was established using a high-fat diet (HFD) and streptozotocin to evaluate the antidiabetic potential of SAFP. Then the benefits of SAFP on glucolipid metabolism, gut barrier integrity and intestinal microbiota were evaluated. The results indicated that SAFP alleviated disturbances in glycolipid metabolism and insulin resistance through activating Adenosine 5′-monophosphate (AMP)-activated protein kinase (AMPK) signaling pathway. Furthermore, SAFP ameliorated hepatic inflammation and hepatic steatosis, as well as restored dysbiosis in hepatic function. Notably, SAFP enhanced intestinal mucosal architecture and strengthened epithelial barrier functionality through upregulated expression of tight junction components such as Zonula occludens-1(ZO-1), Claudin-1, and Occludin proteins. The 16S rRNA analysis indicated that SAFP has the potential to restore the intestinal microbial barrier in T2DM mice through elevation of short-chain fatty acids (SCFAs) concentrations and regulation of microbial community imbalances. This research offers foundational evidence supporting the utilization of SAFP as an innovative dietary supplement or prospective prebiotic component in functional food formulations targeting diabetes management.