The present work was performed to evaluate the effect of acidic-extractable polysaccharides (AE-PS) from fruit bodies of Cordyceps militaris on type 2 diabetes mellitus (T2DM) and its structural characteristics. The T2DM mice induced by high-fat diet (HFD) and streptozotocin (STZ) were administered with 100 and 400 mg/kg AE-PS for 4 weeks. Our work proved that AE-PS decreased the levels of serum lipid, lipid peroxidation, and blood glucose; improved glucose and insulin resistance; enhanced antioxidant enzyme activities; and attenuated the injuries of the liver, kidney, and pancreas in T2DM mice. These results might offer references for the exploitation of AE-PS as functional foods or natural drug source for preventing and treating HFD- and STZ-induced T2DM. Moreover, gas chromatography (GC) results revealed that AE-PS was heterogeneous and composed of fucose, ribose, arabinose, xylose, mannose, galactose, and glucose with mass percentages of 1.23%, 0.57%, 0.29%, 2.12%, 2.73%, 4.66%, and 88.4%, respectively. Fourier-transform infrared (FTIR) and nuclear magnetic resonance (NMR) analysis indicated that AE-PS was a pyran-type polysaccharide with α- and β-configurations.