Three extractable polysaccharides including Ac-RPS, Al-RPS and En-RPS were extracted from the residue of Flammulina velutipes and their antioxidative and renoprotective effects on STZ-induced mice were investigated. Biochemical and antioxidant analysis showed that the En-RPS had potential effects in decreasing the serum levels of CRE, BUN, ALB and GLU significantly, increasing the renal activities of SOD, CAT and GSH-Px remarkably, and reducing the renal contents of MDA prominently. Furthermore, the histopathological observations also displayed that En-RPS could alleviate kidney damage. These results demonstrated that En-RPS extracted from the residue of F. velutipes possessed potent antioxidant activities, and could be used as a promising therapeutic agent for inhibiting the progression of diabetic nephropathy. In addition, the monosaccharide compositions of these three RPS were also analyzed.