Abstract The purpose of this study was to investigate the protective effects of sulfated chitosan of C3 sulfation (TCTS) on the glycerol-induced acute renal failure. Compared with the normal group, rats from model group exhibited collecting duct and medullary ascending limb dilation and casts by glycerol treating. TCTS, which was injected to pretreat rats by glycerol, exerted a protective effect. The results showed that serum creatinine and blood urea nitrogen were markedly increased in glycerol-treated rats. It is proved that TCTS reduced their levels significantly. Ions level in plasma and urine were significantly changed in glycerol-treated rats, whereas TCTS almost recovered their levels back to normal. For female rats, administration of TCTS reduced their mortality. This study showed a noticeable renal morphologic and functional protection by TCTS in glycerol-induced acute renal failure.