Stroke is a leading cause of death and permanent disability in adults worldwide. Intravenous recombinant tissue plasminogen activator (rt- PA) is the only FDA-approved treatment for administration within 3 hours of onset of acute ischemic stroke. The current study has investigated neuroprotective effects of human serum albumin (20% solution) (HSA) in a vehicle (sodium chloride 0.09%) (0.04ml/ 20g) was administrated intravenous at a constant rate over 3 min 2 h after the onset of ischemia. Pioglitazone (40 mg/kg/day, as a suspension in 0.5% carboxymethylcellulose) was orally administrated for three days before ischemia and for two days after ischemia. Insulin was administrated 0.3 IU/kg (via tail vein) at the beginning of MCAO and after 6 h of MCAO. Human serum albumin (HAS), pioglitazone and insulin demonstrated neuroprotective effects against brain damage induced by permanent MCAO in mice as evidenced by the reduction in initiation of walking time and prevention of leukocyte infiltration and brain edema.