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Aim： To examine the effects of a mixed formulation composed of prostaglandin E1 and lithium （PGEI＋Li mixture） on brain damage after cerebral ischemia. The effects of the mixture on protein expression of heat shock proteins （HSPs）, p53, and Bcl-2 were also determined. Methods： Brain ischemia was induced with a permanent middle cerebral artery occlusion （pMCAO） in rats. Rats were treated with a single intravenous administration of PGE1, lithium or a PGEI＋Li mixture immediately after the ischemic insult. The infarct volume and motor behavior deficits were analyzed 24 h after the ischemic insult. The protein levels of HSPTO, glucose-regulated protein 78 （GRP78）, HSP60, Bcl-2, and p53 in the striatum of the ipsilateral hemisphere were examined using immunoblotting. Results： The mixture （PGE1 22.6 nmol/kg＋Li 0.5 mmol/kg） reduced infarct volume and neurological deficits induced by focal cerebral ischemia. Moreover, the mixture had a greater neuroprotective effect against cerebral ischemia compared with PGE1 or lithium alone. The mixture was effective even if it was administered 3 h after ischemia. PGEI＋Li also significantly upregulated cytoprotective HSPTO GRP78, HSP60, and Bcl-2 protein levels, while decreasing p53 expression. Conclusion： These results demonstrated a PGEI＋Li mixture with a therapeutic window of up to 3 h for clinical treatment of cerebral ischemia. The PGEI＋Li mixture potentially exerts a protective effect after stroke through the induction of HSPs and Bcl-2 proteins.