This study investigated the role of glycogen synthase kinase-3 beta (GSK-3 beta) in isoflurane-induced neuroinflammation and cognitive dysfunction in aged rats. The hippocampi were dissected from aged rats which had been intraperitoneally administered lithium chloride (LiCl, 100 mg/kg) and then exposed to 1.4% isoflurane for 6 h. The expression of GSK-3 beta was detected by Western blotting. The mRNA and protein expression levels of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1 beta and IL-6 were measured by real-time PCR and enzyme-linked immunosorbent assay (ELISA), respectively. Morris water maze was employed to detect spatial memory ability of rats. The results revealed that the level of GSK-3 beta was upregulated after isofurane exposure. Real-time PCR analysis demonstrated that isoflurane anesthesia increased mRNA levels of TNF-alpha, IL-1 beta and IL-6, which was consistent with the ELISA results. However, these changes were reversed by prophylactic LiCl, a non-selective inhibitor of GSK-3 beta. Additionally, we discovered that LiCl alleviated isoflurane-induced cognitive impairment in aged rats. Furthermore, the role of GSK-3 beta in isoflurae-induced neuroinflammation and cognitive dysfunction was associated with acetylation of NF-kappa B p65 (Lys310). In conclusion, these results suggested that GSK-3 beta is associated with isoflurane-induced upregulation of proinflammatory cytokines and cognitive disorder in aged rats.