AIM: To examine the effects of three commonly used general anesthetics on the proliferation and activation of glial cells in neonatal rats. METHODS: Neonatal rats were exposed to either isoflurane, sevoflurane or desflurane for 2 h on postnatal day 2 (P2). The animals were euthanatihed and the brain were harvested on P7 and P14, respectively. The immunohistochemical localihation of glial markers (vimentin, GFAP, Iba1) were examined. RESULTS: Activation of astrocyte in granular layer and molecular layer of dentate gyrus of hippocampus was significantly enhanced on P7 and P14 after desflurane exposure, while that in isoflurane group the change was only significantly different on P14. The activation of microglia in the granular layer of dentate gyrus but not in the pyramidal cell layer of CA1 region was significantly enhanced in the desflurane group on P7 and P14, while the isoflurane group only showed significant difference on P14. CONCLUSION: Short time exposure of different inhalation anesthetics has different effects on the activation of glial cells in different subregions of hippocampus in neonatal rats on postnatal day 2, and sevoflurane may have the least effect on it.