Sevoflurane (Sev) is a widely used anaesthetic agent in clinical patients. Growing evidences indicated that Sev resulted in cognitive impairment via inducing endoplasmic reticulum (ER) stress mediated neurons apoptosis in vivo. However, the underlying molecular mechanisms have not yet fully understood. In this study, we found that Sev exposure suppresses cell viability, and induces apoptosis by activating caspase-3 apoptotic signaling pathway. Our results further verified that Sevtriggers ER stress via upregulating its markers glucose-regulated protein 78 (GRP78), C/EBP homologous protein (CHOP), caspase-12 and cleaved-PARP proteins. Recently, microRNAs (miRNAs) have been proven to regulate ER stress in a variety of cells, especially neuronal cells. Therefore, we performed the microarray analysis to identified miRNA levels in HT22 cells after treatment with Sev. Our results showed that Sev induces miRNAs aberrant expression and miR-15b-5p was one of the miRNAs being most upregulated in HT22 cells. Furthermore, the Sev-induced apoptosis and ER stress were rescued by knockdown of miR-15b-5p. Additionally, we demonstrated that miR-15b-5p suppresses Rab1A, a regulator in inducing ER stress, by directly targeting its 3’-UTR in HT22 cells. These results suggested that Sev exposure induces ER stress mediated apoptosis in HT22 cells via regulating miR-15b-5p/Rab1A signaling pathway. These data may provide an important therapeutic strategy for fighting against Sev through ER stress mediated neuronal apoptosis in clinical patients.