Endoplasmic reticulum stress (ERS) is associated with many nervous system diseases. IRElα is considered as ERS sensor that, upon activation, initiates the nonconventional splicing of the precursor unspliced form of X-box binding protein 1 (XBP1u) messenger RNA (mRNA) to yield an active transcription factor-XBP1s. The goal of this study is to detect whether there is activation of IRE1α-XBP1 pathway in the medial prefrontal cortex (mPFC) of posttraumatic stress disorder (PTSD) model rats. This study adopted single-prolonged stress (SPS) model. Behavioral functions including anxiety-like behavior, exploration behavior, and spatial memory were assessed by open field test and Morris water maze test. We detected the IRE1α and XBP1 by using methods of double-labeling immunofluorescence, Western blot, and quantitative real-time reverse transcription-PCR (qRT-PCR). We also observed neuronal apoptosis by transferase-mediated dUTP Nick-end-labeling (TUNEL) staining and the expression of caspase-12 by qRT-PCR. Our results showed that the expression of IRE1α, XBP1u, and total XBP1 significantly increased at 1 day after SPS and then decreased gradually. At the same time, XBP1s appeared and peaked at 4 days after SPS, which indicated that IRE1α-XBP1 pathway was activated upon induction of SPS stimulation. We also noted that the mRNA of caspase-12 was upregulated after SPS. Our study preliminarily showed that ERS mediated by IRE1α-XBP1 pathway was closely related to PTSD and it might be a pathogenesis of PTSD.