: To investigate the protective effect and possible mechanism of epidemical growth factor receptor (EGFR) inhibitor PD168393 on the blood spinal cord barrier (BSCB) damage induced by oxygen glucose deprivation/re-oxygenation (OGD/R) in vitro. Methods: Primary microvascular endothelial cells and mixed glial cells from the rat spinal cord were cultured using the transwell culture system to establish the in vitro BSCB model. The cultured cells were divided into three groups: control group (untreated), injury group (BSCB damage induced by OGD/R), and treatment group (10 nM PD168393 intervention after OGD/R). The permeability of endothelial cells in the injury and treatment groups was evaluated by the fluorescein leakage test and transepithelial electrical resistance (TEER) at 3 h, 6 h, and 12 h after reoxygenation. The expression of tight junction proteins ZO-1 and Occludin in the endothelial cells of each group was determined by immunofluorescence and Western blot. The differences in proinflammatory factors interleukin-6 (IL-6), tissue necrosis factor-α (TNF-α), and inducible nitric oxide synthase (iNOS) secreted by each group of cells were measured by ELISA. Results: The fluorescein leakage test showed that the amount of fluorescein leakage in the injury group was higher than that in the control group at every time point (all P<0.05), while the amount of fluorescein leakage in the treatment group was significantly lower than that in the injury group (all P<0.05). TEER value of the injury group was significantly lower than that of the control group (all P<0.01), while TEER value of the treatment group was significantly higher than that of the injury group (all P<0.05). The expression of ZO-1 and Occludin in the injury group was significantly lower than those in the control group (both P<0.05), and the expression of these proteins in the treatment group was significantly higher than those in the injury group (both P<0.05). The expression levels of IL-6, TNF-α, and iNOS in the injury group were significantly higher than those in the control group at every time point, and the levels in the treatment group were lower than those in the injury group at every time point (all P<0.05). Conclusion: The EGFR inhibitor PD168393 can preserve the integrity of the BSCB after OGD/R. The mechanism may involve reducing the expression of proinflammatory factors and suppressing the destruction of tight junction proteins in the BSCB.