Remimazolam Inhibits Neuronal Apoptosis in Rat Brain via Slit Guidance Ligand 2/Roundabout 4 Signaling Pathway

Abstract

To investigate the effect of remimazolam on neuronal apoptosis in rat brain through slit guidance ligand 2/roundabout 4 signaling pathway. Primary cortical neurons were obtained from specific pathogen free 3 w old male Wistar rats and sub cultured in cell culture dishes. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay detected cell viability; Hochest/propidium iodide double staining and flow cytometry detected cell apoptosis. Western blot detected apoptosis-related gene expression and slit guidance ligand 2/roundabout 4 signaling pathway-related protein expression. In addition, after slit guidance ligand 2 inhibitor administration, Western blot detected the changes in expression levels of key genes in cells. Relative to control group, cell viability in each remimazolam group presented elevation and apoptosis presented depletion (p<0.05) in a dose-dependent manner. With the increase of remimazolam treatment time, Bcl-2associated X protein, cleaved caspase-3 and cleaved poly adenosine diphosphate-ribose polymerase protein levels showed a gradual decrease and anti-apoptotic protein B-cell lymphoma 2 level showed a gradual increase relative to 0 h (p<0.05). At the same time, with the increase of remimazolam treatment time (0, 3, 6, 12 and 24 h), slit guidance ligand 2 and roundabout 4 protein levels increased continuously relative to 0 h. Additionally, relative to remimazolam group, Bcl-2-associated X protein, cleaved caspase-3 and cleaved poly adenosine diphosphate-ribose polymerase showed a gradual up regulation and B-cell lymphoma 2 showed a gradual down regulation in slit guidance ligand 2 inhibitor and remimazolam co-treatment group. Remimazolam pretreatment may inhibit rat cortical neuronal apoptosis through slit guidance ligand 2/ roundabout 4 signaling pathway and providing new ideas for clinical brain protection.