Effect Of S1P1 Receptor Pathway Inhibition On Astrocyte Precursor Cell Proliferation Following Oxygen-glucose Deprivation In Vitro

Objective:To observe the influence on astrocytes of proliferation after the Oxygen-glucose deprivation treated with W146 which is the antagonist of the sphingosine 1-phosphate receptor type 1. Methods:The cerebral cortex tissue of SD neonatal rat within three days was used to culture primary astrocytes. The third generation cell would be identified its purity by immunofluoresence. The immunofluresence would be used to identified the protein expression of S1P1 R on astrocytes. Cell inoculated in Petri dishes was divided into three groups randomly: CONTROL Group, OGD/R Group and OGD/R+W146 Group. OGD mimic the hypoxia injury in vitro. Experiment with MTT to detect the proliferative activity of astroglia 12 hour, 24 hour, 48 hour and 72 hour after the reperfusion. Use the Edu and flow cytometry to detect the cell fraction under the proliferative stage(phase S). Western blot would be used to detect the protein level of the S1P1 R expressed in the astrocytes. Result:Compared with OGD/R Group, the multiplication capacity of the cells in OGD+W146 Group has declined(P<0.05). Compared with ODG/R Group, the cell fraction of the OGD/R+W146 Group under the proliferative stage has decline obviously(6 hour and 12 hour, P<0.05). At the same time, the expression level of S1P1 R of OGD/R+W146 Group has declined when compared with OGD/R Group(6 hour and 12 hour, P<0.05). Conclusions:W146 can inhibit proliferation of astrocytes after OGD.