Effect of SphK2 on BV-2 cell injury in oxygen glucose deprivation/reoxygenation models
Objective:To investigate the expression of sphingosine kinase 2(SphK2)in BV-2 microglia during oxygen glucose deprivation/reoxygenation(OGD/R),and to investigate whether SphK2 is involved in the damage process of BV-2 microglia induced by OGD/R.Methods:The OGD/R model was established using mouse BV-2 microglial cells.The oxygen glucose deprivation time was 2 hours,and the reoxygenation and rehydration time was 0,3,6,12,and 24 hours.(1)The control group,OGD2h group,OGD2h/R3h group,OGD2h/R6h group,OGD2h/R12h group and OGD2h/R24h group were set up to observe the changes of cell morphology.Cell viability was detected by CCK-8 method after OGD/R treatment;(2)The control group,OGD2h/R12h group and OGD2h/R12h+ABC294640(SphK2 specific inhibitor)group were set up,and the expression of SphK2 gene and protein was determined by q-PCR and Western blot.Results:Compared with the control group,with the prolongation of OGD/R time,the activity of BV-2 gradually decreased(P＜0.001),and the cell morphology changed to varying degrees,with varying degrees of injury By ob-serving cell survival,OGD2h/R12h was determined as the most suitable OGD/R time.Compared with the control group,the expression level of Sphk2 mRNA in BV-2 cells treated with OGD/R was signif-icantly increased(P＜0.05),and the addition of ABC294640 significantly inhibited the increase in its expression(P＜0.001).Compared with the control group,the expression level of SphK2 protein in BV-2 cells treated with OGD/R was significantly increased(P＜0.001),while the expression of SphK2 protein was significantly decreased after adding ABC294640(P＜0.001).Conclusion:SphK2 activates microglia after OGD/R injury and plays a regulatory role in the process of OGD/R injury.
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