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 observing 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 significantly 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.
ZHANG Jiale
,
LI Jiaren
,
ZHANG Yaxuan
,
LI Chaoran
,
QIAO Jing
,
YAO Lu
,
NIU Yujie
,
HAO Xiaoqiong
. Effect of SphK2 on BV-2 cell injury in oxygen glucose deprivation/reoxygenation models[J]. Journal of Baotou Medical College, 2024
, 40(9)
: 24
-28
.
DOI: 10.16833/j.cnki.jbmc.2024.09.005
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