Sphingosine kinase 2-mediated autophagy contributes to preconditioning by isoflurane in mouse cortical neurons
Sphingosine kinase 2 (SPK2) and autophagy are both involved in cerebral preconditioning, but whether preconditioning-induced SPK2 upregulation and autophagy activation are linked mechanistically remains to be elucidated.In this study, we used in vitro and in vivo models to explore the role of SPK2-mediated autophagy in isoflurane (ISO) and hypoxic preconditioning (HP).In primary cultured mouse cortical neurons, both ISO and HP induced autophagy, as evidenced by an elevated LC3Ⅱ/LC3Ⅰ ratio and p62 downregulation.ISO and HP protected against subsequent oxygen glucose deprivation (OGD) or glutamate (Glu) injury, while pretreatment with autophagy inhibitors (3-MA or KU55933) abolished preconditioning-induced tolerance.Pretreatment with SPK2 inhibitors (ABC294640 and SK1-Ⅱ) prevented preconditioning-induced autophagy.ISO also induced autophagy in the cortex of C57 mice as shown by western blots for LC3 and p62, immunoflurescence/immunohistochemistry of LC3 and electron microscopy.ISO induced autophagy in mice lacking the SPK1 isoform (SPKl-/-), but not in SPK2-/-mice.S1P and the S1P receptor agonist FTY720 did not protect against OGD in cultured neurons and did not alter the expression of LC3 and p62, suggesting that SPK2-mediated autopbagy and protections are not S1Pdependent.Beclin1 knockdown abolished preconditioning-induced autophagy activation, but SPK2 inhibitors abolished ISO-induced disruption of the Beclin1/Bcl-2 association.Taken together, these results strongly indicate that autophagy is involved in isoflurane preconditioning both in vivo and in vitro and that SPK2 contributes to preconditioning-induced autophagy, possibly by disrupting the Beclin 1/Bcl-2 interaction.
sphingosine kinase 2 isoflurane preconditioning autophagy cortical neurons coimmunoprecipitation
国内会议
江苏省药理学会青年工作委员会成立大会暨药理学科青年科技创新学术研讨会
苏州
英文
181-181
2013-05-24（万方平台首次上网日期，不代表论文的发表时间）