Research ReportRegulation on Beclin-1 expression by mTOR in CoCl2-induced HT22 cell ischemia-reperfusion injury

- Beclin-1 and p-mTOR were induced by CoCl2-induced I/R injury in HT22 cells.
- The dynamic change of Beclin-1 and p-mTOR in CoCl2-induced I/R injury was observed.
- Beclin-1 and mTOR may have coordinated regulation in ischemia but not in reperfusion.
- Rapamycin could affect the level of Beclin-1 in I/R injury.
- Rapamycin could accelerate cell death in ischemic, but protect the cells against reperfusion injury.

It has been reported that cerebral ischemia/reperfusion (I/R) injury can activate autophagy. However, the role of autophagy in cerebral I/R injury remains controversy. Two major proteins, mTOR and Beclin-1, govern the formation of autophagosomes to regulate autophagy activity. However, the cross-talking between Beclin-1 and mTOR in cerebral I/R injury remains elusive. In this study, global cerebral I/R injury animal model and focal cerebral I/R injury animal model were induced to test the variation of Beclin-1 level in vivo. To further confirm the variation of Beclin-1 level and investigate the cross-talking between Beclin-1 and mammalian target of rapamycin (mTOR) in I/R injury, we used cobalt chloride (CoCl2) to develop an I/R injury cell model in HT22 cell line. Our data showed that the levels of Beclin-1 and phosphorylated mammalian target of rapamycin (p-mTOR) were clearly induced by I/R injury in vitro. And the time course studies suggested that the Beclin-1 and mTOR may have coordinated regulation in ischemia stages but not in reperfusion stages. Moreover, inhibitor of mTOR could prevent Beclin-1 decreasing, but this prevention may play opposite roles in different stages of I/R injury. We conclude that this study represents a major advance in our understanding of the cross-talking of two key proteins, Beclin-1 and mTOR, in autophagy and the role of autophagy in cerebral I/R injury.