Rong Xie1, Peng Wang1, Michelle Cheng1, Robert Sapolsky1, Xunming Ji2, Heng Zhao2. 1. From the Departments of Neurosurgery (R.X., P.W., M.C., R.S., H.Z.) and Biological Sciences (M.C., R.S.), and Stanford Stroke Center (H.Z.), Stanford University, CA; Department of Neurosurgery, Huashan Hospital, Fudan University, Shanghai, China (R.X.); and Department of Neurosurgery, Xuanwu Hospital, Capital Medical School, Beijing, China (P.W., X.J.). 2. From the Departments of Neurosurgery (R.X., P.W., M.C., R.S., H.Z.) and Biological Sciences (M.C., R.S.), and Stanford Stroke Center (H.Z.), Stanford University, CA; Department of Neurosurgery, Huashan Hospital, Fudan University, Shanghai, China (R.X.); and Department of Neurosurgery, Xuanwu Hospital, Capital Medical School, Beijing, China (P.W., X.J.). hzhao@stanford.edu jixm70@hotmail.com.
Abstract
BACKGROUND AND PURPOSE: Whether the mammalian target of rapamycin (mTOR) pathway is protective against brain injury from stroke or is detrimental is controversial, and whether it is involved in the protective effects of ischemic postconditioning (IPC) against stroke is unreported. Our study focuses on the protective role of mTOR against neuronal injury after stroke with and without IPC. METHODS: We used both an in vitro oxygen-glucose deprivation model with a mixed neuronal culture and hypoxic postconditioning, as well as an in vivo stroke model with IPC. Rapamycin, a specific pharmacological inhibitor of mTOR, and mTOR short hairpin RNA lentiviral vectors were used to inhibit mTOR activity. A lentiviral vector expressing S6K1, a downstream molecule of mTOR, was used to confirm the protective effects of mTOR. Infarct sizes were measured and protein levels were examined by Western blot. RESULTS: We report that stroke resulted in reduced levels of phosphorylated proteins in the mTOR pathway, including S6K1, S6, and 4EBP1, and that IPC increased these proteins. mTOR inhibition, both by the mTOR inhibitor rapamycin and by mTOR short hairpin RNA, worsened ischemic outcomes in vitro and in vivo and abolished the protective effects of hypoxic postconditioning and IPC on neuronal death in vitro and brain injury size in vivo. Overexpression of S6K1 mediated by lentiviral vectors significantly attenuated brain infarction. CONCLUSIONS: mTOR plays a crucial protective role in brain damage after stroke and contributes to the protective effects of IPC.
BACKGROUND AND PURPOSE: Whether the mammalian target of rapamycin (mTOR) pathway is protective against brain injury from stroke or is detrimental is controversial, and whether it is involved in the protective effects of ischemic postconditioning (IPC) against stroke is unreported. Our study focuses on the protective role of mTOR against neuronal injury after stroke with and without IPC. METHODS: We used both an in vitro oxygen-glucose deprivation model with a mixed neuronal culture and hypoxic postconditioning, as well as an in vivo stroke model with IPC. Rapamycin, a specific pharmacological inhibitor of mTOR, and mTOR short hairpin RNA lentiviral vectors were used to inhibit mTOR activity. A lentiviral vector expressing S6K1, a downstream molecule of mTOR, was used to confirm the protective effects of mTOR. Infarct sizes were measured and protein levels were examined by Western blot. RESULTS: We report that stroke resulted in reduced levels of phosphorylated proteins in the mTOR pathway, including S6K1, S6, and 4EBP1, and that IPC increased these proteins. mTOR inhibition, both by the mTOR inhibitor rapamycin and by mTOR short hairpin RNA, worsened ischemic outcomes in vitro and in vivo and abolished the protective effects of hypoxic postconditioning and IPC on neuronal death in vitro and brain injury size in vivo. Overexpression of S6K1 mediated by lentiviral vectors significantly attenuated brain infarction. CONCLUSIONS:mTOR plays a crucial protective role in brain damage after stroke and contributes to the protective effects of IPC.
Authors: Giuseppe Pignataro; Robert Meller; Koichi Inoue; Andrea N Ordonez; Michelle D Ashley; Zhigang Xiong; Rosaria Gala; Roger P Simon Journal: J Cereb Blood Flow Metab Date: 2007-09-19 Impact factor: 6.200