title: Mammalian target of rapamycin cell signaling pathway contributes to the protective effects of ischemic postconditioning against stroke.
abstract: OBJECTIVE
  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.affiliation: 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.).
authors.forename: Rong
authors.initials: R
authors.lastname: Xie
authors.affiliation: 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.).
authors.forename: Peng
authors.initials: P
authors.lastname: Wang
authors.affiliation: 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.).
authors.forename: Michelle
authors.initials: M
authors.lastname: Cheng
authors.affiliation: 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.).
authors.forename: Robert
authors.initials: R
authors.lastname: Sapolsky
authors.affiliation: 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.
authors.forename: Xunming
authors.initials: X
authors.lastname: Ji
authors.affiliation: 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.
authors.forename: Heng
authors.initials: H
authors.lastname: Zhao
mesh_terms: D000818:Animals; D001786:Blood Glucose / Q000378:metabolism; D001921:Brain / Q000196:embryology; D002470:Cell Survival; D005947:Glucose / Q000378:metabolism; D000860:Hypoxia / Q000473:pathology; D057775:Ischemic Postconditioning / Q000379:methods*; D016086:Lentivirus / Q000235:genetics; D009474:Neurons / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism; D010766:Phosphorylation; D012313:RNA / Q000378:metabolism; D051381:Rats; D019893:Ribosomal Protein S6 Kinases / Q000378:metabolism; D015398:Signal Transduction; D020123:Sirolimus / Q000494:pharmacology; D020521:Stroke / Q000628:therapy*; D058570:TOR Serine-Threonine Kinases / Q000378:metabolism*
chemical_list: D001786:Blood Glucose; D012313:RNA; C546845:mTOR protein, rat; D019893:Ribosomal Protein S6 Kinases; C526901:Rps6kb1 protein, rat; D058570:TOR Serine-Threonine Kinases; D005947:Glucose; D010100:Oxygen; D020123:Sirolimus
keywords: ischemic postconditioning; stroke