title: GSK-3β inhibitor TDZD-8 reduces neonatal hypoxic-ischemic brain injury in mice.
abstract: OBJECTIVE
  Glycogen synthase kinase 3β (GSK-3β) is activated following hypoxic-ischemic (HI) brain injury. TDZD-8 is a specific GSK-3β inhibitor. Currently, the impact of inhibiting GSK-3β in neonatal HI injury is unknown. We aimed to investigate the effect of TDZD-8 following neonatal HI brain injury.
  
  
  METHODS
  Unilateral common carotid artery ligation followed by hypoxia was used to induce HI injury in postnatal day 7 mouse pups pretreated with TDZD-8 or vehicle. The infarct volume, whole-brain imaging, Nissl staining, and behavioral tests were used to evaluate the protective effect of TDZD-8 on the neonatal brain and assess functional recovery after injury. Western blot was used to evaluate protein levels of phosphorylated protein kinase B (Akt), GSK-3β, and cleaved caspase-3. Protein levels of cleaved caspase-3, neuronal marker, and glial fibrillary acidic protein were detected through immunohistochemistry.
  
  
  RESULTS
  Pretreatment with TDZD-8 significantly reduced brain damage and improved neurobehavioral outcomes following HI injury. TDZD-8 reversed the reduction of phosphorylated Akt and GSK-3β, and the activation of caspase-3 induced by hypoxia-ischemia. In addition, TDZD-8 suppressed apoptotic cell death and reduced reactive astrogliosis.
  
  
  CONCLUSIONS
  TDZD-8 has the therapeutic potential for hypoxic-ischemic brain injury in neonates. The neuroprotective effect of TDZD-8 appears to be mediated through its antiapoptotic activity and by reducing astrogliosis.
authors.affiliation: Department of Physiology, University of Toronto, Toronto, ON, Canada.
authors.forename: Sammen
authors.initials: S
authors.lastname: Huang
authors.affiliation: Department of Physiology, University of Toronto, Toronto, ON, Canada.
authors.forename: Haitao
authors.initials: H
authors.lastname: Wang
authors.affiliation: Department of Physiology, University of Toronto, Toronto, ON, Canada.
authors.forename: Ekaterina
authors.initials: E
authors.lastname: Turlova
authors.affiliation: Department of Physiology, University of Toronto, Toronto, ON, Canada.|Department of Surgery, University of Toronto, Toronto, ON, Canada.
authors.forename: Ahmed
authors.initials: A
authors.lastname: Abussaud
authors.affiliation: Department of Physiology, University of Toronto, Toronto, ON, Canada.|Department of Surgery, University of Toronto, Toronto, ON, Canada.
authors.forename: Xiang
authors.initials: X
authors.lastname: Ji
authors.affiliation: Department of Physiology and Biophysics, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil.
authors.forename: Luiz R
authors.initials: LR
authors.lastname: Britto
authors.affiliation: Department of Paediatrics, Hospital for Sick Children and University of Toronto, Toronto, ON, Canada.
authors.forename: Steven P
authors.initials: SP
authors.lastname: Miller
authors.affiliation: Centro de Investigaciones Biologicas-CSIC, Madrid, Spain.
authors.forename: Ana
authors.initials: A
authors.lastname: Martinez
authors.affiliation: Department of Physiology, University of Toronto, Toronto, ON, Canada.|Department of Surgery, University of Toronto, Toronto, ON, Canada.
authors.forename: Hong-Shuo
authors.identifier: http://orcid.org/0000-0001-5142-1874
authors.initials: HS
authors.lastname: Sun
authors.affiliation: Department of Physiology, University of Toronto, Toronto, ON, Canada.
authors.forename: Zhong-Ping
authors.initials: ZP
authors.lastname: Feng
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D001253:Astrocytes / Q000187:drug effects / Q000201:enzymology / Q000473:pathology; D001921:Brain / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D053148:Caspase 3 / Q000378:metabolism; D004268:DNA-Binding Proteins; D004195:Disease Models, Animal; D004353:Drug Evaluation, Preclinical; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D005911:Gliosis / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology; D000071679:Glycogen Synthase Kinase 3 beta / Q000037:antagonists & inhibitors* / Q000378:metabolism; D020925:Hypoxia-Ischemia, Brain / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D051379:Mice; D009043:Motor Activity / Q000187:drug effects / Q000502:physiology; D009419:Nerve Tissue Proteins / Q000378:metabolism; D018696:Neuroprotective Agents / Q000494:pharmacology*; D009687:Nuclear Proteins / Q000378:metabolism; D010766:Phosphorylation; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D011897:Random Allocation; D013830:Thiadiazoles / Q000494:pharmacology*
chemical_list: C494356:4-benzyl-2-methyl-1,2,4-thiadiazolidine-3,5-dione; D004268:DNA-Binding Proteins; D005904:Glial Fibrillary Acidic Protein; D009419:Nerve Tissue Proteins; C492380:NeuN protein, mouse; D018696:Neuroprotective Agents; D009687:Nuclear Proteins; D047428:Protein Kinase Inhibitors; D013830:Thiadiazoles; C528900:glial fibrillary astrocytic protein, mouse; D000071679:Glycogen Synthase Kinase 3 beta; C000605507:Gsk3b protein, mouse; D051057:Proto-Oncogene Proteins c-akt; C505409:Casp3 protein, mouse; D053148:Caspase 3
keywords: GSK-3β; TDZD-8; neonatal hypoxia-ischemia, neuroprotection