title: PEP-1-metallothionein-III protein ameliorates the oxidative stress-induced neuronal cell death and brain ischemic insults.
abstract: BACKGROUND
  Oxidative stress is considered to be involved in a number of human diseases including ischemia. Metallothioneins (MT)-III can protect neuronal cells from the cytotoxicity of reactive oxygen species (ROS). However, MT-III proteins biological function is unclear in ischemia. Thus, we examined the protective effects of MT-III proteins on oxidative stress-induced neuronal cell death and brain ischemic insult.
  
  
  METHODS
  A human MT-III gene was fused with a protein transduction domain, PEP-1 peptide, to construct a cell permeable PEP-1-MT-III protein. PEP-1-MT-III protein was purified using affinity chromatograph. Transduced PEP-1-MT-III proteins were detected by Western blotting and immunoflourescence. Cell viability and DNA fragmentation were analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-dipheyltetrazolium bromide (MTT) assay and terminal dexoynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining, respectively. Brain ischemic injury was detected with immunohistochemistry.
  
  
  RESULTS
  Purified PEP-1-MT-III proteins transduced into astrocytes in a time- and dose-dependent manner and protected against oxidative stress-induced cell death. Also, transduced PEP-1-MT-III proteins efficiently protected cells against DNA fragmentation. Furthermore, immunohistochemical analysis revealed that PEP-1-MT-III prevented neuronal cell death in the CA1 region of the hippocampus induced by transient forebrain ischemia. We demonstrated that transduced PEP-1-MT-III protein protects against oxidative stress induced cell death in vitro and in vivo.
  
  
  CONCLUSIONS
  Transduced PEP-1-MT-III protein has neuroprotective roles as an antioxidant in vitro and in vivo. PEP-1-MT-III protein is a potential therapeutic agent for various human brain diseases such as stroke, Alzheimer's disease, and Parkinson's disease.
authors: Department of Biomedical Science and Research Institute of Bioscience and Biotechnology, Hallym University, Chunchon 200-702, Korea.
authors: Eun Jeong
authors: EJ
authors: Sohn
authors: Dae Won
authors: DW
authors: Kim
authors: Mi Jin
authors: MJ
authors: Kim
authors: Hoon Jae
authors: HJ
authors: Jeong
authors: Min Jea
authors: MJ
authors: Shin
authors: Eun Hee
authors: EH
authors: Ahn
authors: Soon Won
authors: SW
authors: Kwon
authors: Young Nam
authors: YN
authors: Kim
authors: Duk-Soo
authors: DS
authors: Kim
authors: Kyu Hyung
authors: KH
authors: Han
authors: Jinseu
authors: J
authors: Park
authors: Hyun Sook
authors: HS
authors: Hwang
authors: Won Sik
authors: WS
authors: Eum
authors: Soo Young
authors: SY
authors: Choi
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D002545:Brain Ischemia / Q000188:drug therapy* / Q000473:pathology / Q000517:prevention & control; D016923:Cell Death / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D002478:Cells, Cultured; D003543:Cysteamine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology / Q000627:therapeutic use; D005849:Gerbillinae; D015227:Lipid Peroxidation / Q000187:drug effects / Q000235:genetics; D000090023:Metallothionein 3; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology* / Q000627:therapeutic use; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology; D018384:Oxidative Stress / Q000187:drug effects* / Q000235:genetics; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology* / Q000627:therapeutic use; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000494:pharmacology / Q000627:therapeutic use; D014161:Transduction, Genetic / Q000379:methods
chemical_list: D000090023:Metallothionein 3; D009419:Nerve Tissue Proteins; C503924:Pep-1 peptide; D010455:Peptides; D011993:Recombinant Fusion Proteins; D003543:Cysteamine