Neuronal injury in rat model of permanent focal cerebral ischemia is associated with activation of autophagic and lysosomal pathways

Autophagy. 2008 Aug;4(6):762-9. doi: 10.4161/auto.6412. Epub 2008 Jun 6.

Abstract

It has been reported that ischemic insult increases the formation of autophagosomes and activates autophagy. However, the role of autophagy in ischemic neuronal damage remains elusive. This study was taken to assess the role of autophagy in ischemic brain damage. Focal cerebral ischemia was introduced by permanent middle cerebral artery occlusion (pMCAO). Activation of autophagy was assessed by morphological and biochemical examinations. To determine the contribution of autophagy/lysosome to ischemic neuronal death, rats were pretreated with a single intracerebral ventricle injection of the autophagy inhibitors 3-methyl-adenine (3-MA) and bafliomycin A1 (BFA) or the cathepsin B inhibitor Z-FA-fmk after pMCAO. The effects of 3-MA and Z-FA-fmk on brain damage, expression of proteins involved in regulation of autophagy and apoptosis were assessed with 2,3,5-triphenyltetrazolium chloride (TTC) staining and immunoblotting. The results showed that pMACO increased the formation of autophagosomes and autolysosomes, the mRNA and protein levels of LC3-II and the protein levels of cathepsin B. 3-MA, BFA and Z-FA-fmk significantly reduced infarct volume, brain edema and motor deficits. The neuroprotective effects of 3-MA and Z-FA-fmk were associated with an inhibition on ischemia-induced upregulation of LC3-II and cathepsin B and a partial reversion of ischemia-induced downregulation of cytoprotective Bcl-2. These results demonstrate that ischemic insult activates autophagy and an autophagic mechanism may contribute to ischemic neuronal injury. Thus, autophagy may be a potential target for developing a novel therapy for stroke.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenine / analogs & derivatives
  • Adenine / metabolism
  • Amino Acid Chloromethyl Ketones / metabolism
  • Animals
  • Autophagy / physiology*
  • Biomarkers / metabolism
  • Brain / metabolism
  • Brain / pathology
  • Brain Ischemia* / metabolism
  • Brain Ischemia* / pathology
  • Cathepsin B / genetics
  • Cathepsin B / metabolism
  • Cysteine Proteinase Inhibitors / metabolism
  • Humans
  • Infarction, Middle Cerebral Artery
  • Lysosomes / metabolism*
  • Male
  • Neurons / cytology
  • Neurons / metabolism*
  • Neurons / pathology*
  • Neuroprotective Agents / metabolism
  • Phagosomes / metabolism
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • Rats
  • Rats, Sprague-Dawley
  • Signal Transduction / physiology

Substances

  • Amino Acid Chloromethyl Ketones
  • Biomarkers
  • Cysteine Proteinase Inhibitors
  • Neuroprotective Agents
  • Proto-Oncogene Proteins c-bcl-2
  • benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone
  • 3-methyladenine
  • Cathepsin B
  • Adenine