title: Long-term effects of hypothermia on neuronal cell death and the concentration of apoptotic proteins after incomplete cerebral ischemia and reperfusion in rats.
abstract: BACKGROUND
  The present study investigates the long-term effects of postischemic hypothermia on neuronal cell damage and concentration changes of apoptotic proteins after cerebral ischemia.
  
  
  METHODS
  Sixty-four Sprague-Dawley rats were anesthetized, intubated and ventilated with 2.0 Vol% isoflurane and 70% N2O/O2. After preparation the animals were randomly assigned to the following groups: group 1 (n = 32, fentanyl-N2O/normothermia 37.5 degrees C), and group 2 (n = 32, fentanyl-N2O/hypothermia 34.0 degrees C. Ischemia (45 min) was induced by common carotid artery occlusion plus hemorrhagic hypotension (MAP = 40 mmHg). Arterial blood gases and pH were maintained constant. After 1, 3, 7, or 28 days (each n = 8) the brains were removed, frozen and cut. Neuronal damage was assessed by analyzing Bax, Bcl-2, p53, and Mdm-2 proteins, activated caspases-3-positive and eosinophilic cells. A third group (n = 8) of untreated animals served as naive controls.
  
  
  RESULTS
  In hypothermic animals, Bax concentration was decreased by 50-70% over time compared to normothermia. On days 1 and 3, Bcl-2 was increased by 50% with hypothermia. The amount of activated caspase-3-positive cells in the ischemic hemisphere was 0.5% in the hypothermic and 1-2% in the normothermic animals. Of the hippocampal cells, 10-25% were eosinophilic in both groups over time.
  
  
  CONCLUSIONS
  The present data show that hypothermia prevents an ischemia-induced increase of the pro-apoptotic protein Bax for as long as 28 days and increases the concentration of the antiapoptotic protein Bcl-2 up to 3 days compared to normothermic animals. Therefore, after cerebral ischemia, hypothermia has the sustained neuroprotective potential to shift apoptosis-related proteins towards neuronal cell survival.
authors.affiliation: Klinik für Anesthesiologie, Technische Universität München, Munich, Germany. eberspaecher@ucdavis.edu
authors.forename: E
authors.initials: E
authors.lastname: Eberspächer
authors.forename: C
authors.initials: C
authors.lastname: Werner
authors.forename: K
authors.initials: K
authors.lastname: Engelhard
authors.forename: M
authors.initials: M
authors.lastname: Pape
authors.forename: L
authors.initials: L
authors.lastname: Laacke
authors.forename: D
authors.initials: D
authors.lastname: Winner
authors.forename: R
authors.initials: R
authors.lastname: Hollweck
authors.forename: P
authors.initials: P
authors.lastname: Hutzler
authors.forename: E
authors.initials: E
authors.lastname: Kochs
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D015415:Biomarkers; D015153:Blotting, Western; D002545:Brain Ischemia / Q000378:metabolism / Q000473:pathology*; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D016923:Cell Death; D004789:Enzyme Activation; D005455:Fluorescent Antibody Technique; D006439:Hemodynamics; D006624:Hippocampus / Q000473:pathology; D007036:Hypothermia, Induced; D008297:Male; D009336:Necrosis; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009474:Neurons / Q000473:pathology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015427:Reperfusion Injury / Q000378:metabolism / Q000473:pathology*
chemical_list: D015415:Biomarkers; D009419:Nerve Tissue Proteins; C505410:Casp3 protein, rat; D053148:Caspase 3; D020169:Caspases