title: Deep hypothermia attenuates microglial proliferation independent of neuronal death after prolonged cardiac arrest in rats.
abstract: BACKGROUND
  Conventional resuscitation of exsanguination cardiac arrest (CA) victims is generally unsuccessful. Emergency preservation and resuscitation is a novel approach that uses an aortic flush to induce deep hypothermia during CA, followed by delayed resuscitation with cardiopulmonary bypass. Minocycline has been shown to be neuroprotective across a number of brain injury models via attenuating microglial activation. We hypothesized that deep hypothermia and minocycline would attenuate neuronal death and microglial activation and improve outcome after exsanguination CA in rats.
  
  
  METHODS
  Using isoflurane anesthesia, rats were subjected to a lethal hemorrhagic shock. After 5 min of no flow, hypothermia was induced with an aortic flush. Three groups were studied: ice-cold (IC) flush, room-temperature (RT) flush, and RT flush followed by minocycline treatment (RT-M). After 20 min of CA, resuscitation was achieved via cardiopulmonary bypass. Survival, Overall Performance Category (1 = normal, 5 = death), Neurologic Deficit Score (0%-10% = normal, 100% = max deficit), neuronal death (Fluoro-Jade C), and microglial proliferation (Iba1 immunostaining) in hippocampus were assessed at 72 h.
  
  
  RESULTS
  Rats in the IC group had lower tympanic temperature during CA versus other groups (IC, 20.9 degrees C +/- 1.3 degrees C; RT, 28.4 degrees C +/- 0.6 degrees C; RT-M, 28.3 degrees C +/- 0.7 degrees C; P < 0.001). Although survival was similar in all groups (RT, 6/9; IC, 6/7; RT-M, 6/11), neurological outcome was better in the IC group versus other groups (Overall Performance Category: IC, 1 +/- 1; RT, 3 +/- 1; RT-M, 2 +/- 1; P < 0.05; Neurologic Deficit Score: IC, 8% +/- 9%; RT, 55% +/- 19%; RT-M, 27% +/- 16%; P < 0.05). Histological damage assessed in survivors showed selective neuronal death in CA1 and dentate gyrus, similar in all groups (P = 0.15). In contrast, microglial proliferation was attenuated in the IC group versus all other groups (P < 0.01).
  
  
  CONCLUSIONS
  Deeper levels of hypothermia induced by the IC versus RT flush resulted in better neurological outcome in survivors. Surprisingly, deep hypothermia attenuated microglial activation but not hippocampal neuronal death. Minocycline had modest benefit on neurologic outcome in survivors but did not attenuate microglial activation in brain. Our findings suggest a novel effect of deep hypothermia on microglial proliferation during exsanguination CA.
authors.affiliation: Safar Center for Resuscitation Research, University of Pittsburgh, Pittsburgh, PA 15260, USA. drabekt@anes.upmc.edu
authors.forename: Tomas
authors.initials: T
authors.lastname: Drabek
authors.forename: Samuel A
authors.initials: SA
authors.lastname: Tisherman
authors.forename: Lauren
authors.initials: L
authors.lastname: Beuke
authors.forename: Jason
authors.initials: J
authors.lastname: Stezoski
authors.forename: Keri
authors.initials: K
authors.lastname: Janesko-Feldman
authors.forename: Manuella
authors.initials: M
authors.lastname: Lahoud-Rahme
authors.forename: Patrick M
authors.initials: PM
authors.lastname: Kochanek
mesh_terms: D000818:Animals; D001011:Aorta / Q000473:pathology; D001835:Body Weight; D016887:Cardiopulmonary Resuscitation / Q000379:methods; D049109:Cell Proliferation; D006323:Heart Arrest / Q000503:physiopathology*; D006339:Heart Rate; D007036:Hypothermia, Induced / Q000379:methods; D008297:Male; D017628:Microglia / Q000187:drug effects / Q000473:pathology*; D008911:Minocycline / Q000494:pharmacology; D009474:Neurons / Q000187:drug effects / Q000473:pathology*; D018696:Neuroprotective Agents / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D018696:Neuroprotective Agents; D008911:Minocycline