Mild postischemic hypothermia prolongs the time window for gene therapy by inhibiting cytochrome C release

Stroke. 2004 Feb;35(2):572-7. doi: 10.1161/01.STR.0000110787.42083.58. Epub 2004 Jan 15.

Abstract

Background and purpose: We showed previously that Bcl-2 overexpression with the use of herpes simplex viral (HSV) vectors improved striatal neuron survival when delivered 1.5 hours after stroke but not when delivered 5 hours after stroke onset. Here we determine whether hypothermia prolongs the therapeutic window for gene therapy.

Methods: Rats were subjected to focal ischemia for 1 hour. Hypothermia (33 degrees C) was induced 2 hours after insult and maintained for 3 hours. Five hours after ischemia onset, HSV vectors expressing Bcl-2 plus beta-gal or beta-gal alone were injected into each striatum. Rats were killed 2 days later.

Results: Striatal neuron survival of Bcl-2-treated, hypothermic animals was improved 2- to 3-fold over control-treated, hypothermic animals and Bcl-2-treated, normothermic animals. Neuron survival among normothermic, Bcl-2-treated animals was not different from control normothermics or control hypothermics. Double immunostaining of cytochrome c and beta-gal demonstrated that Bcl-2 plus hypothermia significantly reduced cytochrome c release.

Conclusions: Postischemic mild hypothermia extended the time window for gene therapy neuroprotection using Bcl-2 and reduced cytochrome c release.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Brain Ischemia / complications
  • Brain Ischemia / pathology
  • Brain Ischemia / physiopathology
  • Brain Ischemia / therapy*
  • Cell Survival / drug effects
  • Corpus Striatum / pathology
  • Cytochromes c / antagonists & inhibitors*
  • Cytochromes c / metabolism
  • Disease Models, Animal
  • Genes, Reporter
  • Genetic Therapy / methods*
  • Genetic Vectors / administration & dosage
  • Genetic Vectors / genetics
  • Hypothermia, Induced / methods*
  • Neurons / drug effects
  • Neurons / metabolism
  • Neurons / pathology
  • Proto-Oncogene Proteins c-bcl-2 / biosynthesis
  • Proto-Oncogene Proteins c-bcl-2 / genetics
  • Proto-Oncogene Proteins c-bcl-2 / pharmacology
  • Rats
  • Rats, Sprague-Dawley
  • Simplexvirus / genetics
  • Stroke / complications
  • Stroke / pathology
  • Stroke / physiopathology
  • Stroke / therapy*
  • Time Factors
  • beta-Galactosidase / biosynthesis
  • beta-Galactosidase / genetics

Substances

  • Proto-Oncogene Proteins c-bcl-2
  • Cytochromes c
  • beta-Galactosidase