Mutated IkappaBalpha represses proliferation of immortalized neural progenitor cells and prevents their apoptosis after oxygen-glucose deprivation

Brain Res. 2008 Dec 9:1244:24-31. doi: 10.1016/j.brainres.2008.08.003. Epub 2008 Aug 12.

Abstract

Endogenic and transplanted neural progenitor cells (NPCs) can be activated by cerebral ischemia and take part in the regeneration of neural function. NF-kappaB was found activated in the same pathology procedure and was assumed to play a crucial role in regulating NPCs' physiology. But it is still not clear whether NF-kappaB is activated in NPCs in cerebral ischemia and what is the effect of NF-kappaB on NPCs when activated. Our previous work generated immortalized neural progenitor cells (INPCs) to provide simulation for NPCs. Then pcDNA3.1 transfected INPCs (INPCs/pcDNA3.1) and mutated IkappaBalpha gene transfected INPCs (INPCs/IkappaBalphaM) were generated. By western blotting and electrophoretic mobility shift assay mutated IkappaBalpha was found expressed in INPCs/IkappaBalphaM and repressed the activity of NF-kappaB in INPCs. No difference in the differentiation of INPCs/pcDNA3.1 and INPCs/IkappaBalphaM was found by western blotting and immunocytochemistry. Detected by MTT assay INPCs/IkappaBalphaM had a lower proliferation rate under normal conditions. The apoptosis rate and lactate dehydrogenase activity in the medium of INPCs/IkappaBalphaM were lower than INPCs/pcDNA3.1 after oxygen-glucose deprivation. Some NF-kappaB-driven cytokines were observed down-regulated in INPCs/IkappaBalphaM by real-time reverse transcription polymerase chain reaction. In our research NF-kappaB was found activated in INPCs after oxygen-glucose deprivation. NF-kappaB activity down-regulation represses proliferation of INPCs and improves their tolerance to oxygen-glucose deprivation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Newborn
  • Apoptosis / drug effects
  • Apoptosis / physiology*
  • Blotting, Western
  • Cell Proliferation*
  • Cells, Cultured
  • Glucose / pharmacology
  • I-kappa B Kinase / genetics*
  • I-kappa B Kinase / metabolism
  • Immunohistochemistry
  • L-Lactate Dehydrogenase / metabolism
  • Mutation
  • NF-kappa B / genetics
  • NF-kappa B / metabolism
  • Neurons / cytology
  • Neurons / metabolism
  • Neurons / physiology*
  • Oxygen / pharmacology
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Rats
  • Reverse Transcriptase Polymerase Chain Reaction
  • Stem Cells / cytology
  • Stem Cells / metabolism
  • Stem Cells / physiology*
  • Transfection

Substances

  • NF-kappa B
  • RNA, Messenger
  • L-Lactate Dehydrogenase
  • I-kappa B Kinase
  • Glucose
  • Oxygen