Mutated IkappaBalpha represses proliferation of immortalized neural progenitor cells and prevents their apoptosis after oxygen-glucose deprivation.
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.
Zhu
Chang
C
Department of Anesthesiology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China. zhchag@yahoo.com.cn
Liu
Zhiheng
Z
Gui
Lingli
L
Yao
Wenlong
W
Qian
Wei
W
Zhang
Chuanhan
C
D000818:Animals; D000831:Animals, Newborn; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D015153:Blotting, Western; D049109:Cell Proliferation; D002478:Cells, Cultured; D005947:Glucose / Q000494:pharmacology; D051550:I-kappa B Kinase / Q000235:genetics* / Q000378:metabolism; D007150:Immunohistochemistry; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D009154:Mutation; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D010100:Oxygen / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D014162:Transfection
D016328:NF-kappa B; D012333:RNA, Messenger; D007770:L-Lactate Dehydrogenase; D051550:I-kappa B Kinase; D005947:Glucose; D010100:Oxygen