title: PPARγ-mediated advanced glycation end products regulate neural stem cell proliferation but not neural differentiation through the BDNF-CREB pathway.
abstract: OBJECTIVE
  To investigate the roles of PPARγ in advanced glycation end product (AGE)-mediated characteristics of neural stem cells (NSCs) and the molecular mechanisms of action.
  
  
  METHODS
  We prepared pLentiLox3.7 lentiviral vectors expressing short hairpin RNA (shRNA) against PPARγ and transduced NSCs. MTT absorbance and cell counts were used to assay cell growth, and cell differentiation was analysed by confocal laser-scanning and western blots for the expression of MAP2/nestin. The protein and gene expression of the BDNF-CREB pathway components were examined by western blotting and real-time PCR.
  
  
  RESULTS
  Immunoblot analysis indicated that shRNA delivered by lentiviral vectors silenced PPARγ expression in NSCs. The proliferation of NSCs and expression of BDNF pathway components dropped in AGE-BSA culture medium (400 mg/L and 200 mg/L) on Day 3 and Day 7, respectively (all P<0.001). PPARγ-silenced NSCs exhibited a significant increase in cell growth and expression of BDNF pathway components compared with NSCs incubated with AGE-BSA (all P<0.001). Immunocytochemistry and western blotting analysis showed that AGE-BSA (400 mg/L) induced a significant decrease in the expression of MAP2 both in NSCs and PPARγ-silenced NSCs, as standardised by nestin. There was no significant difference between NSCs and PPARγ-silenced NSCs in the presence of AGE-BSA.
  
  
  CONCLUSIONS
  PPARγ plays roles in the AGE-mediated regulation of NSC proliferation but not neural differentiation through the BDNF-CREB pathway.
authors.affiliation: The Department of Endocrinology, Affiliated ZhongDa Hospital of Southeast University, No. 87 DingJiaQiao Road, Nanjing 210009, PR China. gyjwsh@yahoo.cn
authors.forename: Shao-hua
authors.initials: SH
authors.lastname: Wang
authors.forename: Yi-jing
authors.initials: YJ
authors.lastname: Guo
authors.forename: Yang
authors.initials: Y
authors.lastname: Yuan
authors.forename: Ling
authors.initials: L
authors.lastname: Li
authors.forename: Feng-fei
authors.initials: FF
authors.lastname: Li
authors.forename: Kuan-ping
authors.initials: KP
authors.lastname: Ye
authors.forename: Yan
authors.initials: Y
authors.lastname: Huang
mesh_terms: D000818:Animals; D019208:Brain-Derived Neurotrophic Factor / Q000502:physiology*; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D003470:Culture Media; D017362:Cyclic AMP Response Element-Binding Protein / Q000502:physiology*; D017127:Glycation End Products, Advanced / Q000502:physiology*; D058953:Neural Stem Cells / Q000166:cytology*; D047495:PPAR gamma / Q000502:physiology*; D034741:RNA, Small Interfering / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D019208:Brain-Derived Neurotrophic Factor; D003470:Culture Media; D017362:Cyclic AMP Response Element-Binding Protein; D017127:Glycation End Products, Advanced; D047495:PPAR gamma; D034741:RNA, Small Interfering