title: Advanced glycation end product-induced astrocytic differentiation of cultured neurospheres through inhibition of Notch-Hes1 pathway-mediated neurogenesis.
abstract: This study aims to investigate the roles of the Notch-Hes1 pathway in the advanced glycation end product (AGE)-mediated differentiation of neural stem cells (NSCs). We prepared pLentiLox3.7 lentiviral vectors that express short hairpin RNA (shRNA) against Notch1 and transfected it into NSCs. Cell differentiation was analyzed under confocal laser-scanning microscopy. The percentage of neurons and astrocytes was quantified by normalizing the total number of TUJ1+ (Neuron-specific class III β-tubulin) and GFAP+ (Glial fibrillary acidic protein) cells to the total number of Hoechst 33342-labeled cell nuclei. The protein and gene expression of Notch-Hes1 pathway components was examined via western blot analysis and real-time PCR. After 1 week of incubation, we found that AGE-bovine serum albumin (BSA) (400 μg/mL) induced the astrocytic differentiation of cultured neurospheres and inhibited neuronal formation. The expression of Notch-Hes1 pathway components was upregulated in the cells in the AGE-BSA culture medium. Immunoblot analysis indicated that shRNA silencing of Notch1 expression in NSCs significantly increases neurogenesis and suppresses astrocytic differentiation in NSCs incubated with AGE-BSA. AGEs promote the astrocytic differentiation of cultured neurospheres by inhibiting neurogenesis through the Notch-Hes1 pathway, providing a potential therapeutic target for hyperglycemia-related cognitive deficits.
authors.forename: Yijing
authors.initials: Y
authors.lastname: Guo
authors.forename: Pin
authors.initials: P
authors.lastname: Wang
authors.forename: Haixia
authors.initials: H
authors.lastname: Sun
authors.forename: Rongrong
authors.initials: R
authors.lastname: Cai
authors.forename: Wenqing
authors.initials: W
authors.lastname: Xia
authors.affiliation: Department of Neurology, Affiliated ZhongDa Hospital of Southeast University, No.87 DingJiaQiao Road, Nanjing 210009, China. gyjwsh@gmail.com.
authors.forename: Shaohua
authors.initials: S
authors.lastname: Wang
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000166:cytology* / Q000378:metabolism; D051792:Basic Helix-Loop-Helix Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002417:Cattle; D002478:Cells, Cultured; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D017127:Glycation End Products, Advanced / Q000494:pharmacology*; D018398:Homeodomain Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D058953:Neural Stem Cells / Q000166:cytology / Q000187:drug effects*; D055495:Neurogenesis / Q000187:drug effects*; D016716:PC12 Cells; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D051381:Rats; D051880:Receptors, Notch / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000494:pharmacology*; D000072056:Transcription Factor HES-1; D014404:Tubulin / Q000378:metabolism; D015854:Up-Regulation / Q000187:drug effects
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D005904:Glial Fibrillary Acidic Protein; D017127:Glycation End Products, Advanced; D018398:Homeodomain Proteins; D034741:RNA, Small Interfering; D051880:Receptors, Notch; D012710:Serum Albumin, Bovine; D000072056:Transcription Factor HES-1; D014404:Tubulin; C489735:Hes1 protein, rat; C488416:Tubb3 protein, rat; C081628:advanced glycation end products-bovine serum albumin