title: Effect of apoptosis in neural stem cells treated with sevoflurane.
abstract: BACKGROUND
  At present, sevoflurane inhalation anesthesia used on infants is well-known. But long-time exposure to inhalation anesthetic could cause neurologic disorder, especially nerve degeneration in infant and developing brain. The central nervous system degeneration of infants could affect the memory and cognitive function. γ-Aminobutyric acid (GABA) is a known inhibitory neurotransmitter in central nervous system. Inhalation anesthetic sevoflurane may activate GABAA receptor to inhibit central nervous system, leading to apoptosis of neural degeneration, cognitive dysfunction in the critical period of brain development.
  
  
  METHODS
  Neural stem cells were derived from Wistar embryos, cultured in vitro. Third generation of neural stem cells were randomly divided into four groups according to cultured suspension: Sevoflurane group (Group S), GABAA receptor antagonists, Bicuculline group (Group B), Sevoflurane + GABAA receptor antagonists, Bicuculline group (Group S + B), dimethyl sulphoxide (DMSO) group (Group D). Group B and Group D did not receive sevoflurane preconditioning. Group S and Group S + B were pretreated with 1 minimum alveolar concentration (MAC) sevoflurane for 0 h, 3 h, 6 h, and 12 h. Group S + B and Group B were pretreated with bicuculline (10 uM). Group D was treated with DMSO (10 uL/mL). After treatments above, all groups were cultured for 48 h. Then we measured the cells viability by Cell Counting Kit (CCK-8) assay, cytotoxicity by Lactate Dehydrogenase (LDH) assay, apoptosis ratio with Annexin V/propidium iodide (PI) staining by flow cytometry, and the expression of GABAAR, anti-apoptotic protein Bcl-2, pro-apoptotic protein Bax and Caspase-3 by western blotting.
  
  
  RESULTS
  After exposing to sevoflurane for 0 h, 3 h, 6 h, and 12 h with 1MAC, we found that cell viability obviously decreased and cytotoxicity increased in time-dependent way. And Annexin V/PI staining indicated increased apoptosis ratio by flow cytometry. The protein level of GABAA receptor, pro-apoptotic protein Bax and apoptosis protein Caspase-3 increased; while anti-apoptotic protein Bcl-2 decreased. And bicuculline could reverse all detrimental results caused by sevoflurane.
  
  
  CONCLUSIONS
  Sevoflurane can inhibit the central nervous system by activating GABAA, resulting in apoptosis of neural stem cells, thus leading to the NSCs degeneration.
authors.affiliation: Department of Anesthesiology, Dezhou People's Hospital, Dezhou, Shandong China ; School of Medicine, Shandong University, Ji'nan, Shandong China.
authors.forename: Jianlei
authors.initials: J
authors.lastname: Qiu
authors.affiliation: Department of Anesthesiology, Shandong Provincial Qianfoshan Hospital, Ji'nan, Shandong China.
authors.forename: Pengcai
authors.initials: P
authors.lastname: Shi
authors.affiliation: Department of Anesthesiology, Jiaozhou Central Hospital of Qingdao, Qingdao, Shandong China.
authors.forename: Wude
authors.initials: W
authors.lastname: Mao
authors.affiliation: Department of Anesthesiology, Dezhou People's Hospital, Dezhou, Shandong China.
authors.forename: Yuyi
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Department of Emergency, Dezhou People's Hospital, Dezhou, Shandong China.
authors.forename: Wenshuai
authors.initials: W
authors.lastname: Liu
authors.affiliation: School of Medicine, Shandong University, Ji'nan, Shandong China ; Department of Anesthesiology, Shandong Provincial Qianfoshan Hospital, Ji'nan, Shandong China.
authors.forename: Yuelan
authors.initials: Y
authors.lastname: Wang
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001640:Bicuculline / Q000494:pharmacology; D053148:Caspase 3 / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D008738:Methyl Ethers / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D058953:Neural Stem Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis; D051381:Rats; D011963:Receptors, GABA-A / Q000378:metabolism; D000077149:Sevoflurane; D051028:bcl-2-Associated X Protein / Q000378:metabolism
chemical_list: D008738:Methyl Ethers; D019253:Proto-Oncogene Proteins c-bcl-2; D011963:Receptors, GABA-A; D051028:bcl-2-Associated X Protein; D000077149:Sevoflurane; D053148:Caspase 3; D001640:Bicuculline
keywords: Apoptosis; Neural Stem Cells; Sevoflurane; γ-Aminobutyric acid