title: [Effects of neuroactive steroid allopregnanolone on the damage of cortical neurons].
abstract: OBJECTIVE
  To investigate the protective mechanism of neuroactive steroid allopregnanolone on N-methyl-D-aspartate (NMDA) induced toxicity in primary mouse cortical neurons.
  
  
  METHODS
  Primary cultured mouse cortical neurons were subjected to allopregnanolone, the expression of beta-aminobutyric acid receptor beta2 subunit (beta2-GABA-R) mRNAs was detected by RT-PCR and Akt phosphorylation was assayed by Western blot using Akt-phosphoserine 473-specific antibody. After the cultured mouse cortical neurons were pretreated with or without allopregnanolone prior to treatment with NMDA , DNA isolated was analyzed by agarose gel electrophoresis and proteins collected were analyzed by Western blot with anti-cleaved-PARP, anti-cleaved caspase-3, and anti-cleaved caspase-9 antibodies.
  
  
  RESULTS
  When cultured mouse cortical neurons were exposed to allopregnanolone both the expression of beta2-GABA-R mRNAs and Akt phosphorylation increased. Allopregnanolone inhibited the NMDA-induced apoptosis and decreased the level of active-PARP, active-caspase-3 and active-caspase-9 notably at a final concentration of 5 x 10(6) mol/L.
  
  
  CONCLUSIONS
  Pretreatment with allopregnanolone may be neuroprotective on NMDA-induced neuronal cells apoptosis by increasing beta2-GABA-R expression and Akt phosphorylation.
authors.affiliation: Department of Molecular Biology, Medical College of Chinese People's Armed Police Force, Tianjing 300162, China. lixianhuiwj@163.com
authors.forename: Xian-Hui
authors.initials: XH
authors.lastname: Li
authors.forename: Xin-Chang
authors.initials: XC
authors.lastname: Zhang
authors.forename: Gang
authors.initials: G
authors.lastname: Wang
authors.forename: Hai-Ling
authors.initials: HL
authors.lastname: Liu
authors.forename: Shi-Hai
authors.initials: SH
authors.lastname: Xia
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D017209:Apoptosis / Q000187:drug effects; D053148:Caspase 3 / Q000378:metabolism; D053453:Caspase 9 / Q000378:metabolism; D002540:Cerebral Cortex / Q000166:cytology*; D051379:Mice; D016202:N-Methylaspartate / Q000037:antagonists & inhibitors / Q000633:toxicity*; D009474:Neurons / Q000166:cytology*; D018696:Neuroprotective Agents / Q000494:pharmacology*; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D011280:Pregnanolone / Q000494:pharmacology*; D061251:Primary Cell Culture; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D018080:Receptors, GABA-B / Q000235:genetics / Q000378:metabolism
chemical_list: D018696:Neuroprotective Agents; D012333:RNA, Messenger; D018080:Receptors, GABA-B; D016202:N-Methylaspartate; D011280:Pregnanolone; C499321:Parp1 protein, mouse; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases; C505409:Casp3 protein, mouse; C506434:Casp9 protein, mouse; D053148:Caspase 3; D053453:Caspase 9