title: Nerve Growth Factor Protects Against Alcohol-Induced Neurotoxicity in PC12 Cells via PI3K/Akt/mTOR Pathway.
abstract: OBJECTIVE
  To study whether autophagy participates in the neuroprotective effect of nerve growth factor (NGF) on neurons treated with alcohol.
  
  
  METHODS
  The autophagy-related markers were used to explore the role of autophagy in PC12 cells exposed to alcohol or pre-incubated with NGF before initiating the treatment with alcohol (100 mM; 6 h). PC12 cells were pre-incubated with 3-methyladenine (3-MA) (10 mM; 1 h) or rapamycin (100 nM; 1 h) before co-incubated with alcohol (100 mM; 6 h) in order to investigate the relationship between apoptosis and autophagy. PC12 cells were pre-incubated with LY294002 (50 μM; 30 min) before co-incubated with NGF and alcohol in order to analyze the protein expression of PI3K/Akt/mTOR pathway via western blotting.
  
  
  RESULTS
  By methylthiazoltetrazolium, western blotting and flow cytometry assays, we found that cell viability decreased in a dose- and time-dependent manner after treatment with alcohol in PC12 cells. As cells were exposed to alcohol, the levels of LC3-II proteins became elevated, likewise, pre-treatment with 3-methyladenine (3-MA, an autophagic inhibitor) or rapamycin (an autophagic inducer) resulted in an increased or decreased percentage of apoptosis in contrast to other alcohol-treated groups, respectively. NGF markedly increased LC3-II production after treatment with alcohol in a dose-dependent manner. Moreover, NGF remarkably attenuated the phosphorylation effect of alcohol exposure on PI3K/Akt/mTOR pathway, which was suppressed by LY294002 (Akt inhibitor).
  
  
  CONCLUSIONS
  NGF protects against alcohol-induced neurotoxicity via PI3K/Akt/mTOR pathway.
  
  
  OBJECTIVE
  In this study, we chose the PC12 cell line as a neuronal model, and our results demonstrate that nerve growth factor can induce autophagy with the neuroprotective effect and regulatory mechanisms of alcohol-induced autophagy in PC12 cells.
authors.affiliation: The First Affiliated Hospital of Harbin Medical University, 23 Youzheng Street, Nangang District, Harbin, Heilongjiang 150001, PR China.
authors.forename: Liping
authors.initials: L
authors.lastname: Liu
authors.affiliation: School of Life Science and Technology, Harbin Institute of Technology, 2 Yikuang Street, Nangang District, Harbin, Heilongjiang 150080, PR China.
authors.forename: Tiedong
authors.initials: T
authors.lastname: Sun
authors.affiliation: The First Affiliated Hospital of Harbin Medical University, 23 Youzheng Street, Nangang District, Harbin, Heilongjiang 150001, PR China.
authors.forename: Feng
authors.initials: F
authors.lastname: Xin
authors.affiliation: School of Life Science and Technology, Harbin Institute of Technology, 2 Yikuang Street, Nangang District, Harbin, Heilongjiang 150080, PR China.
authors.forename: Wei
authors.initials: W
authors.lastname: Cui
authors.affiliation: School of Life Science and Technology, Harbin Institute of Technology, 2 Yikuang Street, Nangang District, Harbin, Heilongjiang 150080, PR China.
authors.forename: Jianquan
authors.initials: J
authors.lastname: Guo
authors.affiliation: The First Affiliated Hospital of Harbin Medical University, 23 Youzheng Street, Nangang District, Harbin, Heilongjiang 150001, PR China hujian0451@163.com.
authors.forename: Jian
authors.initials: J
authors.lastname: Hu
mesh_terms: D000818:Animals; D001343:Autophagy / Q000187:drug effects / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D000431:Ethanol / Q000633:toxicity*; D051379:Mice; D020932:Nerve Growth Factor / Q000494:pharmacology*; D018696:Neuroprotective Agents / Q000494:pharmacology*; D016716:PC12 Cells; D058539:Phosphatidylinositol 3-Kinase / Q000378:metabolism*; D000081082:Phosphoinositide-3 Kinase Inhibitors; D051057:Proto-Oncogene Proteins c-akt / Q000037:antagonists & inhibitors / Q000378:metabolism*; D051381:Rats; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D058570:TOR Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*
chemical_list: D018696:Neuroprotective Agents; D000081082:Phosphoinositide-3 Kinase Inhibitors; D000431:Ethanol; D020932:Nerve Growth Factor; D058539:Phosphatidylinositol 3-Kinase; D051057:Proto-Oncogene Proteins c-akt; D058570:TOR Serine-Threonine Kinases