title: [Adenovirus vector mediated human 14-3-3 γ gene transfer protects dopaminergic cells against rotenone-induced injury].
abstract: OBJECTIVE
  To explore the protective effect of human 14-3-3 γ gene transfer on dopaminergic cells against rotenone-induced injury.
  
  
  METHODS
  Adenovirus vector carrying the gene of 14-3-3 γ (Ad/14-3-3 γ) was employed to transfect PC12 cells. Then the cells were exposed to rotenone as a model of Parkinson's disease. Methyl thiazolyl tetrazolium (MTT) was used to assay the viability of PC12 cells. The 4',6-diamidino-2-phenylindole (DAPI) staining was used to analyze the apoptotic ratio of PC12 cells among the groups of control, Ad/14-3-3 γ, Ad-null and Rotenone. And high performance liquid chromatography (HPLC) was performed to detect the secreting functions of PC12 cells. The aggregates of α-Synuclein protein were detected under confocal microscopy.
  
  
  RESULTS
  MTT showed that the cell absorbance A(570) of Ad/14-3-3 γ group (0.46 ± 0.09) was higher than that of Ad-null group (0.19 ± 0.08) and rotenone group (0.16 ± 0.07), but lower than that of normal control (0.63 ± 0.11), (all P < 0.01); HPLC-ECD showed that the levels of dopamine (189 ± 11) ng/ml and noradrenalin (55 ± 8) ng/ml in the culture fluid of Ad/14-3-3 γ group were higher than those of Ad-null group (79 ± 12, 38 ± 7) ng/ml and rotenone group (81 ± 13, 39 ± 7) ng/ml (all P < 0.01). DAPI staining showed that cell apoptosis ratio of group Ad/14-3-3 γ (27% ± 64%) was lower than that of group Ad-null (53% ± 10%) and rotenone group (56% ± 12%, P < 0.01), but higher than that of control group (10% ± 5%, P < 0.01). Under confocal microscopy, the aggregates of α-synuclein protein in PC12 cells were detected more in Ad-null group and rotenone group than that in Ad/14-3-3 γ group.
  
  
  CONCLUSIONS
  Gene transfer of Ad/14-3-3 γ has a protective effect on dopaminergic cells against rotenone-induced injury.
authors.affiliation: Department of Neurology, Hainan Medical College, Haikou, China.
authors.forename: Xiao-wu
authors.initials: XW
authors.lastname: Chen
authors.forename: Tan
authors.initials: T
authors.lastname: Wang
authors.forename: Kun-xiong
authors.initials: KX
authors.lastname: Yuan
authors.forename: Yuan-jiang
authors.initials: YJ
authors.lastname: Wang
authors.forename: Shu-rong
authors.initials: SR
authors.lastname: Wang
authors.forename: Sheng-gang
authors.initials: SG
authors.lastname: Sun
authors.forename: Zhi-bin
authors.initials: ZB
authors.lastname: Chen
mesh_terms: D048948:14-3-3 Proteins / Q000235:genetics*; D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D002470:Cell Survival; D059290:Dopaminergic Neurons / Q000187:drug effects* / Q000378:metabolism*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D016716:PC12 Cells; D051381:Rats; D012402:Rotenone / Q000009:adverse effects*
chemical_list: D048948:14-3-3 Proteins; D012402:Rotenone