title: [Suppressive effects of par-4 antisense oligodeoxynucleotide on up-regulation of intracellular calcium concentration in PC12 cell induced by glutamate and its anti-apoptosis effects].
abstract: OBJECTIVE
  To investigate the uppressive effects of par-4 antisense oligodeoxynucleotide on the up-regulation of intracellular calcium concentration in PC12 cell induced by glutamate and its anti-apoptosis effects.
  
  
  METHODS
  Cationic lipid-mediated par-4 antisense oligodeoxynucleotide (par-4-AS-ODN) was transfected into PC12 cells and followed by glutamate for treatment. Mismatch oligodeoxy-nucleotide (MS-ODN) was used as the control. Morphological assessment and evaluation of the anti-apoptosis effects of par-4-AS-ODN on PC12 cells were performed by laser scanning confocal microscopy by double staining of the cells with Hoechest 33258/propidium iodide (Hoe/PI) and flow cytometry respectively. The mRNA and protein levels of calpain 10 and par-4 were measured by RT-PCR and Western blot. Intracellular calcium concentration was determined by using laser scanning confocal microscope with Fura-2/AM as the fluorescent dye.
  
  
  RESULTS
  Par-4-AS-ODN repress the increase of par-4 protein in PC12 cell (52.3 +/- 5.0 vs 90.0 +/- 3.2, < 0.01). Par-4-AS-ODN significantly inhibited the apoptosis of PC12 cells induced by glutamate (53% vs 31%, < 0.01). Par-4-AS-ODN significantly suppress the up-regulation of intracellular calcium concentration in PC12 cells induced by glutamate (Rate of fluorescent density: 167.9 +/- 32.4 vs 228.8 +/- 36.8, < 0.01). Par-4-AS-ODN inhibited the increase of calpain 10 mRNA in PC12 cells induced by glutamate (46.3 +/- 3.7 vs 34.8 +/- 2.1, < 0.01).
  
  
  CONCLUSIONS
  par-4-AS-ODN enables to inhibit apoptosis of PC12 cells induced by glutamate. The mechanism of the inhibition may be closely related to suppression of the up-regulation of intracellular calcium concentration and calpain transcription expression.
authors.affiliation: Department of Pediatrics, First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China. luchao8009@sohu.com
authors.forename: Chao
authors.initials: C
authors.lastname: Lu
authors.forename: Ji-qing
authors.initials: JQ
authors.lastname: Chen
authors.forename: Sheng-hua
authors.initials: SH
authors.lastname: Wu
authors.forename: Fei
authors.initials: F
authors.lastname: Zhao
authors.forename: Xia
authors.initials: X
authors.lastname: Chi
authors.forename: Xiao-qin
authors.initials: XQ
authors.lastname: Pan
authors.forename: Li
authors.initials: L
authors.lastname: Fei
authors.forename: Mei
authors.initials: M
authors.lastname: Guo
authors.forename: Song-ming
authors.initials: SM
authors.lastname: Huang
authors.forename: Xi-rong
authors.initials: XR
authors.lastname: Guo
authors.forename: Rong-hua
authors.initials: RH
authors.lastname: Chen
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D051017:Apoptosis Regulatory Proteins / Q000096:biosynthesis* / Q000235:genetics; D002118:Calcium / Q000378:metabolism*; D002154:Calpain / Q000096:biosynthesis / Q000235:genetics; D018698:Glutamic Acid / Q000494:pharmacology; D020319:Oligodeoxyribonucleotides, Antisense / Q000494:pharmacology*; D016716:PC12 Cells; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D014162:Transfection; D015854:Up-Regulation
chemical_list: D051017:Apoptosis Regulatory Proteins; D020319:Oligodeoxyribonucleotides, Antisense; D012333:RNA, Messenger; C101296:prostate apoptosis response-4 protein; D018698:Glutamic Acid; D002154:Calpain; C415197:calpain 10; D002118:Calcium