title: [Effect of antisense oligonucleotide against Smac/DIABLO on inhibition of hydrogen peroxide induced myocardial apoptosis of neonatal rats].
abstract: OBJECTIVE
  To investigate the effects of antisense phosphorothioate oligonucleotides against Smac/DIABLO (asODN) on hydrogen peroxide (H2O2) induced myocardial apoptosis in neonatal rats.
  
  
  METHODS
  Primary myocardial cells from neonatal rats were cultured in vitro, and randomly divided into A (normal control, without transfection), B (with treatment of single liposome), C (with transfection of scrODN), D (with transfection of asODN), E (with H2O2, stimulation), F (with H2O2 stimulation after scrODN transfection), and G (with H2O2 stimulation after asODN transfection) groups. The expression of asODN mRNA and protein were determined with RT-PCR and Western blotting, respectively. The changes in cellular nuclear morphology were observed with 33258 fluorescent staining, and the percentage of nuclear apoptosis was calculated. DNA fragmentation was observed by agarose gel electrophoresis. Activation of caspase-3 and caspase-9 were evaluated by caspase colorimetric analysis kit.
  
  
  RESULTS
  The expression of Smac/DIABLO mRNA and protein was obviously inhibited by asODN, which was about 80% percent lower than the protein level in A,B and C groups, but there was no difference noted among A,B and C groups( P > 0.05). Not only the nuclear apoptotic percentage, but also the activity of caspase-3 and caspase-9 in A, C and D groups were in very low levels. But these indices in G group 24 hours after H2O2 stimulation were obviously lower than that in E and F groups [the nuclear apoptotic percentage were (19 +/- 5) %, (52 +/- 3) %, (55 +/- 5) %, respectively, P < 0.01)]. The DNA ladders in G group were also decreased markedly.
  
  
  CONCLUSIONS
  Myocardial apoptosis induced by H2O2 can be inhibited by asODN in rat.
authors.affiliation: Departnent of Burns and Plastic Surgery, Xiangya Hospital, Central Southern University, Changsha 410008, PR China.
authors.forename: Peng-fei
authors.initials: PF
authors.lastname: Liang
authors.forename: Xiao-yuan
authors.initials: XY
authors.lastname: Huang
authors.forename: Jian-hong
authors.initials: JH
authors.lastname: Long
authors.forename: Mu-zhang
authors.initials: MZ
authors.lastname: Xiao
authors.forename: Xing-hua
authors.initials: XH
authors.lastname: Yang
authors.forename: Pi-hong
authors.initials: PH
authors.lastname: Zhang
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D051017:Apoptosis Regulatory Proteins; D002352:Carrier Proteins / Q000378:metabolism*; D053148:Caspase 3 / Q000378:metabolism; D053453:Caspase 9 / Q000378:metabolism; D002478:Cells, Cultured; D006861:Hydrogen Peroxide; D024101:Mitochondrial Proteins / Q000378:metabolism*; D032383:Myocytes, Cardiac / Q000187:drug effects* / Q000378:metabolism; D020319:Oligodeoxyribonucleotides, Antisense / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar
chemical_list: D051017:Apoptosis Regulatory Proteins; D002352:Carrier Proteins; C489013:DIABLO protein, rat; D024101:Mitochondrial Proteins; D020319:Oligodeoxyribonucleotides, Antisense; D006861:Hydrogen Peroxide; D053148:Caspase 3; D053453:Caspase 9