title: Inhibitory effect of p53 upregulated modulator of apoptosis targeting siRNA on hypoxia/reoxygenation-induced cardiomyocyte apoptosis in rats.
abstract: OBJECTIVE
  The effect of p53 upregulated modulator of apoptosis (PUMA) in hypoxia/reoxygenation-induced cardiomyocyte injuries in rats was investigated.
  
  
  METHODS
  PUMA-targeting (si-PUMA) and scramble siRNAs were designed and transfected into primarily rat cardiomyocytes in vitro.
  
  
  RESULTS
  RT-PCR and Western blot analysis showed that 50 nmol/l of si-PUMA can specifically inhibit PUMA expression. MTT assay and lactate dehydrogenase activity detection showed that the cell survival rate in the si-PUMA group was enhanced and that the lactate dehydrogenase enzymatic activity dramatically decreased compared with the control group (p < 0.01). Spectrophotometry, as well as annexin V and propidium iodide staining, combined with flow cytometry, revealed that caspase-3 activity in the si-PUMA group was downregulated and the apoptotic rate was decreased (p < 0.01). RT-PCR also showed that Bax expression was downregulated and Bcl-2 expression was upregulated in the si-PUMA group, compared with the control group (p < 0.05). si-PUMA protects cardiomyocytes from apoptosis.
  
  
  CONCLUSIONS
  PUMA mediates hypoxia/reoxygenation-induced cardiomyocyte apoptosis, which can be a potential target of gene therapy for ischemia/reperfusion cardiomyocyte injuries.
authors: Department of Biochemistry and Molecular Biology, Basic Medical College of Nanchang University, Nanchang, China.
authors: Shuo
authors: S
authors: Tu
authors: Zhuo-Qi
authors: ZQ
authors: Liu
authors: Jing-Jing
authors: JJ
authors: Fu
authors: Wei-Feng
authors: WF
authors: Zhu
authors: Da-Ya
authors: DY
authors: Luo
authors: Fu-Sheng
authors: FS
authors: Wan
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism / Q000502:physiology*; D053148:Caspase 3 / Q000378:metabolism; D049108:Cell Enlargement; D015687:Cell Hypoxia / Q000502:physiology; D002470:Cell Survival; D002478:Cells, Cultured; D015536:Down-Regulation; D032383:Myocytes, Cardiac / Q000201:enzymology / Q000502:physiology*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D034741:RNA, Small Interfering; D051381:Rats; D015427:Reperfusion Injury / Q000503:physiopathology; D014162:Transfection; D051028:bcl-2-Associated X Protein / Q000378:metabolism
chemical_list: D051017:Apoptosis Regulatory Proteins; C494828:Bax protein, rat; C533952:Bbc3 protein, rat; D019253:Proto-Oncogene Proteins c-bcl-2; D034741:RNA, Small Interfering; D051028:bcl-2-Associated X Protein; D053148:Caspase 3