title: Recombinant adeno-associated virus serotype 9 with p65 ribozyme protects H9c2 cells from oxidative stress through inhibiting NF-κB signaling pathway.
abstract: BACKGROUND
  Oxidative stress is a major mechanism underlying the pathogenesis of cardiovascular disease. It can trigger inflammatory cascades which are primarily mediated via nuclear factor-κB (NF-κB). The NF-κB transcription factor family includes several subunits (p50, p52, p65, c-Rel, and Rel B) that respond to myocardial ischemia. It has been proved that persistent myocyte NF-κB p65 activation in heart failure exacerbates cardiac remodeling.
  A recombinant adeno-associated virus serotype 9 carrying enhanced green fluorescent protein and anti-NF-κB p65 ribozyme (AAV9-R65-CMV-eGFP) was constructed. The cells were assessed by MTT assay, Annexin V-propidium iodide dual staining to study apoptosis. The expression of P65 and P50 were assessed by Western blot to investigate the underlying molecular mechanisms.
  
  
  RESULTS
  After stimulation with H2O2 for 6 h, H9c2 cells viability decreased significantly, a large fraction of cells underwent apoptosis. We observed a rescue of H9c2 cells from H2O2-induced apoptosis in pretreatment with AAV9-R65-CMV-eGFP. Moreover, AAV9-R65-CMV-eGFP decreased H2O2-induced P65 expression.
  
  
  CONCLUSIONS
  AAV9-R65-CMV-eGFP protects H9c2 cells from oxidative stress induced apoptosis through down-regulation of P65 expression. These observations indicate that AAV9-R65-CMV-eGFP has the potential to exert cardioprotective effects against oxidative stress, which might be of great importance to clinical efficacy for cardiovascular disease.
authors.affiliation: Xinjiang Key Laboratory of Cardiovascular Disease Research, Urumqi 830054, Xinjiang Uygur Autonomous Region, China ; Laboratory of Functional Experiment, Preclinical Medicine, Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China.
authors.forename: Zhan
authors.initials: Z
authors.lastname: Sun
authors.affiliation: Xinjiang Key Laboratory of Cardiovascular Disease Research, Urumqi 830054, Xinjiang Uygur Autonomous Region, China ; Department of Cardiology, the First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China.
authors.forename: Yi-Tong
authors.initials: YT
authors.lastname: Ma
authors.affiliation: Xinjiang Key Laboratory of Cardiovascular Disease Research, Urumqi 830054, Xinjiang Uygur Autonomous Region, China.
authors.forename: Bang-Dang
authors.initials: BD
authors.lastname: Chen
authors.affiliation: Xinjiang Key Laboratory of Cardiovascular Disease Research, Urumqi 830054, Xinjiang Uygur Autonomous Region, China.
authors.forename: Fen
authors.initials: F
authors.lastname: Liu
keywords: Adenovirus; Apoptosis; Cardiomyocytes; NF-κB pathway; R65 ribozyme