title: Bcl-2-mediated inhibition of apoptosis in rat cardiac allografts worsens development of graft coronary artery disease.
abstract: BACKGROUND
  We hypothesized that adenovirally mediated Bcl-2 transfection of donor hearts would reduce the apoptosis that occurs during acute rejection while worsening the development of chronic graft coronary artery disease (GCAD).
  
  
  METHODS
  PVG donor hearts were treated with either AdvBcl-2 or AdvNull virus before heterotopic transplantation into ACI rats. Bcl-2 expression was assessed on post-operative day 4 (POD) 4 by western blot. Apoptosis was measured using (99m)Technetium-bound-annexin V imaging and caspase 3 activity assay. Allograft survival was determined in a separate cohort of animals. Long-term-treated animals were then assessed for measures of GCAD on POD 90.
  
  
  RESULTS
  Western blot analysis showed upregulation of Bcl-2 expression in AdvBcl-2-treated hearts. (99m)Tc-annexin V images demonstrated decreased uptake in the AdvBcl-2 group (1.41 +/- 0.33% vs 1.94 +/- 0.37%, p = 0.026). Caspase 3 activity was also significantly lower in this treatment group (0.112 +/- 0.032 vs 0.204 +/- 0.096, p = 0.049). Allograft survival was similar in both groups, respectively (7.7 +/- 1.2 vs 6.8 +/- 1.5 days, p = 0.340). GCAD, as determined by percent luminal narrowing (5.9 +/- 6.1% vs 1.6 +/- 1.5%, p = 0.039), intima-to-media ratio (5.1 +/- 5.1% vs 1.5 +/- 1.7%, p = 0.040) and percent of affected vessels (15.1 +/- 9.9% vs 5.3 +/- 4.4%, p = 0.009), was higher for the AdvBcl-2 group.
  
  
  CONCLUSIONS
  Treatment of cardiac allografts with AdvBcl-2 resulted in a reduction of apoptosis that did not significantly improve short-term graft survival, but worsened chronic GCAD.
authors.affiliation: Department of Cardiothoracic Surgery, Stanford University School of Medicine, Stanford, California, USA.
authors.forename: Murray H
authors.initials: MH
authors.lastname: Kown
authors.forename: Douglas N
authors.initials: DN
authors.lastname: Miniati
authors.forename: Christina L
authors.initials: CL
authors.lastname: Jahncke
authors.forename: Maarten A
authors.initials: MA
authors.lastname: Lijkwan
authors.forename: Seiichiro
authors.initials: S
authors.lastname: Murata
authors.forename: Mark L
authors.initials: ML
authors.lastname: Koransky
authors.forename: Francis G
authors.initials: FG
authors.lastname: Blankenberg
authors.forename: H William
authors.initials: HW
authors.lastname: Strauss
authors.forename: Robert C
authors.initials: RC
authors.lastname: Robbins
mesh_terms: D000818:Animals; D017304:Annexin A5 / Q000378:metabolism; D017209:Apoptosis; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D003327:Coronary Disease / Q000209:etiology*; D015870:Gene Expression; D019254:Genes, bcl-2; D016027:Heart Transplantation; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D016019:Survival Analysis; D014162:Transfection; D014184:Transplantation, Homologous
chemical_list: D017304:Annexin A5; C505410:Casp3 protein, rat; D053148:Caspase 3; D020169:Caspases