A molecular approach to apoptosis in the human heart during brain death.
BACKGROUND
Brain death induces changes in tissues and organs destined for transplant at the cell, molecular, and endocrine level including cell death through apoptosis. This study was designed to examine apoptotic damage in cardiac tissue obtained from brain dead donors.


METHODS
Fifty tissue specimens from the left ventricles of individual donors were processed to evaluate changes in the expression levels of five genes involved in apoptosis (BAX, BCL2, CASPASE 3, CYTOCHROME C, and FAS) using the real time-polymerase chain reaction technique. Expression levels were quantified by the relative standard method and results normalized to the levels recorded for the endogenous control peptidylprolyl isomerase A. The HIF1alpha gene was also determined to check for the possibility of hypoxic damage. Control ventricular tissue specimens were obtained from patients undergoing mitral valve replacement.


RESULTS
Using a mixed linear model it was determined that the sample type (donor vs. control patient) significantly affected (P<0.0001) expression levels of the genes examined reflected by their Ct values. Three of the genes (BAX, CASPASE 3, and FAS) showed significantly higher (Student's t test, P<0.05) expression levels (4.89-, 7.85-, and 12.14-fold endogenous control values, respectively) in donors compared with control patients (2.31-, 2.64-, and 3.57-fold endogenous control values, respectively) indicating the activation of apoptosis during brain death.


CONCLUSIONS
Our findings suggest the possibility of using antiapoptosis agents to prevent cardiac injury and improve posttransplant behavior.
Pérez López
Silvia
S
Unidad de Coordinación de Trasplantes y Terapia Celular, Hospital Universitario Central de Asturias, Oviedo, Spain. silvia.perez@sespa.princast.es
Vázquez Moreno
Natalia
N
Escudero Augusto
Dolores
D
Astudillo González
Aurora
A
Alvarez Menéndez
Francisco
F
Goyache Goñi
Félix
F
Otero Hernández
Jesús
J
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D017209:Apoptosis / Q000502:physiology*; D001926:Brain Death; D053148:Caspase 3 / Q000235:genetics; D045304:Cytochromes c / Q000235:genetics; D017931:DNA Primers; D005260:Female; D005786:Gene Expression Regulation; D006321:Heart / Q000503:physiopathology*; D019918:Heart Valve Prosthesis Implantation; D006352:Heart Ventricles / Q000503:physiopathology; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics; D008297:Male; D008875:Middle Aged; D008945:Mitral Valve Prolapse / Q000601:surgery; D008954:Models, Biological; D009206:Myocardium / Q000201:enzymology / Q000473:pathology*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics; D016276:Ventricular Function; D051028:bcl-2-Associated X Protein / Q000235:genetics; D019014:fas Receptor / Q000235:genetics
D017931:DNA Primers; C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; D019014:fas Receptor; D045304:Cytochromes c; D053148:Caspase 3