Transplantation · 2008
A molecular approach to apoptosis in the human heart during brain death.
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Pérez López Silvia, Vázquez Moreno Natalia, Escudero Augusto Dolores, Astudillo González Aurora, Alvarez Menéndez Francisco, Goyache Goñi Félix, Otero Hernández Jesús
Аннотация
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.
CONCLUSION: Our findings suggest the possibility of using antiapoptosis agents to prevent cardiac injury and improve posttransplant behavior.
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.
CONCLUSION: Our findings suggest the possibility of using antiapoptosis agents to prevent cardiac injury and improve posttransplant behavior.
Предметные рубрики MeSH
AdultAgedAged, 80 and overApoptosisBrain DeathCaspase 3Cytochromes cDNA PrimersFemaleGene Expression RegulationHeartHeart Valve Prosthesis ImplantationHeart VentriclesHumansHypoxia-Inducible Factor 1, alpha SubunitMaleMiddle AgedMitral Valve ProlapseModels, BiologicalMyocardiumProto-Oncogene Proteins c-bcl-2Ventricular Functionbcl-2-Associated X Proteinfas Receptor