Gene transduction of an active mutant of akt exerts cytoprotection and reduces graft injury after liver transplantation.
Akt is expected to be an effective target for the treatment of ischemia-reperfusion injury (I/R) due to its anti-apoptotic properties and its ability to activate the endothelial nitric oxide synthase (eNOS) enzyme. Therefore, this study was aimed to determine the efficacy of an active mutant of Akt (myr-Akt) to decrease I/R injury in a model of orthotopic liver transplantation in pigs. In addition, we analyzed the contribution of nitric oxide in the Akt-mediated effects by using an eNOS mutant (S1179DeNOS) that mimics the phosphorylation promoted by Akt in the eNOS sequence. Donors were treated with adenoviruses codifying for myr-Akt, S1179DeNOS or beta-galactosidase 24 h before liver harvesting. Then, liver grafts were orthotopically transplanted into their corresponding recipients. Levels of transaminases and lactate dehydrogenase (LDH) increased in all recipients after 24 h of transplant. However, transaminases and LDH levels were significantly lower in the myr-Akt group compared with vehicle. The percentage of apoptotic cells and the amount of activated-caspase 3 protein were also markedly reduced in myr-Akt-treated grafts after 4 days of liver transplant compared with vehicle and S1179DeNOS groups. In conclusion, myr-Akt gene therapy effectively exerts cytoprotection against hepatic I/R injury regardless of the Akt-dependent eNOS activation.
Morales-Ruiz
M
M
Department of Biochemistry and Molelular Genetics, Hospital Clinic, IDIBAPS, University of Barcelona and IRSIN, Spain.
Fondevila
C
C
Muñoz-Luque
J
J
Tugues
S
S
Rodríguez-Laiz
G
G
Cejudo-Martín
P
P
Romero
J M
JM
Navasa
M
M
Fuster
J
J
Arroyo
V
V
Sessa
W C
WC
García-Valdecasas
J C
JC
Jiménez
W
W
D000818:Animals; D001011:Aorta; D002417:Cattle; D002460:Cell Line; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology* / Q000502:physiology; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology*; D022781:Hepatocytes / Q000166:cytology / Q000502:physiology; D006801:Humans; D007668:Kidney; D009154:Mutation; D051058:Oncogene Protein v-akt / Q000235:genetics / Q000502:physiology*; D013552:Swine
D051058:Oncogene Protein v-akt