Prolonged survival of mice with acute liver failure with transplantation of monkey hepatocytes cultured with an antiapoptotic pentapeptide V5.
BACKGROUND
Because hepatocyte transplantation has been considered to be an attractive method to treat acute liver failure (ALF), efficient recovery of hepatocytes and maintenance of differentiated hepatocyte functions is of extreme importance. We here report the usefulness of an antiapoptotic pentapeptide V5, composed of Val-Pro-Met-Leu-Lys, in the monkey hepatocyte cultures.


METHODS
We evaluated albumin production, metabolizing abilities of ammonia, lidocaine, and diazepam of monkey hepatocytes cultured with V5. The protein expression of apoptosis-associated molecules was analyzed using power blot analysis. An unwoven cloth inoculated with V5-treated monkey hepatocytes was transplanted on the surface of the spleen of both SCID mice and Balb/c mice suffering from ALF induced by 90% hepatectomy.


RESULTS
When 100 microM V5 was utilized, ammonia-, lidocaine- and diazepam- metabolizing capacities and albumin production ability were significantly increased in V5-treated monkey hepatocytes. Such hepatocytes showed decreased Annexin V binding and increased the expression of anti-apoptotic and/or cytoprotective molecules, including Ku70, NF-kappaB, IKAP, hILP/XIAP, IkappaB, and CAS. Transplantation of the cloth containing the monkey hepatocytes significantly improved blood levels of glucose and ammonia and encephalopathy score and prolonged the survival of the mice with ALF.


CONCLUSIONS
The present work clearly demonstrates the usefulness of V5 for maintaining the functions of monkey hepatocytes in tissue culture.
Department of Surgery, Okayama University Graduate School of Medicine and Dentistry, Okayama, Japan.
Kimiaki
K
Tanaka
Naoya
N
Kobayashi
Alejandro Soto
AS
Gutierrez
Jorge David
JD
Rivas-Carrillo
Nalu
N
Navarro-Alvarez
Yong
Y
Chen
Michiki
M
Narushima
Atsushi
A
Miki
Teru
T
Okitsu
Hirofumi
H
Noguchi
Noriaki
N
Tanaka
D000641:Ammonia / Q000378:metabolism; D000818:Animals; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D003975:Diazepam / Q000378:metabolism; D000882:Haplorhini; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism / Q000637:transplantation*; D008012:Lidocaine / Q000378:metabolism; D008099:Liver / Q000166:cytology; D017114:Liver Failure, Acute / Q000601:surgery*; D008115:Liver Regeneration; D051379:Mice; D008807:Mice, Inbred BALB C; D016513:Mice, SCID; D008930:Mitochondria, Liver / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D009842:Oligopeptides / Q000494:pharmacology*; D012709:Serum Albumin / Q000096:biosynthesis / Q000235:genetics; D013154:Spleen / Q000166:cytology / Q000601:surgery; D016019:Survival Analysis; D014183:Transplantation, Heterologous; D016896:Treatment Outcome
D009842:Oligopeptides; D012709:Serum Albumin; C509183:Val-Pro-Met-Leu-Lys; D000641:Ammonia; D008012:Lidocaine; D003975:Diazepam