Suppression of nuclear factor-kappaB activity in Kupffer cells protects rat liver graft from ischemia-reperfusion injury.
OBJECTIVE
The objective of this study was to investigate the protective effect and mechanisms of nuclear factor (NF)-kappaB decoy oligodeoxynucleotides (ODN) on rat liver grafts following ischemia-reperfusion injury (IRI).


METHODS
Animals were randomly divided into 3 groups (n = 8): control ischemia-reperfusion (IR) and decoy ODN groups; in the last cohort donor grafts were transfected with 120 microg NF-kappaB decoy ODN before procurement. Following 2 hours of reperfusion, NF-kappaB binding activity was detected in isolated Kupffer cells (KCs) using electrophoretic mobility shift assays (EMSA). Tumor necrosis factor (TNF)-alpha and interleukin (IL)-6 messenger RNA (mRNA) expressions were analyzed using reverse transcriptase polymerase chain reaction (RT-PCR) methods. Liver tissue and blood serum were collected for histopathologic examination and liver function test, respectively.


RESULTS
The NF-kappaB binding activity, TNF-alpha and IL-6 mRNA expression as well as serum ALT and total bilirubin levels were significantly increased compared with the control group following reperfusion (P < .01). A large number of hepatocytes showed degeneration and necrosis. However, these indices were significantly ameliorated among the decoy ODN group (P < .01) with preserved hepatic lobule architecture.


CONCLUSIONS
KCs NF-kappaB activation following reperfusion plays an important role in IRI after liver transplantation. The decoy strategy showed an apparent effect to suppress NF-kappaB activation and inhibit production of downstream cytokines, thereby protecting liver grafts from IRI.
Li
J D
JD
Department of Hepatobiliary Surgery, Affiliated Hospital of North Sichuan Medical College, Nanchong, Sichuan Province, China.
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D000410:Alanine Transaminase / Q000097:blood; D000818:Animals; D001219:Aspartate Aminotransferases / Q000097:blood; D017931:DNA Primers; D015850:Interleukin-6 / Q000235:genetics; D007728:Kupffer Cells / Q000502:physiology*; D016031:Liver Transplantation / Q000473:pathology / Q000502:physiology*; D008297:Male; D016328:NF-kappa B / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D009838:Oligodeoxyribonucleotides / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D015427:Reperfusion Injury / Q000517:prevention & control*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014184:Transplantation, Homologous / Q000502:physiology; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics
D017931:DNA Primers; D015850:Interleukin-6; D016328:NF-kappa B; D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D001219:Aspartate Aminotransferases; D000410:Alanine Transaminase