Suppression of nuclear factor-kappaB activity in Kupffer cells protects rat liver graft from ischemia-reperfusion injury.

摘要

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. CONCLUSION: 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.