Suppression of Nuclear Factor-κB Activity in Kupffer Cells Protects Rat Liver Graft From Ischemia-Reperfusion Injury

Abstract

Objective The objective of this study was to investigate the protective effect and mechanisms of nuclear factor (NF)-κB 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 μg NF-κB decoy ODN before procurement. Following 2 hours of reperfusion, NF-κB binding activity was detected in isolated Kupffer cells (KCs) using electrophoretic mobility shift assays (EMSA). Tumor necrosis factor (TNF)-α 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-κB binding activity, TNF-α 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-κB activation following reperfusion plays an important role in IRI after liver transplantation. The decoy strategy showed an apparent effect to suppress NF-κB activation and inhibit production of downstream cytokines, thereby protecting liver grafts from IRI.