NF-κB Decoy Oligodeoxynucleotides Inhibits Kupffer Cell Activation

Abstract

Objective: To investigate the inhibitory effects and the mechanism of NF-κB decoy oligodeoxynucleotides (ODN) on Kupffer cells (KCs) activation. Methods: KCs were isolated and randomly divided into three groups: (1) a control group; (2) an LPS stimu1ation group and (3) an NF-κB decoy group, in which the KCs were transduced with an NF-κB decoy ODN prior to LPS stimulation. Following 6 hours of LPS stimulation, the NF-κB activity was assayed by an electrophoretic mobility shift assay (EMSA). The CD80 mRNA expression in the KCs was detected by reverse transcription-polymerase chain reaction (RT-PCR), and the production of TNF-α and IL-6 in the supernatant was measured by an enzyme-linked immune sorbent assay (ELISA). Results: The NF-κB decoy ODN could efficiently inhibit KCs activation by LPS stimulus. The NF-κB activity was significantly decreased to 0.53 fold as compared with the LPS group. The CD80 mRNA expression, TNF-α production, and IL-6 level were significantly decreased to 0.46, 0.37, and 0.60 fold, respectively. Conclusion: The NF-κB decoy ODN could efficiently suppress transcription activity of NF-κB and inhibit co-stimulatory molecules and cytokines expression by KCs, which afford reliable experimental data for the in vivo application of NF-κB decoy ODN. Keywords: Cytokines, Kupffer cells, LPS, NF-κB, oligodeoxynucleotides, TNF-α.