Selective inactivation of NF-κB in the liver using NF-κB decoy suppresses CCl4-induced liver injury and fibrosis

Sustained hepatic inflammation induced by various causes can lead to liver fibrosis. Transcription factor NF-kappa B is important in regulating inflammatory responses, especially in macrophages. We presently investigated whether an NF-kappa B decoy, a synthetic oligodeoxynucleotide (ODN) imitating the NF-kappa B binding site, inhibited the inflammatory response after CCl4 intoxication to prevent CCl4-induced hepatic injury and fibrosis. The NF-kappa B decoy was introduced into livers by injecting the spleens of mice, using a hemagglutinating virus of Japan (HVJ)-liposome method. ODN was transferred mainly to macrophages in normal or fibrotic livers. Increases in serum transaminases and production of inflammatory cytokines after a single challenge with CCl4 were inhibited by the NF-kappa B decoy, which suppressed nuclear translocation of NF-kappa B in liver macrophages. Liver fibrosis induced by CCl4 administration for 8 wk was suppressed by the NF-kappa B decoy, accompanied by diminished mRNA expression for transforming growth factor (TGF)-beta, procollagen type 1 alpha 1, and alpha-smooth muscle actin (SMA). In vitro, isolated liver macrophages showed increased DNA binding activity of NF-kappa B and inflammatory cytokine production after hydrogen peroxide treatment; both increases were inhibited significantly by the NF-kappa B decoy. In contrast, NF-kappa B decoy transferred to isolated hepatic stellate cells (HSC) had no effect on their morphological activation or alpha-SMA expression, although the decoy accelerated tumor necrosis factor (TNF)-alpha-induced apoptosis in activated HSC. The effect of NF-kappa B decoy suppressing fibrosis probably results mainly from anti-inflammatory effects on liver macrophages, with a possible minor contribution from its direct proapoptotic effect on activated HSC.