Analysis of in vivo nuclear factor-kappaB activation during liver inflammation in mice: prevention by catalase delivery.
Nuclear factor-kappaB (NF-kappaB) is a transcription factor that plays crucial roles in inflammation, immunity, cell proliferation, and apoptosis. Until now, there have been few studies of NF-kappaB activation in whole animals because of experimental difficulties. Here, we show that mice receiving a simple injection of plasmid vectors can be used to examine NF-kappaB activation in the liver. Two plasmid vectors, pNF-kappaB-Luc (firefly luciferase gene) and pRL-SV40 (Renilla reniformis luciferase gene), were injected into the tail vein of mice by the hydrodynamics-based procedure, an established method of gene transfer to mouse liver. Then, the ratio of the firefly and R. reniformis luciferase activities (F/R) was used as an indicator of the NF-kappaB activity in the liver. Injection of thioacetamide or lipopolysaccharide plus d-galactosamine increased the F/R ratio in the liver, and this was significantly (P<0.001) inhibited by an intravenous injection of catalase derivatives targeting liver nonparenchymal cells. Imaging the firefly luciferase expression in live mice clearly demonstrated that the catalase derivatives efficiently prevented the NF-kappaB-mediated expression of the firefly luciferase gene. Plasma transaminases and the survival rate of mice supported the findings obtained by the luminescence-based analyses. Thus, this method, which requires no genetic recombination techniques, is highly sensitive to the activation of NF-kappaB and allows us to continuously examine the activation in live animals. In conclusion, this novel, simple, and sensitive method can be used not only for analyzing the NF-kappaB activation in the organ under different inflammatory conditions but also for screening drug candidates for the prevention of liver inflammation.
Hyoudou
Kenji
K
Department of Drug Delivery Research, Graduate School of Pharmaceutical Sciences, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan.
Nishikawa
Makiya
M
Kobayashi
Yuki
Y
Kuramoto
Yukari
Y
Yamashita
Fumiyoshi
F
Hashida
Mitsuru
M
D000818:Animals; D002374:Catalase / Q000008:administration & dosage / Q000494:pharmacology*; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000008:administration & dosage; D007249:Inflammation / Q000188:drug therapy / Q000378:metabolism*; D008099:Liver / Q000187:drug effects / Q000473:pathology; D008107:Liver Diseases / Q000473:pathology*; D008156:Luciferases / Q000235:genetics; D008163:Luminescent Measurements / Q000379:methods*; D008297:Male; D008722:Methods; D051379:Mice; D008815:Mice, Inbred Strains; D016328:NF-kappa B / Q000032:analysis / Q000378:metabolism*; D015996:Survival Rate
D016328:NF-kappa B; D002374:Catalase; D008156:Luciferases