Abstract
NF-kB plays a central role in immunity, inflammation, development, cell survival and has been indicated under a number of pathological conditions of lung disease, including asthma, chronic bronchitis, and chronic obstructive pulmonary disease. In this study, we assessed the in vivo activation of NF-kB signaling in lung tissue using a bioluminescence imaging system (IVIS) to monitor activation of an NF-kB promoter in response to lipopolysaccharide stimulation. A plasmid contained responsive elements of NF-kB and luciferase as a reporter gene has been delivered intravenously in nude mice at the concentration of 40 ug per mouse using in vivo-jetPEI™ from Polyplus as a transfectant agent. One week after DNA delivery the transient transgenic mice had been imaged in order to check the baseline activation of the NF-kB pathway. The day after, the mice have been treated with LPS 15 ug per mouse intratracheally and the lungs imaged using bioluminescence (BLI) at 2, 4, 7 and 24 hs. The ability of the IKK2 inhibitor MLN120B orally administered at the dose of 300 mg/kg to counteract NF-kB activation has been evaluated. The maximum peak of NF-kB activation was reached at 4 hs with 7-10 folds of induction in comparison to the saline group and at 24 hs the signal dropped down at basal level. In the group treated with MLN120B was observed a 50% inhibition of LPS-induced NF-kB stimulation, an effect that was in good agreement with the inhibition of p65 nuclear translocation evaluated ex vivo in lung homogenates.
In this experiment we showed that is feasible to monitor NF-kB activation in vivo in lung tissue in a non-invasive way by BLI and the pharmacological response as well.
- © 2011 ERS
