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PLoS By Category | Recent PLoS Articles
Molecular Biology - Respiratory Medicine

Imaging Pulmonary NF-kappaB Activation and Therapeutic Effects of MLN120B and TDZD-8
Published: Thursday, September 22, 2011
Author: Dan Ansaldi et al.

by Dan Ansaldi, Eldad A. Hod, Fabio Stellari, Jae-Beom Kim, Ed Lim, Mark Roskey, Kevin P. Francis, Rajendra Singh, Ning Zhang

NF-?B activation is a critical signaling event in the inflammatory response and has been implicated in a number of pathological lung diseases. To enable the assessment of NF-?B activity in the lungs, we transfected a luciferase based NF-?B reporter into the lungs of mice or into Raw264.7 cells in culture. The transfected mice showed specific luciferase expression in the pulmonary tissues. Using these mouse models, we studied the kinetics of NF-?B activation following exposure to lipopolysaccharide (LPS). The Raw264.7 cells expressed a dose-dependent increase in luciferase following exposure to LPS and the NF-?B reporter mice expressed luciferase in the lungs following LPS challenge, establishing that bioluminescence imaging provides adequate sensitivity for tracking the NF-?B activation pathway. Interventions affecting the NF-?B pathway are promising clinical therapeutics, thus we further examined the effect of IKK-2 inhibition by MLN120B and glycogen synthase kinase 3 beta inhibition by TDZD-8 on NF-?B activation. Pre-treatment with either MLN120B or TDZD-8 attenuated NF-?B activation in the pulmonary tissues, which was accompanied with suppression of pro-inflammatory chemokine MIP-1ß and induction of anti-inflammatory cytokine IL-10. In summary, we have established an imaging based approach for non-invasive and longitudinal assessment of NF-?B activation and regulation during acute lung injury. This approach will potentiate further studies on NF-?B regulation under various inflammatory conditions.
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