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Monitoring of Endotoxin-Induced Pulmonary Inflammation In Vivo in NF-kB Luciferase Transgenic Mice

Rationale: NF-kappaB is necessary for coordinating the activation of many genes involved in inflammatory responses in the lungs. We explored the use of NF-kB-luciferase BALB/c transgenic mice to evaluate the relationship between inhaled endotoxin and the activation of NF-kB signals in lungs in vivo. Methods: Xenogen NF-kB BALB/c mice with 3 luciferase reporter elements were exposed to N. meningitidis endotoxin (0-300, 000 EU/mouse) and saline (controls) by nasal aspiration. Before exposure and after certain time points (4, 8hr) luciferin was injected intraperitoneally and mice were imaged with an ultrasensitive CCD camera (IVIS-100, Xenogen). After imaging the thoracic region, mice were euthanized and lungs were rapidly excised and imaged. The oxyluciferin signals from the thoracic region and lungs were quantified using Living Image Software. Results: The imaging of exposed transgenic NF-kB-luc BALB/c mice demonstrated a quantifiable endotoxin induced signal in the thoracic region in vivo over controls at doses of endotoxin above 3, 000 EU/mouse. The signal in the lungs of exposed mice imaged ex vivo showed high values (5 to 12-fold increases) over control and provided documentation of NF-kB activation in the lung. Conclusions: Markers of lung inflammation from bronchoalveolar lavage such as neutrophil recruitment and cytokine concentrations were more sensitive biomarkers than imaging oxyluciferin photonic yield in vivo. A lower dose of inhaled endotoxin was capable of inducing measurable lung luminescence compared with controls when lungs were imaged ex vivo. This study provides new data regarding the sensitivity of NF-kB-luc transgenic mice and their utility for in vivo monitoring of lung inflammation. Funded By: NIH P30 ES05605-15

NF-kappaB; lungs; transgenic mice; inflammatory responses

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