13 March 2012 Mechanistic study of macrophage activation by LPS stimulation using fluorescence imaging techinques
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Abstract
Lipopolysaccharide (LPS), a structural component of the outer membrane of gram negative bacteria, has been suggested that stimulates macrophages secrete a wide variety of inflammatory mediators, such as nitric oxide (NO). However, the cellular mechanisms of NO generation in macrophage by LPS stimulation are not well known. In this study, LPS stimulated NO generation in macrophage was determined by measuring fluorescence changes with a NO specific probe DAF-FM DA. Using the fluorescence resonance energy transfer (FRET) techniques, we found an increase of protein kinase C (PKC) activation was dynamically monitored in macrophages treated with LPS. Nuclear factor kappa B (NF-κB) translocated from the cytoplasm to the nucleus in macrophage was measured by confocal laser scanning microscopy. Moreover, the PKC inhibitor GÖ6983 inhibited LPS-stimulated NF-κB activation and NO production. These results indicated that LPS stimulated NF-κB mediated NO production by activating PKC.
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Cuixia Lu, Feifan Zhou, Wei R. Chen, Da Xing, "Mechanistic study of macrophage activation by LPS stimulation using fluorescence imaging techinques", Proc. SPIE 8329, Tenth International Conference on Photonics and Imaging in Biology and Medicine (PIBM 2011), 83290N (13 March 2012); doi: 10.1117/12.919009; https://doi.org/10.1117/12.919009
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