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1.
Biotechnol Lett ; 35(1): 11-20, 2013 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-22983717

RESUMEN

Using an in vitro cell-based assay in a flow-design, we have applied activity-guided screening to search for new bioactive compounds isolated from microorganisms. A first assay employs the stable expression of nuclear factor kappa B (NF-κB) while a second assay utilizes the glucocorticoid receptor (GR) coupled to green fluorescent protein. A specialized assay was implemented for both the translocation of NF-κB and to inhibit the translocation of cytokine-mediated NF-κB. In addition, we developed in a wide palette of cell lines used for a highly specialized GR-translocation assay to detect anti-inflammatory effects. This approach demonstrates the straight-forward combination of cell-based assays arranged with an automated fluorescence microscope. This allows for the direct sorting of extracts which are acting in a pharmaceutically interesting way. Initial results using this technique have led to the detection of new anti-inflammatory steroids from bacterial crude extracts.


Asunto(s)
Antiinflamatorios/farmacología , Bioensayo/métodos , Productos Biológicos/farmacología , Evaluación Preclínica de Medicamentos/métodos , Hidroxiesteroides/farmacología , Receptores de Glucocorticoides/metabolismo , Animales , Antiinflamatorios/metabolismo , Productos Biológicos/metabolismo , Células CHO , Línea Celular , Cricetinae , Cricetulus , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Hidroxiesteroides/metabolismo , Ratones , FN-kappa B/genética , FN-kappa B/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Receptores de Glucocorticoides/genética , Transfección
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