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Development of in vitro and in vivo anti‐inflammatory assays for testing cyanobacterial marine natural products
Author(s) -
Villa Francisco A,
Gerwick Lena,
Gerwick William H
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.2_supplement.537
Subject(s) - in vivo , in vitro , reporter gene , innate immune system , chemistry , nitric oxide , zebrafish , green fluorescent protein , in vitro toxicology , ic50 , biochemistry , biology , microbiology and biotechnology , pharmacology , gene , gene expression , receptor , organic chemistry
Due to evolutionary selective pressure prokaryotes have developed a rich array of secondary metabolites, some of which may be inhibitory to the innate immune system and the inflammatory response in vertebrates. With this rationale in mind we have developed a nitric oxide (NO) assay and a GFP reporter assay to screen and characterize in vitro the anti‐inflammatory activities of compounds from a library of over 130 pure cyanobacterial marine natural products. Twelve compounds from this library exhibited anti‐inflammatory activity in the NO assay with IC50 values of 3–15 μg/mL. These 12 compounds were further tested in iNOS‐ and COX2‐GFP reporter assays to confirm these findings and to begin to elucidate their mechanism of action. We are in the process of developing a COX2‐GFP transgenic zebrafish to use as an in vivo model to measure the anti‐inflammatory activity of these compounds. Results from the reporter assay will be presented.