title: Cell-based screening assay for anti-inflammatory activity of bioactive compounds.
abstract: Excess dietary intake may induce metabolic inflammation which is associated with insulin resistance and cardiovascular disease. Recent evidence indicates that dietary bioactive compounds may diminish metabolic inflammation. To identify anti-inflammatory bioactives, we developed a screening assay using the human H293-NF-κB-RE-luc2P reporter cell line. Under optimised conditions we determined the anti-inflammatory activity of vegetables and purified bioactives, by monitoring their potency to inhibit TNF-α-induced NF-κB activity, as assessed by sensitive chemiluminescence detection in a 96-well assay format. Minced broccoli seedlings reduced NF-κB activity by 16%, while sulphoraphane, the dominant bioactive in broccoli seedlings, inhibited NF-κB activity with an IC₅₀ of 5.11 μmol/l. Short-chain fatty acids also reduced NF-κB activity in the order butyrate>propionate≫acetate with IC₅₀ of 51, 223, and 1300 μmol/l, respectively. The H293-NF-κB-RE-luc2P reporter cell line is a sensitive tool for rapid high-throughput screening for bioactives with anti-inflammatory activity.
authors: Center for Medical Biomics, University Medical Centre Groningen, Antonius Deusinglaan 1, 9713AV Groningen, The Netherlands. Electronic address: kornelis.meijer@umcg.nl.
authors: Kees
authors: K
authors: Meijer
authors: Center for Medical Biomics, University Medical Centre Groningen, Antonius Deusinglaan 1, 9713AV Groningen, The Netherlands.
authors: Roel J
authors: RJ
authors: Vonk
authors: Center for Medical Biomics, University Medical Centre Groningen, Antonius Deusinglaan 1, 9713AV Groningen, The Netherlands.
authors: Marion G
authors: MG
authors: Priebe
authors: Center for Medical Biomics, University Medical Centre Groningen, Antonius Deusinglaan 1, 9713AV Groningen, The Netherlands.
authors: Han
authors: H
authors: Roelofsen
mesh_terms: D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D057809:HEK293 Cells; D006801:Humans; D007249:Inflammation / Q000188:drug therapy*; D017879:Isothiocyanates / Q000737:chemistry*; D016328:NF-kappa B / Q000378:metabolism*; D013454:Sulfoxides; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D000893:Anti-Inflammatory Agents; D017879:Isothiocyanates; D016328:NF-kappa B; D013454:Sulfoxides; D014409:Tumor Necrosis Factor-alpha; C016766:sulforaphane
keywords: Anti-inflammatory; Butyrate; HEK293; Lutein; NF-κB; Reporter assay; SCFA; Sulphoraphane