Cell-based screening assay for anti-inflammatory activity of bioactive compounds

Kees Meijer*, Roel J. Vonk, Marion G. Priebe, Han Roelofsen

*Corresponding author for this work

    Research output: Contribution to journalArticleAcademicpeer-review

    13 Citations (Scopus)

    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.

    Original languageEnglish
    Pages (from-to)158-164
    Number of pages7
    JournalFood Chemistry
    Volume166
    DOIs
    Publication statusPublished - 1-Jan-2015

    Keywords

    • Reporter assay
    • NF-kappa B
    • Anti-inflammatory
    • HEK293
    • Sulphoraphane
    • Lutein
    • SCFA
    • Butyrate
    • NF-KAPPA-B
    • CHAIN FATTY-ACIDS
    • ADIPOSE-TISSUE
    • TUMOR-SUPPRESSOR
    • GENE-EXPRESSION
    • PROPIONIC-ACID
    • NITRIC-OXIDE
    • SULFORAPHANE
    • LUTEIN
    • INFLAMMATION

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