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Adipocyte reporter assays: Application for identification of anti‐inflammatory and antioxidant properties of mangosteen xanthones
Author(s) -
Shen Qiwen,
Chitchumroonchokchai Chureeporn,
Thomas Jaisa L.,
Gushchina Liubov V.,
DiSilvestro David,
Failla Mark L.,
Ziouzenkova Ouliana
Publication year - 2014
Publication title -
molecular nutrition and food research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.495
H-Index - 131
eISSN - 1613-4133
pISSN - 1613-4125
DOI - 10.1002/mnfr.201300181
Subject(s) - adipogenesis , peroxisome proliferator activated receptor gamma , chemistry , inflammation , adipocyte , nutraceutical , peroxisome proliferator activated receptor , antioxidant , pharmacology , biochemistry , adipose tissue , medicine , biology , receptor
Scope Three fluorescence biosensors were developed based on a 3T3‐L1 preadipocyte line that stably expressed Nfkb ‐RE/GFP, Fabp4 ‐P/CFP, and Nrf2 ‐P/YFP fluorescent reporters. We hypothesized that nutraceuticals’ inflammatory, adipogenic, and antioxidant status will be identified based on the change in fluorescence in reporter adipocytes. We validated these assays with activators of NFκB, FABP4‐regulating peroxisome proliferator activated receptor gamma, NFR2 and, thereafter, tested known and unknown properties of mangostines (MGs), the xanthone metabolites in mangosteen fruit. Methods and results We validated inflammatory and adipogenic properties of α‐MG using an Nfkb‐ RE/GFP biosensor assay. Next, we identified unique properties of γ‐MG, a minor MG xanthone. γ‐MG suppressed adipogenesis and expression of adiponectin, but inhibited the Nfkb ‐RE/GFP reporter and secretion of inflammatory monocyte chemotactic protein 1 as compared to the control adipocytes. We found that the inhibition of adipogenesis and Nfkb ‐mediated inflammation depends on a dose‐dependent reduction of Nrf2 promoter activity by α‐MG. The Nrf2 inhibition resulted in the reduced Pparg expression. α‐MG did not directly influence Pparg activity in Fabp4 ‐P/CFP adipocytes. Conclusion α‐MG‐mediated antioxidant response via Nrf2 is a mechanism preventing adipogenesis and inflammation in adipocytes. Combined application of high‐throughput biosensors could provide an effective platform for the identification of nutraceuticals and the mechanism of their actions in adipocytes and, potentially, in obese patients.