These authors equally contributed to this work.
Adipocyte reporter assays: Application for identification of anti-inflammatory and antioxidant properties of mangosteen xanthones
Article first published online: 3 SEP 2013
© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Molecular Nutrition & Food Research
Volume 58, Issue 2, pages 239–247, February 2014
How to Cite
Shen, Q., Chitchumroonchokchai, C., Thomas, J. L., Gushchina, L. V., DiSilvestro, D., Failla, M. L. and Ziouzenkova, O. (2014), Adipocyte reporter assays: Application for identification of anti-inflammatory and antioxidant properties of mangosteen xanthones. Mol. Nutr. Food Res., 58: 239–247. doi: 10.1002/mnfr.201300181
- Issue published online: 3 FEB 2014
- Article first published online: 3 SEP 2013
- Manuscript Accepted: 26 JUN 2013
- Manuscript Revised: 14 JUN 2013
- Manuscript Received: 11 MAR 2013
- Food Innovation Center, Office for International Affairs and Center for Advanced Functional Foods Research and Entrepreneurship at OSU
- National Center for Research Resources. Grant Number: UL1RR025755
- Office of the Director, National Institutes of Health (OD)
- NIH Roadmap for Medical Research
- Nuclear receptors;
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.
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.