title: Adipocyte reporter assays: application for identification of anti-inflammatory and antioxidant properties of mangosteen xanthones.
abstract: METHODS
  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.
  
  
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Human Sciences, The Ohio State University, Columbus, OH, USA.
authors.forename: Qiwen
authors.initials: Q
authors.lastname: Shen
authors.forename: Chureeporn
authors.initials: C
authors.lastname: Chitchumroonchokchai
authors.forename: Jaisa L
authors.initials: JL
authors.lastname: Thomas
authors.forename: Liubov V
authors.initials: LV
authors.lastname: Gushchina
authors.forename: David
authors.initials: D
authors.lastname: Disilvestro
authors.forename: Mark L
authors.initials: ML
authors.lastname: Failla
authors.forename: Ouliana
authors.initials: O
authors.lastname: Ziouzenkova
mesh_terms: D041721:3T3-L1 Cells; D017667:Adipocytes / Q000187:drug effects* / Q000378:metabolism; D050156:Adipogenesis / Q000187:drug effects; D052242:Adiponectin / Q000235:genetics / Q000378:metabolism; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D000975:Antioxidants / Q000494:pharmacology*; D015374:Biosensing Techniques; D002454:Cell Differentiation / Q000187:drug effects; D050556:Fatty Acid-Binding Proteins / Q000235:genetics / Q000378:metabolism; D005638:Fruit / Q000737:chemistry; D029762:Garcinia mangostana / Q000737:chemistry*; D016086:Lentivirus; D051379:Mice; D051267:NF-E2-Related Factor 2 / Q000235:genetics / Q000378:metabolism; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism; D047495:PPAR gamma / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D044004:Xanthones / Q000494:pharmacology*
chemical_list: D052242:Adiponectin; D000893:Anti-Inflammatory Agents; D000975:Antioxidants; C493411:Fabp4 protein, mouse; D050556:Fatty Acid-Binding Proteins; D051267:NF-E2-Related Factor 2; D016328:NF-kappa B; C495636:Nfe2l2 protein, mouse; D047495:PPAR gamma; D044004:Xanthones
keywords: Inflammation; Mangosteen; Nrf2; Nuclear receptors; Obesity
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