title: 4β-Hydroxywithanolide E isolated from Physalis pruinosa calyx decreases inflammatory responses by inhibiting the NF-κB signaling in diabetic mouse adipose tissue.
abstract: BACKGROUND
  Chronic inflammation in adipose tissue together with obesity induces insulin resistance. Inhibitors of chronic inflammation in adipose tissue can be a potent candidate for the treatment of diabetes; however, only a few compounds have been discovered so far. The objective of this study was to find a novel inhibitor that can suppress the inflammatory response in adipose tissue and to elucidate the intracellular signaling mechanisms of the compound.
  
  
  METHODS
  To find the active compounds, we established an assay system to evaluate the inhibition of induced MCP-1 production in adipocyte/macrophage coculture in a plant extract library. The active compound was isolated by performing high-performance liquid chromatography (HPLC) and was determined as 4β-hydroxywithanolide E (4βHWE) by nuclear magnetic resonance (NMR) and mass spectroscopy (MS) spectral analyses. The effect of 4βHWE on inflammation in adipose tissue was assessed with adipocyte culture and db/db mice.
  
  
  RESULTS
  During the screening process, Physalis pruinosa calyx extract was found to inhibit production of MCP-1 in coculture strongly. 4βHWE belongs to the withanolide family of compounds, and it has the strongest MCP-1 production inhibitory effect and lowest toxicity than any other withanolides in coculture. Its anti-inflammatory effect was partially dependent on the attenuation of NF-κB signaling in adipocyte. Moreover, in vivo experiments showed that the oral administration of 4βHWE to db/db mice resulted in the inhibition of macrophage invasion and cytokine expression in adipose tissue after 2 weeks of treatment; improved the plasma adiponectin, non-esterified fatty acids and MCP-1 concentrations; and increased glucose tolerance after 3 to 4 weeks of treatment.
  
  
  CONCLUSIONS
  These results suggest that 4βHWE has anti-inflammatory effect via inhibition of NF-κB activation in adipocyte. Moreover, the attenuation of inflammation in adipocyte has an effect on the inhibition of macrophage accumulation in obese adipose tissue. Consequently, 4βHWE improves impaired glucose tolerance. Thus, 4βHWE is a useful natural anti-inflammatory compound to attenuate progression of diabetes and obesity.
authors.affiliation: Frontier Research Laboratories, Institute for Innovation, Ajinomoto Co., Inc., Kawasaki-Ku, Kawasaki, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Takimoto
authors.affiliation: Frontier Research Laboratories, Institute for Innovation, Ajinomoto Co., Inc., Kawasaki-Ku, Kawasaki, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Kanbayashi
authors.affiliation: Frontier Research Laboratories, Institute for Innovation, Ajinomoto Co., Inc., Kawasaki-Ku, Kawasaki, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Toyoda
authors.affiliation: Frontier Research Laboratories, Institute for Innovation, Ajinomoto Co., Inc., Kawasaki-Ku, Kawasaki, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Adachi
authors.affiliation: Frontier Research Laboratories, Institute for Innovation, Ajinomoto Co., Inc., Kawasaki-Ku, Kawasaki, Japan.
authors.forename: C
authors.initials: C
authors.lastname: Furuta
authors.affiliation: Frontier Research Laboratories, Institute for Innovation, Ajinomoto Co., Inc., Kawasaki-Ku, Kawasaki, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Suzuki
authors.affiliation: Frontier Research Laboratories, Institute for Innovation, Ajinomoto Co., Inc., Kawasaki-Ku, Kawasaki, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Miwa
authors.affiliation: Frontier Research Laboratories, Institute for Innovation, Ajinomoto Co., Inc., Kawasaki-Ku, Kawasaki, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Bannai
mesh_terms: D041721:3T3-L1 Cells; D017667:Adipocytes / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D000273:Adipose Tissue / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D000818:Animals; D018932:Chemokine CCL2 / Q000037:antagonists & inhibitors* / Q000096:biosynthesis; D002851:Chromatography, High Pressure Liquid; D018920:Coculture Techniques; D005947:Glucose / Q000378:metabolism; D015151:Immunoblotting; D007249:Inflammation / Q000188:drug therapy*; D007333:Insulin Resistance; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D013058:Mass Spectrometry; D051379:Mice; D016688:Mice, Inbred NOD / Q000378:metabolism; D016328:NF-kappa B / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D019906:Nuclear Magnetic Resonance, Biomolecular; D032321:Physalis / Q000737:chemistry*; D008517:Phytotherapy; D010936:Plant Extracts / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects*; D054358:Withanolides / Q000302:isolation & purification / Q000494:pharmacology*
chemical_list: C554450:4-hydroxywithanolide E; C491768:Ccl2 protein, mouse; D018932:Chemokine CCL2; D016328:NF-kappa B; D010936:Plant Extracts; D054358:Withanolides; D005947:Glucose