title: Esculetin induces apoptosis and inhibits adipogenesis in 3T3-L1 cells.
abstract: OBJECTIVE
  To determine the effects of esculetin, a plant phenolic compound with apoptotic activity in cancer cells, on 3T3-L1 adipocyte apoptosis and adipogenesis.
  
  
  METHODS
  3T3-L1 pre-confluent preadipocytes and lipid-filled adipocytes were incubated with esculetin (0 to 800 microM) for up to 48 hours. Viability was determined using the Cell Titer 96 Aqueous One Solution cell proliferation assay; apoptosis was quantified by measurement of single-stranded DNA. Post-confluent preadipocytes were incubated with esculetin for up to 6 days during maturation. Adipogenesis was quantified by measuring lipid content using Nile Red dye; cells were also stained with Oil Red O for visual confirmation of effects on lipid accumulation.
  
  
  RESULTS
  In mature adipocytes, esculetin caused a time- and dose-related increase in adipocyte apoptosis and a decrease in viability. Apoptosis was increased after only 6 hours by 400 and 800 microM esculetin (p < 0.05), and after 48 hours, as little as 50 microM esculetin increased apoptosis (p < 0.05). In preadipocytes, apoptosis was detectable only after 48 hours (p < 0.05) with 200 microM esculetin and higher concentrations. However, results of the cell viability assay indicated a reduction in preadipocyte number in a time- and dose-related manner, beginning as early as 6 hours with 400 and 800 microM esculetin (p < 0.05). Esculetin also inhibited adipogenesis of 3T3-L1 preadipocytes. Esculetin-mediated inhibition of adipocyte differentiation occurred during the early, intermediate, and late stages of the differentiation process. In addition, esculetin induced apoptosis during the late stage of differentiation.
  
  
  CONCLUSIONS
  These findings suggest that esculetin can alter fat cell number by direct effects on cell viability, adipogenesis, and apoptosis in 3T3-L1 cells.
authors.affiliation: Department of Animal and Dairy Science, University of Georgia, Athens, GA 30602-2771, USA.
authors.forename: Jeong-Yeh
authors.initials: JY
authors.lastname: Yang
authors.forename: Mary Anne
authors.initials: MA
authors.lastname: Della-Fera
authors.forename: Diane L
authors.initials: DL
authors.lastname: Hartzell
authors.forename: Cass
authors.initials: C
authors.lastname: Nelson-Dooley
authors.forename: Dorothy B
authors.initials: DB
authors.lastname: Hausman
authors.forename: Clifton A
authors.initials: CA
authors.lastname: Baile
mesh_terms: D041721:3T3-L1 Cells; D017667:Adipocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D050156:Adipogenesis / Q000187:drug effects*; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D002452:Cell Count; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D008055:Lipids / Q000032:analysis; D051379:Mice; D013997:Time Factors; D014468:Umbelliferones / Q000494:pharmacology*
chemical_list: D000975:Antioxidants; D008055:Lipids; D014468:Umbelliferones; C007628:esculetin