title: Effects of sterculic acid on stearoyl-CoA desaturase in differentiating 3T3-L1 adipocytes.
abstract: The effects of sterculic acid on cell size, adiposity, and fatty acid composition of differentiating 3T3-L1 adipocytes are correlated with stearoyl-CoA desaturase (SCD) expression (mRNA and protein levels) and enzyme activity. Fluorescence-activated cell scanning (FACS) analysis showed that adipocytes differentiated with methylisobutylxanthine, dexamethasone, and insulin (MDI) plus 100 microM sterculic acid comprised a population of predominantly large cells with reduced adiposity compared to MDI-treated cells. Although both groups had similar amounts of total fat, their fatty acid profiles were strikingly different: MDI-treated cells had high levels of the unsaturated palmitoleic (Delta(9)-16:1) and oleic (Delta(9)-18:1) acids, whereas the cells cultured with MDI plus sterculic acid accumulated palmitic (16:0) and stearic (18:0) acids together with a marked reduction in Delta(9)-16:1. Although the cells treated with MDI plus sterculic acid had similar levels of scd1 and scd2 mRNAs and antibody-detectable SCD protein as the MDI-treated cells, the SCD enzyme activity was inhibited more than 90%. The accumulation of 16:0 and 18:0, together with normal levels of fatty acid synthase (FAS) and aP2 mRNAs, shows that de novo synthesis and elongation of fatty acids, as well as cell differentiation, were not affected by sterculic acid. Because of the increase in cell size in the sterculic acid-treated cells, the insulin-stimulated 2-deoxyglucose (2-DOG) uptake was determined. Compared to MDI-treated cells, the 2-DOG uptake in the cells treated with sterculic acid was not affected. These results indicate that sterculic acid directly inhibits SCD activity, possibly by a turnover-dependent reaction, without affecting the processes required for adipocyte differentiation, scd gene expression or SCD protein translation.
authors.affiliation: Department of Biochemistry, College of Agricultural and Life Sciences, University of Wisconsin, Room 415B, 433 Babcock Drive, Madison WI 53706, USA.
authors.forename: F Enrique
authors.initials: FE
authors.lastname: Gomez
authors.forename: Dale E
authors.initials: DE
authors.lastname: Bauman
authors.forename: James M
authors.initials: JM
authors.lastname: Ntambi
authors.forename: Brian G
authors.initials: BG
authors.lastname: Fox
mesh_terms: D016475:3T3 Cells; D017667:Adipocytes / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology*; D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D002454:Cell Differentiation; D048429:Cell Size; D003521:Cyclopropanes / Q000737:chemistry / Q000494:pharmacology*; D003847:Deoxyglucose / Q000378:metabolism; D005227:Fatty Acids / Q000032:analysis; D005229:Fatty Acids, Monounsaturated / Q000737:chemistry / Q000494:pharmacology*; D005434:Flow Cytometry; D007328:Insulin / Q000494:pharmacology; D051379:Mice; D013230:Stearoyl-CoA Desaturase / Q000235:genetics / Q000378:metabolism*; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects / Q000201:enzymology
chemical_list: D003521:Cyclopropanes; D005227:Fatty Acids; D005229:Fatty Acids, Monounsaturated; D007328:Insulin; D003847:Deoxyglucose; D013230:Stearoyl-CoA Desaturase; C003345:sterculic acid