Inhibition of Stearoyl-CoA Desaturase Enzyme Alters Oleate Turnover In NIH3T3 Preadipocytes.

Stearoyl-CoA desaturase (SCD) is a key enzyme in the conversion of stearate to oleate (C18:0 to C18:1), and palmitate to palmitoleate (C16:0 to C16:1). There are two mouse SCD genes (SCD1 and SCD2). While SCD 1 is the isoform of SCD highly expressed in the liver and adipose tissue, and its expression is induced during adipocyte differentiation with dexamethasone and insulin, SCD2 is constitutively expressed in preadipocytes. The objective of the study is to evaluate the effect of SCD inhibition on fatty acid composition and turnover in NIH3T3 preadipocytes. NIH3T3 preadipocytes were grown in culture for 72 hours with [U-[sup 13]C] stearate and [1, 2 [sup 13]C]-acetate with 1) DMSO or 2) 0.8 µM of SCD inhibitor 06811M1005 (generously provided by CompleGen, Inc.). Analysis of fatty acid mass isotopomers was performed by gas chromatography/mass spectroscopy (GCMS). Fatty acid composition and the desaturation indices were calculated from the gas chromatogram. In cells cultured with SCD inhibitor 06811M1005 in [U-[sup 13]C]-stearate and [1,2 [sup 13]C]-acetate, the desaturation index for C18:1 n-9 was significantly reduced by ∼30% (p<0.03). There was a reduction in the conversion of U-stearate to U-oleate from 64% to 38% (p<0.04) when SCD was inhibited. While 13C isotopomer enrichment of palmitate and stearate reached the same plateau levels in treated and untreated cells, 13C isotopomer enrichment of oleate in the treated cells was significantly lower than control suggesting reduced oleate turnover during the study period. We conclude: NIH3T3 preadipocytes differentiate either in the presence of dexamethasone and insulin or PPAR υ agonist into adipocytes with phenotypes having different expression of SCD1. SCD inhibitor 06811M 1005 probably acts equally on SCD1 and SCD2 in the preadipocytes resulting in a change in desaturation index. Contrary to the expectation of reduced de novo lipogenesis, SCD inhibition primarily reduced on the conversion of stearate to oleate and the turnover of oleate without affecting the uptake and de novo synthesis of palmitate or stearate. [ABSTRACT FROM AUTHOR]