An artificial NEFA mixture based on the ratio and concentrations of FAs appearing in serum from obese rats reciprocally regulates β-catenin and PPARγ expression in ST2 cells.
(A), GC-MS analysis of NEFA. To measure NEFA composition, 100 µl of serum from LFD, MFD or HFD rats was characterized and quantified by a Shimazu QP-2010 GC-MS system after TLC separation. (B), Mean concentrations of the 5 major FAs in NEFA are presented. (C), ST2 cells were treated with a NEFA mixture based on individual FA concentrations appearing in serum in LFD, MFD and HFD rats for 24 and 48 h. Real-time RT-PCR of β-catenin and PPARγ mRNA expressions. (D) Western blot analysis of β-catenin and PPARγ (n = 3). LFD, low fat diet (control pelleted AIN-93G 14% fat diet); MFD, medium fat TEN diet (25% fat diet); HFD, high fat TEN diet (45% fat diet). Data bars are expressed as mean ± SEM (n = 3/treatment). *, P<0.05, versus control (vehicle treatment) as determined by t-test.