Tauroursodeoxycholate

A series of bile acid standards, containing cholate (CA), chenodeoxycholate (CDCA), ursodeoxycholate (UDCA), deoxycholate (DCA), lithocholate (LCA), glycocholate (GCA), glycochenodeoxycholic acid (GCDCA), glycodeoxycholate (GDCA), taurocholate (TCA), taurodeoxycholate (TDCA), tauroursodeoxycholate (TUDCA), taurohyodexoycholate (THDCA), taurochenodeoxycholate (TCDCA), taurolithocholate (TLCA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Tauro-β-muricholate (TβMCA) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). As internal standard (IS), deoxycholic acid-2,2,4,4-d4 (DCA-d4) were obtained from CDN isotopes (Pointe-Claire, Quebec, Canada). All organic reagents for mass spectrometric analysis were HPLC grade purchased from Sigma-Aldrich (St. Louis, MO, USA). And spexin used for drug treatment was purchased from Phoenix Pharmaceuticals (Belmont, CA, USA). M871 was purchased from R&D Systems (Minneapolis, MN, USA). SNAP37889 was purchased from Key Organics Ltd (Camelford, Cornwall, UK).

The generation of immortalized preadipocytes from the stromal vascular fraction of inguinal white adipose tissue has been reported previously^{31 (link)}. Cells were grown to confluence in culture medium comprising Dulbecco’s modified Eagle medium (4.5 g/l glucose, GE Healthcare Bio-Sciences), 20% fetal bovine serum (Life Technologies), 20 nM insulin and 1 nM T3. Adipocyte differentiation was induced by complementing the medium with 250 µM indomethacin, 500 µM isobutylmethylxanthine and 2 µg/ml dexamethasone for 24 hours after confluence. During six days of differentiation, the medium was supplemented or not (control) with one of the following compounds (each at 20 µM): rosiglitazone (Biomol), cholate, glycodeoxycholate, ursodeoxycholate, taurodeoxycholate, chenodeoxycholate, taurocholate, deoxycholate, tauroursodeoxycholate, taurochenodeoxycholate, glyocholate (all Sigma-Aldrich) or tauro-beta-muricholate (Santa Cruz). Primary brown adipocytes were isolated, cultured and differentiated from the stromal vascular fraction of interscapular BAT of male 129S6/SvEvTac mice as described previously^{13 (link)}.