Enhancing effect of troglitazone on porcine adipocyte differentiation in primary culture: a comparison with dexamethasone.
OBJECTIVE: This study compares the effects of the thiazolidinedione, troglitazone (TGZ), dexamethasone (DEX), and DEX plus TGZ on preadipocyte differentiation and the expression of CCAAT/enhancer-binding protein-alpha (C/ EBPalpha) and peroxisome proliferator-activated receptor-gamma (PPARgamma). RESEARCH METHODS AND PROCEDURES: Adipose tissue was obtained from postnatal pigs to isolate stromal-vascular cells. First, we applied 1, 5, or 10 microM TGZ and 10% fetal bovine serum for 3 days and counted the number of recruited preadipocytes. Next, we used either 10 microM TGZ, 80 nM DEX, or DEX plus TGZ with 10% fetal bovine serum for 3 days and then switched to serum-free medium with insulin for 6 days. On day 3 of culture, we counted preadipocytes, and on days 3 and 6 of culture, we performed immunostaining and Western blot analysis to determine the expression of C/EBPalpha and PPARgamma proteins. On day 9 of culture, we stained for lipids with oil red-O and measured the activity of glycerol phosphate dehydrogenase. RESULTS: DEX and TGZ equally enhanced recruitment of preadipocytes and late differentiation, but these effects were not additive with DEX plus TGZ treatment. However, TGZ and DEX had a differential effect on morphogenesis; DEX-treated adipocytes were larger and organized in loose clusters, whereas TGZ-treated cells were smaller and formed compact clusters. Both agents increased C/EBPalpha expression but in a temporally distinct manner. DEX was a better inducer than TGZ, and its effect was early and temporary. However, treatment with either TGZ or DEX did not change PPARgamma protein expression as evaluated by a Western blotting, but immunocytochemistry showed a tendency for increased numbers of PPARgamma positive cells. DISCUSSION: TGZ and DEX equally enhance early and late adipocyte differentiation, possibly by using some common pathways for preadipocyte recruitment. The differential effect on morphogenesis implies a potential differential effect on the expression of extracellular matrix components. C/EBPalpha may be the critical transcription factor involved in TGZ- and DEX-induced adipogenesis.[1]References
- Enhancing effect of troglitazone on porcine adipocyte differentiation in primary culture: a comparison with dexamethasone. Tchoukalova, Y.D., Hausman, D.B., Dean, R.G., Hausman, G.J. Obes. Res. (2000) [Pubmed]
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