Induction of apoptosis by all-trans-retinoic acid and C2-ceramide treatment in rat stromal-vascular cultures.

Apoptosis of preadipocytes and adipocytes contributes to the balance of adipose tissue mass by reducing adipocyte number. To address this phenomenon, we treated cultured rat S-V cells with all-trans-retinoic acid (RA) (10 microM) or C2-ceramide (50 microM) during adipogenesis. Gel electrophoresis of DNA from treated cells cultured in serum-free medium showed that 10 microM RA or 50 microM ceramide induced a distinct laddering pattern of DNA fragments. Cellular caspase 3 activity, another marker of apoptosis, was increased by RA (10 microM) (P < 0.05), but not by 50 microm C2-ceramide. RT-PCR results showed that RA (10 microM) decreased the expression of Bcl-2 mRNA. These results suggest that fat cell loss by apoptosis can be regulated, in part, by RA (10 microM) which increases caspase 3 activity and decreases Bcl-2 expression in rat S-V cells. C2-ceramide apparently works through a different cellular mechanism to induce apoptosis. Copyright 2000 Academic Press.