Melengestrol acetate enhances adipogenic gene expression in cultured muscle-derived cells.

ABSTRACT

Melengestrol acetate (MGA) has been used in the United States for nearly 40 yr to enhance feedlot heifer performance, yet unequivocal studies have not been conducted to discover the mechanism of action. Our hypothesis was that MGA may induce various populations of muscle-derived cells (MDC) to the adipogenic pathway in both a bovine and murine cell culture model. To determine this, MDC were digested from the semimembranosus muscle tissue of six 14-mo-old crossbred steers. The addition of insulin, oleic acid, and ciglitizone (IOC) with cultured bovine MDC resulted in morphological differences compared with control cultures. Multilocular lipid droplets stained with Oil Red O were seen not only in single MDC, but also in fused myotubes. An increase (P < 0.05) in relative PPARgamma messenger RNA (mRNA) levels was measured in MDC incubated with IOC. However, myogenin mRNA levels in MDC incubated with IOC were repressed (P < 0.05) compared with nontreated MDC. Cultures of MDC treated with 10 microM insulin, 10 microM oleic acid, 10 microM ciglitizone, 10 nM estradiol-17beta (E2), and 10 nM MGA resulted in cultures with highly distributed lipid droplets not only in single cells, but also in the multinucleated myotubes. Relative C/EBPbeta and PPARgamma mRNA levels in total RNA isolated from MDC treated with MGA increased (P < 0.05) compared with control cultures. Estradiol treatment had no effect (P > 0.05) on these mRNA levels. The addition of both E2 and MGA to MDC increased (P < 0.05) C/EBPbeta mRNA levels and tended (P = 0.06) to increase the PPARgamma mRNA level. There was no difference (P > 0.10) in relative myogenin mRNA among the control, E(2), and MGA treatments. Relative C/EBPbeta, PPARgamma, and myogenin mRNA levels were investigated in murine C2C12, C3H 10T 1/2, and 3T3-L1 cells. Treatment of cultures with 10 nM MGA increased C/EBPbeta levels (P < 0.05) in C2C12 myoblasts and tended (P = 0.08) to increase C/EBPbeta levels in 3T3-L1 preadipocytes. These data indicate that populations of cells are present in postnatal skeletal muscle that, under the appropriate stimuli in a culture model, express adipogenic genes and accumulate lipids. In addition, the synthetic progestogen MGA appeared to upregulate the genes necessary for conversion to the adipogenic pathway.