Effects of zeranol on protein turnover in L6 myotubes

Abstract

Protein synthesis and degradation were measured in cultures of L6 myotubes to determine the direct anabolic activity of zeranol on muscle. Zeranol, dexamethasone, insulin and zeranol-dexamethasone combination, at various concentrations from 10 −8 to 10 −6M, were added to cultures at either 18 hr prior to or at the beginning of a 6 hr synthesis or degradation measuring period. Protein synthesis was measured by determining the incorporation of radioactivity into trichloraoacetic acid precipitable cell protein following incubation with [ 3H] leucine. Protein synthesis was expressed as cpm incorporated in 6 hr per mg protein. Protein degradation was measured by a pulse-chase procedure using [ 3H] leucine. Protein degradation was expressed as the percent labeled protein degraded in 6 hr. Results from the study indicate that zeranol did not stimulate protein synthesis or inhibit proteolysis (P>.01). Stimulation of proteolysis observed with 10 −8M dexamethasone was 13% and 18% (P<.01) at the 6 hr and 24 hr incubation period, respectively. Dexamethasone-stimulated protein degradation was not altered appreciably by zeranol. In contrast, 10 −6M insulin significantly (P<.01) stimulated protein synthesis (16%) and inhibited protein degradation (15%). These results suggest that the anabolic action of zeranol does not occur by directly regulating muscle protein synthesis or degradation, or by altering the glucocorticoid-induced catabolic response in muscle.