Myostatin inhibits insulin-like growth factor 1-dependent citrate secretion and osteogenesis via nicotinamide adenine dinucleotide phosphate oxidase-4 in a mouse mesenchymal stem cell line.

Citrate is an indispensable component of bone. Reduced levels of citrate in bone and serum are reported in the elderly and in osteoporosis patients. Myostatin (Mstn) is implicated in skeletal homeostasis, but its effects on osteogenesis remain incompletely understood. Nox4 has critical roles in bone homeostasis. TGF-ß/Mstn-associated Smad2/3 signaling has been linked to Nox4 expression. Insulin-like growth factor (IGF-1) has been shown to counteract many regulatory effects of Mstn. However, the crosstalk among Mstn, IGF-1, and Nox4 is not well understood; the interactive effects of those factors on citrate secretion, osteogenic differentiation, and bone remodeling remain unclear. In this study, we demonstrated that osteogenic differentiation induced an IGF-1-dependent upregulation of citrate secretion that was suppressed by Mstn. Inhibition of Nox4 prevented Mstn-induced reduction of citrate secretion. In addition, Mstn reduced bone nodule formation; these changes were prevented by Nox4 inhibition. Moreover, Mstn increased the ratio of RANKL to OPG mRNAs to favor osteoclast activation. These results indicate that Mstn negatively regulates osteogenesis by increasing levels of Nox4, which reduced IGF-1 expression, citrate secretion, and bone mineralization while also altering the RANKL to OPG ratio. These findings provide new and highly relevant insights into the osseous effects of myostatin.

AdipoRon prevents myostatin-induced upregulation of fatty acid synthesis and downregulation of insulin activity in a mouse hepatocyte line.

Liver diseases such as non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH) are characterized by excess hepatic accumulation of lipid droplets and triglycerides which are associated with defective insulin action. Myostatin (Mstn) and adiponectin, secreted by muscle cells and adipocytes, respectively, play important roles in regulating insulin signaling and energy metabolism. The mechanisms underlying the actions of Mstn and adiponectin remain largely unknown. Moreover, the interactions between Mstn and adiponectin in regulating gene expression critical for fatty acid metabolism and insulin action in hepatocytes have not been investigated. The effects of Mstn and AdipoRon, a synthetic adiponectin receptor agonist that is orally active, alone or in combination, on hepatic gene expression and function was investigated. While Mstn increased fatty acid (FA) accumulation and desensitized cellular responses to insulin, AdipoRon protected against Mstn-induced defects in hepatic gene expression and function. In addition, these effects of Mstn were associated with reduced AMPK and PPARα activities which were reversed by AdipoRon. Finally, AdipoRon was able to prevent Mstn-induced activation of the Smad2/3 pathway. These data suggest crosstalk between Mstn-induced Smad2/3 and adiponectin-induced AMPK/PPARα pathways, which may play important roles in the regulation of hepatic gene expression critical for FA metabolism and insulin signaling. In addition, the data suggest that AdipoRon, as an adiponectin receptor agonist, may serve a therapeutic role to reduce the hepatic contribution to the disorders of fat metabolism and insulin action.