Effect of p-ecdysterone on glucocorticoid-induced apoptosis and autophagy in osteoblasts

Abstract

The aim of the present study was to investigate the effect of glucocorticoids in osteoblasts and to examine the role of β-ecdysterone in the pathogenesis of glucocorticoid-induced osteoporosis. Osteoblasts were induced from bone marrow mesenchymal stem cells, which were isolated from C57BL/6 mice. Cell viability and apoptosis of osteoblasts were measured by Cell Counting Kit-8 and flow cytometry analysis, respectively. The expression of related genes and proteins was measured by reverse transcription quantitative polymerase chain reaction and western blot analysis respectively. Dose-dependent decreases in the cell proliferation and differentiation were observed in dexamethasone (Dex)-treated osteoblasts, evidenced by downregulation in the activity of alkaline phosphatasedecreased expression levels of Runt-related transcription factor 2 and osteocalcin, and upregulated expression of RANK ligand. Dex also induced apoptosis and inhibited autophagy of osteoblasts, evidenced by upregulated B-cell lymphoma 2 (Bcl-2)-associated X protein/Bcl-2 ratio and the activation of mammalian target of rapamycin (mTOR), and decreased expression levels of Beclin-1, autophagy protein 5 and microtubule-associated protein 1 light chain 3 II. The effects on cell proliferation, apoptosis and autophagy induced by Dex were reversed by β-ecdysterone in a dose-dependent manner. Therefore, β-ecdysterone may be a promising candidate drug for the treatment of osteoporosis through inducing osteoblast autophagic activity by inactivating mTOR.