N-acetyl cysteine inhibits H2O2-mediated reduction in the mineralization of MC3T3-E1 cells by down-regulating Nrf2/HO-1 pathway

• DOI: 10.5483/bmbrep.2015.48.11.112
• Publication Date: 2016-05-09T07:14:08Z
• Authors: Daewoo Lee; Sung-Ho Kook; Hyeok Ji; Seung-Ah Lee; Ki-Choon Choi; Kyung-Yeol Lee; Jeong-Chae Lee
• Keywords: 0301 basic medicine; Osteoblasts; NF-E2-Related Factor 2; Osteocalcin; Down-Regulation; Cell Differentiation; Core Binding Factor Alpha 1 Subunit; 3T3 Cells; Hydrogen Peroxide; Acetylcysteine; Mice; Oxidative Stress; 03 medical and health sciences; Osteogenesis; Animals; Humans; Research-Article; Heme Oxygenase-1; Signal Transduction

There are controversial findings regarding the roles of nuclear factor (erythroid-derived 2)-like 2 (Nrf2)/heme oxygenase-1 (HO-1) pathway on bone metabolism under oxidative stress. We investigated how Nrf2/HO-1 pathway affects osteoblast differentiation of MC3T3-E1 cells in response to hydrogen peroxide (H2O2), N-acetyl cysteine (NAC), or both. Exposing the cells to H2O2 decreased the alkaline phosphatase activity, calcium accumulation, and expression of osteoblast markers, such as osteocalcin and runt-related transcription factor-2. In contrast, H2O2 treatment increased the expression of Nrf2 and HO-1 in the cells. Treatment with hemin, a chemical HO-1 inducer, mimicked the inhibitory effect of H2O2 on osteoblast differentiation by increasing the HO-1 expression and decreasing the osteogenic marker genes. Pretreatment with NAC restored all changes induced by H2O2 to near normal levels in the cells. Collectively, our findings suggest that H2O2-mediated activation of Nrf2/HO-1 pathway negatively regulates the osteoblast differentiation, which is inhibited by NAC.