Hongping Yu1,2, Kaiyang Wang1, Pei Liu1, Pengbo Luo1, Daoyu Zhu1, Junhui Yin3, Qianhao Yang1, Yigang Huang1, Junjie Gao1, Zisheng Ai4, Yixuan Chen1, Youshui Gao1. 1. Department of Orthopedic Surgery, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China. 2. The First Affiliated Hospital of Xiamen University, Xiamen, China. 3. Institute of Microsurgery on Extremities, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China. 4. Department of Medical Statistics, Tongji University School of Medicine, Shanghai, China.
Abstract
OBJECTIVES: Alcohol consumption is one of the leading factors contributing to premature osteopenia. MicroRNA (miRNA) coordinates a cascade of anabolic and catabolic processes in bone homeostasis and dynamic vascularization. The aim was to investigate the protective role of miR-4286 in alcohol-induced bone loss and its mechanism. MATERIALS AND METHODS: The effect of miR-4286 and alcohol on bone mesenchymal stem cells (BMSCs) and human umbilical vein endothelial cells (HUVECs) was explored via multiple in vitro assays, including cell proliferation, QPCR, Western blot, osteogenesis, angiogenesis etc miR-4286 directly regulated HDAC3 was investigated by luciferase reporter assay, and the function of HDAC3 was also explored in vitro. Moreover, alcohol-induced bone loss in mice was established to reveal the preventive effect of miR-4286 by radiographical and histopathological assays. RESULTS: In vitro, ethanol dramatically inhibited the proliferation and osteogenesis of BMSCs, and substantially impaired the proliferation and vasculogenesis of HUVECs. However, a forced overexpression of miR-4286 within BMSCs and HUVECs could largely abolish inhibitory effects by alcohol. Furthermore, alcohol-induced inhibition on osteogenic and vasculogenic functions was mediated by histone deacetylase 3 (HDAC3), and dual-luciferase reporter assay showed that HDAC3 was the direct binding target of miR-4286. In vivo, micro-CT scanning and histology assessment revealed that miR-4286 could prevent alcohol-induced bone loss. CONCLUSIONS: We firstly demonstrated that miR-4286 might function via intimate osteogenesis-angiogenesis pathway to alleviate alcohol-induced osteopenia via targeting HDAC3.
OBJECTIVES:Alcohol consumption is one of the leading factors contributing to premature osteopenia. MicroRNA (miRNA) coordinates a cascade of anabolic and catabolic processes in bone homeostasis and dynamic vascularization. The aim was to investigate the protective role of miR-4286 in alcohol-induced bone loss and its mechanism. MATERIALS AND METHODS: The effect of miR-4286 and alcohol on bone mesenchymal stem cells (BMSCs) and human umbilical vein endothelial cells (HUVECs) was explored via multiple in vitro assays, including cell proliferation, QPCR, Western blot, osteogenesis, angiogenesis etc miR-4286 directly regulated HDAC3 was investigated by luciferase reporter assay, and the function of HDAC3 was also explored in vitro. Moreover, alcohol-induced bone loss in mice was established to reveal the preventive effect of miR-4286 by radiographical and histopathological assays. RESULTS: In vitro, ethanol dramatically inhibited the proliferation and osteogenesis of BMSCs, and substantially impaired the proliferation and vasculogenesis of HUVECs. However, a forced overexpression of miR-4286 within BMSCs and HUVECs could largely abolish inhibitory effects by alcohol. Furthermore, alcohol-induced inhibition on osteogenic and vasculogenic functions was mediated by histone deacetylase 3 (HDAC3), and dual-luciferase reporter assay showed that HDAC3 was the direct binding target of miR-4286. In vivo, micro-CT scanning and histology assessment revealed that miR-4286 could prevent alcohol-induced bone loss. CONCLUSIONS: We firstly demonstrated that miR-4286 might function via intimate osteogenesis-angiogenesis pathway to alleviate alcohol-induced osteopenia via targeting HDAC3.