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| Epimedium-derived flavonoids modulate the balance between osteogenic differentiation and adipogenic differentiation in bone marrow stromal cells of ovariectomized rats via Wnt/β-catenin signal pathway activation |
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| Author Name | Affiliation | E-mail | | Ying-xing Xu | Research Institution of Orthopaedics and Traumatology, Zhejiang Traditional Chinese Medical University, Hangzhou, 310053, China
Department of Orthopaedics and Traumatology, Chinese Medicine Hospital, Yuxi, 653100, Yunnan Province, China | xuyingxing5192@126.com | | Cheng-liang Wu | Research Institution of Orthopaedics and Traumatology, Zhejiang Traditional Chinese Medical University, Hangzhou, 310053, China | | | Yan Wu | The First Clinical Medical College, Zhejiang University of Traditional Chinese Medicine, Hangzhou, 310053, China | | | Pei-jian Tong | Department of Orthopaedics and Traumatology, Chinese Medicine Hospital, Hangzhou, 310053, China | | | Hong-ting Jin | Research Institution of Orthopaedics and Traumatology, Zhejiang Traditional Chinese Medical University, Hangzhou, 310053, China | | | Nan-ze Yu | Research Institution of Orthopaedics and Traumatology, Zhejiang Traditional Chinese Medical University, Hangzhou, 310053, China | | | Lu-wei Xiao | Research Institution of Orthopaedics and Traumatology, Zhejiang Traditional Chinese Medical University, Hangzhou, 310053, China | |
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| Abstract: |
Objective To observe the function of wnt/??-catenin signal pathway on the process that epimedium-derived flavonoids (EFs) regulate the balance between osteogenic differentiation and adipogenic differentiation in bone marrow stromal cells of ovariectomized rats, and to provide an experimental evidence for the mechanism of EFs on treating postmenopausal osteoporosis. Methods Bone marrow stromal cells from ovariectomized rats were separated and cultivated in the condition of osteoinductive medium or liquid medium for 15 days. Low- (1 ??g/mL), medium- (10 ??g/mL) and high- (100 ??g/mL) dose EFs were administrated correspondingly. Alkaline phosphatase (ALP) staining, ALP activity determination, oil red O staining and realtime polymerese chain reaction (RT-PCR) were used to determine the effect of EFs on osteogenic differentiation and adipogenic differentiation in bone marrow stromal cells of ovariectomized rats. Moreover, in order to explore the mechanism of EFs on osteogenic differentiation and adipogenic differentiation in bone marrow stromal cells of ovariectomized rats, Dickkopf-related protein 1 (DKK1) was used in the medium group. Enzymelinked immunosorbent assay (ELISA) and RT-PCR were used to determine mRNA levels of ??-catenin, low density lipoprotein receptor-related protein 5 (LRP5) and T cell factor (TCF) protein, known as wnt/??-catenin signal pathway related factors. Results EFs increased mRNA expression levels of ALP and early osteoblast differentiation factors, such as runt-related transcription factor 2 (Runx2), osteocalcin and collagen I, and decreased mRNA expression levels of fat generation factors, such as peroxisome proliferator activated receptor gamma 2 (PPAR??-2) and CCAAT enhancer-binding protein-?? (C/EBP??) in a dose-dependent manner. While osteoblast differentiation factors were down-regulated, fat generation factors were up-regulated when DKK1 was applied. Also EFs up-regulated mRNA expression levels of ??-catenin, LRP5 and TCF protein which could be blocked by DKK1. Conclusion EFs regulate the balance between osteogenic differentiation and adipogenic differentiation in bone marrow stromal cells of ovariectomized rats by activating wnt/??-catenin signal pathway, which may be an important molecular mechanism of EFs on treating postmenopausal osteoporosis. |
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