Title:Hydroxysafflor Yellow A-Induced Osteoblast Differentiation and
Proliferation of BM-MSCs by Up-Regulating Nuclear Vitamin D
Receptor
Volume: 23
Issue: 5
Author(s): Jiewen Pan, Youwei Bao, Shuqing Pan, Danyan Zhuang, Yanan Xu, Xiaoli Pan and Haibo Li*
Affiliation:
- Key Laboratory of Comprehensive Prevention and Treatment of Congenital Anomalies, Women's and
Children's Hospital of Ningbo, Ningbo, China
Keywords:
HSYA, BM-MSCs, VDR, calcium deposition, proliferation, osteoblast.
Abstract:
Background: Vitamin D receptor (VDR) is critical for mineral and bone
homeostasis since it plays an essential role in the osteoblast differentiation of bone
marrow mesenchymal stem cells (BM-MSCs). Hydroxysafflor yellow A (HSYA) has the
potential to promote bone mineralization and inhibit bone resorption, while its detailed
mechanism needs to be elaborated.
Objective: This study intends to explore the action of HSYA on the proliferation and
differentiation of BM-MSC and the underlying mechanism.
Methods: Different concentrations of HSYA to BM-MSC and CCK-8, and EdU were
used to detect cell viability and proliferation. The alkaline phosphatase (ALP) was used
to observe the differentiation ability of BM-MSC osteoblasts. The calcium uptake and
mineralization of osteoblast-like cells were observed by alizarin red staining. The level of
calcium ion uptake in cells was detected by flow cytometry. AutoDock was performed for
molecular docking of HSYA to VDR protein. Immunofluorescence and western blotting
were performed to detect the expression of VDR expression levels. Finally, the effect of
VDR was verified by a VDR inhibitor.
Results: After treatment with HSYA, the proliferation and calcium uptake of BM-MSC
were increased. The level of ALP increased significantly and reached its peak on the
12th day. HSYA promoted calcium uptake and calcium deposition, and mineralization of
osteoblasts. The western blotting and immunofluorescence showed that HSYA
increased the expression of VDR in the osteoblast-like cell's nucleus and upregulated
Osteocalcin, S100 calcium-binding protein G, and CYP24A1. In addition, HYSA
treatment increased the expression of osteopontin and the synthesis of osteogenic
proteins, such as Type 1 collagen. After the addition of the VDR inhibitor, the effect of
HSYA was weakened.
Conclusion: HSYA could significantly promote the activity and proliferation of
osteoblasts and increase the expression level of VDR in osteoblasts. HSYA may also
improve calcium absorption by osteoblasts by regulating the synthesis of calciumbinding
protein and vitamin D metabolic pathway-related proteins.
