Polygonatum sibiricum polysaccharide regulates osteoblastic differentiation of mouse bone marrow-derived mesenchymal stem cells

doi:10.3969/j.issn.2095-4344.2016.15.001

Abstract

Abstract:

BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) have the ability of multi-directional differentiation. Polygonatum sibiricum polysaccharide can promote osteogenetic differentiation of mouse BMSCs by activating Wnt/β-catenin signaling pathway, which is expected to become a new drug for the treatment of osteoporosis.

OBJECTIVE: To investigate the effects of Polygonatum sibiricum polysaccharide on Wnt/β-catenin signaling pathway in the osteogenic differentiation of mouse BMSCs.

METHODS: The mouse BMSCs were cultured and induced in osteoblast medium containing final concentrations (5, 10, 25, 50mg/L) of Polygonatum sibiricum polysaccharide. The mouse BMSCs treated without Polygonatum sibiricum polysaccharide was set as the negative control group. The morphological changes of cells were observed under an inverted microscope. Alkaline phosphatase (ALP) activity assay was performed by PNPP method. The mineralization nodules were observed and stained with alizarin red S and the number and area fraction were recorded under an inverted microscope. The mRNA expressions of osteogenesis-related genes ALP, Runx2, and osteocalcin were evaluated by quantitative real-time PCR (qRT-PCR). qRT-PCR and western blot were used to determine the expression level of β-catenin. The downstream β-catenin/TCF transcriptional activity was evaluated with the Dual-Luciferase Reporter Assay System.
RESULTS AND CONCLUSION: Compared with the control group, polygonatum sibiricum polysaccharide significantly enhanced the alkaline phosphatase activity, the mineralization ability of cells, and the mRNA expression of ALP, Runx2 and osteocalcin in the differentiated BMSCs in a dose dependent manner (P < 0.05). After induction, the mRNA expression of β-catenin was the highest on the 3^rd day. Polygonatum sibiricum polysaccharide significantly increased the expression of β-catenin (P < 0.05) in the process of promoting the differentiation of BMSCs into osteoblasts, and also promoted the high-level expression of luciferase reporter gene (TOPFlash) which contains wild type TCF binding sites (P < 0.05). These results demonstrate that Polygonatum sibiricum polysaccharide can promote the osteoblast differentiation of mouse BMSCs by activating the Wnt/β-catenin signaling pathway.
中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Polygonatum, Mesenchymal Stem Cells, Cell Differentiation, beta Catenin, TCF Transcription Factors, Tissue Engineering

Nong Meng-ni, Zeng Gao-feng, Zong Shao-hui, Du Li, Li Ke-ke, Peng Xiao-ming, Yan Fang-na. Polygonatum sibiricum polysaccharide regulates osteoblastic differentiation of mouse bone marrow-derived mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(15): 2133-2139.

Figures/Tables 4

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