Effect of Angelica sinensis polysaccharide on the osteogenic differentiation of bone marrow mesenchymal stem cells of rats with high glucose levels

doi:10.7518/hxkq.2019.02.012

Abstract

Abstract:

Objective This study aims to evaluate the effect of Angelica sinensis polysaccharide (ASP) on the osteogenic differentiation of the bone marrow mesenchymal stem cells (BMSCs) of rats with high glucose levels. Methods Rat BMSCs were isolated and identified by osteogenic and adipo-genic differentiation. Then, the BMSCs were divided into three groups as follows: normal control group (5.5 mmol·L ^-1glucose), high glucose group (25.5 mmol·L ^-1glucose), and ASP+high glucose group (25.5 mmol·L ^-1 glucose +40 mg·L ^-1 ASP). The proliferation activities of the BMSCs were detected by CCK8. Alizarin red staining, and alkaline phosphatase activity were used in the examination of osteogenic activity. Quantitative real time-polymerase chain reaction was used to detect the expression levels of the osteogenic genes (Runx2, Osx, OCN, Col-Ⅰ) and the key factors of Wnt/β-catenin signal pathway (CyclinD1, β-catenin). In vivo, a type 2 diabetes rat model was established. The rats were divided into three groups, namely, the normal control group (normal rats), diabetes group (diabetic rats), diabetes+ASP group (diabetic rats, ASP feeding). Then, the tibia bone defect was established. The repair of bone defects in each group was observed through histological examination. Results The proliferation of BMSCs was higher in the high glucose group and ASP+high glucose group than in the normal control group (P<0.05). No significant difference was observed between the high glucose group and ASP+high glucose group (P>0.05). The number of calcium nodules of BMSCs; alkaline phosphatase activity; and the mRNA expression of Runx2, OCN, Osx, Col-Ⅰ, CyclinD1, β-catenin in the high glucose group were lower than those in the normal control and ASP+high glucose groups (P<0.05). No significant difference was observed between the normal control and ASP+high glucose groups (P>0.05). The bone mass was significantly lower in the bone defect of the diabetes group than in the bone defect of the normal control or diabetes+ASP group (P<0.05). No statistical difference was found between the normal control and diabetes+ASP groups (P>0.05). Conclusion ASP can promote the osteogenic differentiation of rat BMSCs under high glucose culture and induce bone regeneration in rats with type 2 diabetes. These features may be related to the activation of the Wnt/β-catenin signaling pathway.

Key words: Angelica sinensis polysaccharide, bone marrow mesenchymal stem cells, osteogenic differentiation, type 2 diabetes mellitus

CLC Number:

Q254

Feng Liao,Yao Liu,Hanghang Liu,Jian Hu,Shuang Zhao,Shimao Yang. Effect of Angelica sinensis polysaccharide on the osteogenic differentiation of bone marrow mesenchymal stem cells of rats with high glucose levels[J]. West China Journal of Stomatology, 2019, 37(2): 193-199.

Figures/Tables 7

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基因	引物序列（5’-3’）
Runx2	F：GCGGACGAGGCAAGAGTT
	R：TTGGTGCTGAGTTCAGGGAG
Osx	F：CTGTGAAACCTCAAGTCCTATGGA
	R：GCTCTGCAGTCAAGGGAGATG
OCN	F：CAGGTGCAAAGCCCAGCGACT
	R：AGGGGATCTGGGTAGGGGGCT
Col-Ⅰ	F：CCACCTGCCTCTGGCTTCT
	R：AGCTGTGGAGGAGGGTTTCA
β-catenin	F：TGGCAACCAAGAAAGCAAG
	R：CTGAACAAGAGTCCCAAGGAG
CyclinD1	F：CCCTCGGTGTCCTACTTCA
	R：GTTTGTTCTCCTCCGCCTCT
GAPDH	F：AGACCTTCAACACCCCAG
	R：CACGATTTCCCTCTCAGC