Effects of ursolic acid on proliferation and differentiation of bone marrow mesenchymal stem cells

doi:10.3969/j.issn.2095-4344.1729

Abstract

Abstract:

BACKGROUND: Ursolic acid can inhibit the proliferation and differentiation of osteoclasts through Wnt signaling pathway, to exert anti-inflammatory and osteoprotective effects in rats with collagen-induced arthritis. However, it is unclear about the effects of ursolic acid on proliferation and differentiation of rat bone marrow mesenchymal stem cells and expression of Wnt/β-catenin signaling pathway-related genes and proteins.
OBJECTIVE: To investigate the effects of ursolic acid on proliferation, differentiation and Wnt signaling pathway of bone marrow mesenchymal stem cells.
METHODS: Rat bone marrow mesenchymal stem cells were cultured in vitro by whole bone marrow adherence method, and then different concentrations of ursolic acid (1.0, 2.5, 5.0, 10.0, 20.0 μmol/L) acted on the cells. Cells with no treatment of ursolic acid were used as control group. MTT assay was used to detect the effect of ursolic acid on the proliferation of bone marrow mesenchymal stem cells. Differentiation of bone marrow mesenchymal stem cells was induced by osteogenic induction solution containing different concentrations of ursolic acid (1.0, 2.5, 5.0 μmol/L). Cells cultured in the osteogenic induction medium with no ursolic acid. Alkaline phosphatase activity and osteocalcin level were detected by ELISA at 7 and 14 days. RT-PCR was used to detect the mRNA levels of ALP, COL-I, OCN, OPN and RUNX2. The expressions of Wnt-3α, GSK-3β and β-catenin mRNAs and proteins related to Wnt signaling pathway were detected by RT-PCR and western blot, respectively.
RESULTS AND CONCLUSION: (1) Different concentrations of ursolic acid could promote the proliferation of bone marrow mesenchymal stem cells, and the proliferative rate was highest when the cells were treated with 5.0 μmol/L ursolic acid. (2) Compared with the control group, the alkaline phosphatase activity and osteocalcin level were significantly higher at 7 and 14 days after osteoinduction (P < 0.05), and the alkaline phosphatase activity and osteocalcin level increased with the increasing concentration of ursolic acid. (3) At 14 days after osteoinduction, ursolic acid promoted the expression of osteogenic genes ALP, COL-I, OCN, OPN, RUNX2 mRNAs and Wnt signaling pathway-related Wnt-3α, GSK-3β and β-catenin mRNAs and proteins and the expression levels increased with the increasing concentration of ursolic acid (P < 0.05). To conclude, ursolic acid can promote the differentiation of bone marrow mesenchymal stem cells into osteoblasts via Wnt/β-catenin signaling pathway.

Key words: ursolic acid, bone marrow mesenchymal stem cells, cell proliferation, osteogenic differentiation, alkaline phosphatase, osteocalcin, osteogenic gene, Wnt signaling pathway

CLC Number:

Liu Yaohui, Huang Zhenming, He Jingxuan, Huang Linghua, Li Jianping. Effects of ursolic acid on proliferation and differentiation of bone marrow mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(17): 2665-2671.

Figures/Tables 3

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