Valproic acid promotes osteogenic differentiation of rat bone marrow mesenchymal stem cells

doi:10.12307/2023.365

Abstract

Abstract: BACKGROUND: Valproic acid, as a histone deacetylase inhibitor, has the ability to promote osteogenic differentiation of cells, but the mechanism is still unclear.
OBJECTIVE: To investigate the effect and mechanism of valproic acid on osteogenic differentiation of rat bone marrow mesenchymal stem cells.
METHODS: Bone marrow mesenchymal stem cells were isolated from Sprague-Dawley rats and identified by flow cytometry. CCK-8 assay was used to detect the effects of different concentrations of valproic acid on the proliferation of rat bone marrow mesenchymal stem cells, and determine the used concentration of valproic acid. After adding valproic acid, rat bone marrow mesenchymal stem cells were induced to osteogenic differentiation. Alkaline phosphatase staining was performed on day 7 of osteogenic induction culture, and alizarin red staining was performed on day 21 of osteogenic induction culture. Quantitative real-time polymerase chain reaction was used to detect the expression of osteogenesis-related genes and EZH2 on days 3 and 7 of osteogenic induction culture. The protein expression levels of Runt-related transcription factor 2, EZH2 and H3K27me3 were detected by western blot assay on day 5 of osteogenic induction culture.
RESULTS AND CONCLUSION: (1) CCK-8 cell proliferation assay showed that bone marrow mesenchymal stem cell proliferation was significantly inhibited by increasing the concentration of valproic acid. (2) The results of alkaline phosphatase staining and alizarin red staining showed that valproic acid could increase the expression of alkaline phosphatase and enhance the ability of bone mineralization. (3) qRT-PCR results showed that valproic acid could increase the expression of osteogenesis-related genes and decrease the expression of EZH2 gene. (4) Western blot assay results showed that valproic acid could increase the expression of Runt-related transcription factor 2 protein and decrease the expression of EZH2 and H3K27me3 protein. (5) It is concluded that valproic acid can promote osteogenic differentiation of rat bone marrow mesenchymal stem cells by down-regulating EZH2 expression.

Key words: valproic acid, EZH2, H3K27me3, bone marrow mesenchymal stem cell, osteogenic differentiation

CLC Number:

Ling Xuwei, Sun Jie, Liu Chang, Wang Yi, Shi Qin, Yang Huilin. Valproic acid promotes osteogenic differentiation of rat bone marrow mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(15): 2304-2310.

Figures/Tables 6

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