Decrepitude and apoptosis of bone mesenchymal stem cells (BMSCs) induced by reactive oxygen species (ROS) lead to inhibited osteogenic differentiation, causing decreased bone density and osteoporosis. Quercetin, a bioactive component of Solanum muricatum extracts, promotes the osteogenic differentiation of BMSCs and ameliorates the symptoms of osteoporosis in vivo. However, the detailed mechanism underlying this process remains unclear. The study aims to reveal the regulatory mechanism of quercetin in BMSCs. Mouse BMSCs (mBMSCs) were isolated from the bone marrow and characterized by flow cytometry. QRT-PCR and western blot assays were performed to evaluate the expression levels of related genes and proteins. Alkaline phosphatase (ALP) staining and Oil Red O staining of lipids were used to estimate the osteogenesis and adipogenesis levels of mBMSCs, respectively. Quercetin treatment (2 and 5 μM) induced significant upregulation of antioxidant enzymes, SOD1 and SOD2, in mBMSCs. Quercetin promoted osteogenic differentiation and inhibited adipogenic differentiation of mBMSCs. Quercetin treatment enhanced the phosphorylation of AMPK protein and upregulated the expression of SIRT1, thus activating the AMPK/SIRT1 signaling pathway in mBMSCs. Quercetin promoted osteogenic differentiation and antioxidant responses of mBMSCs by activating the AMPK/SIRT1 signaling pathway.
Background:
Barbaloin, found in Aloe vera, exerts broad pharmacological activities including antioxidant, anti-inflammatory, and anti-cancer. This study aims to investigate the effects of barbaloin on the osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs).
Methods:
Osteogenic induction of hBMSCs was performed in the presence or absence of barbaloin. Then cell viability was determined by using CCK-8 assay. The characteristic of hBMSCs was identified by using Flow cytometry. Intracellular alkaline phosphatase (ALP) staining was performed to evaluate the ALP activity in hBMSCs. Alizarin Red S staining was performed to evaluate the matrix mineralization. The mRNA and protein levels of target genes were determined by using qRT-PCR and western blotting, respectively.
Results:
Treatment with barbaloin (10 and 20 μg/mL) significantly increased cell viability of hBMSCs after 72 hours. In addition, treatment with barbaloin increased the mRNA expression levels of ALP and its activities. Treatment with barbaloin also increased matrix mineralization and the mRNA and protein levels of late-differentiated osteoblast marker genes BMP2, RUNX2, and SP7 in hBMSCs. The underlying mechanisms revealed that barbaloin increased the protein expressions of Wnt/β-catenin pathway-related biomarkers.
Conclusion:
Barbaloin promotes osteogenic differentiation of hBMSCs by the regulation of the Wnt/β-catenin signaling pathway.
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