Abstract
BackgroundQuercetin and H19 can promote osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). However, whether quercetin regulates H19 expression to promote osteogenic differentiation of BMSCs is unclear.MethodsBMSC proliferation, matrix mineralization, and alkaline phosphatase (ALP) activity were assessed using the Cell Counting Kit-8, ALP assay kit, and alizarin red staining kit, respectively. Expression of H19, miR-625-5p, BMP-2, osteocalcin, and RUNX2 were measured by qRT-PCR; β-catenin protein level was measured by western blotting.ResultsQuercetin promoted BMSC proliferation, enhanced ALP activity, and upregulated the expression of BMP-2, osteocalcin, and RUNX2 mRNAs, suggesting that it promoted osteogenic differentiation of BMSCs. Moreover, quercetin increased H19 expression, while the effect of quercetin on BMSCs was reversed by silencing H19 expression. Additionally, miR-625-5p, interacted with H19, was downregulated during quercetin-induced BMSC osteogenic differentiation, which negatively correlated with H19 expression. Silencing miR-625-5p expression promoted BMSC proliferation and osteogenic differentiation, whereas miR-625-5p overexpression weakened the effect of quercetin on BMSCs. Finally, quercetin treatment or downregulation of miR-625-5p expression increased β-catenin protein level in BMSCs. Upregulation or downregulation of miR-625-5p or H19 expression, respectively, inhibited β-catenin protein level in quercetin treated-BMSCs.ConclusionH19 promotes, while miR-625-5p inhibits BMSC osteogenic differentiation. Quercetin activates the Wnt/β-catenin pathway and promotes BMSC osteogenic differentiation via the H19/miR-625-5p axis.
Highlights
Quercetin and H19 can promote osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs)
alkaline phosphatase (ALP) activity and the transcription of bone morphogenetic protein 2 (BMP-2), osteocalcin, and Runt-related transcription factor 2 (RUNX2) was significantly enhanced in the quercetin treatment and positive groups compared with those in the blank group (Fig. 2A–D)
These results suggest that quercetin treatment significantly increased cell proliferation and osteogenic differentiation
Summary
Quercetin and H19 can promote osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). Whether quercetin regulates H19 expression to promote osteogenic differentiation of BMSCs is unclear. Human bone marrow mesenchymal stem cells (BMSCs) can undergo self-renewal and osteogenic differentiation, and play a vital function for bone formation and remodeling [1]. Some pathophysiological conditions, such as aging, osteoporosis, and some bone defects can inhibit the osteogenic ability of BMSCs [2,3,4]. Quercetin promotes osteogenic differentiation of BMSCs via extracellular-signal-regulated protein kinases, adenosine 5’-monophosphate (AMP)-activated protein kinase/sirtuin 1, and estrogen receptor-mediated pathways [6,7,8]. Further understanding of the mechanisms underlying quercetin effects will have immense clinical implications
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