Abstract
Objectives To investigate the effect of 2,3,5,4′-tetrahydroxystilbene-2-O-β-D-glucoside (TSG) on the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) and its molecular mechanism. Methods After TSG treatment of rBMSCs, alkaline phosphatase (ALP) activity was compared between the drug treatment group and the control group. The effects of TSG on alkaline phosphatase positive cloning and mineralized nodule formation were also detected. Total mRNA and protein were extracted, and the effects of TSG on the expression levels of osteopontin (OPN), osteocalcin (OCN), Runt-related transcription factor 2 (Runx2), Osterix, and Col1a1 were detected by real-time fluorescence quantitative PCR. Western blotting was used to detect the inhibitory effect of TSG on KDM5A. BMSCs were transfected with small interfering RNA (siRNA) targeting KDM5A (si-KMD5A) and pcDNA3.1 KMD5A. Results TSG significantly increased the activity of ALP, the number of alkaline phosphatase clones, and calcified nodule formation. The OPN, OCN, Runx2, and Osterix expression levels were significantly increased among the osteoblasts after TSG treatment. A mechanistic study showed that the effect of TSG is realized by inhibiting KDM5A. Conclusions KDM5A signaling may be involved in the regulation of osteogenic differentiation of rBMSCs. TSG can promote osteogenic differentiation and maturation of rBMSCs at 0.1–50 μmol/L. The mechanism of action was realized by inhibiting the expression of KDM5A.
Highlights
Osteoporosis (OP) is a metabolic bone disease characterized by reduced bone mass, damaged bone mass, and reduced bone strength, resulting in increased bone fragility and proneness to fracture
We evaluated the effect of TSG on alkaline phosphatase in rBMSCs
Further experimental results showed that different concentrations of TSG increased alkaline phosphatase (ALP) activity (Figure 1(d)) and presented a concentrationdependent effect. e influence of TSG on the formation of calcified nodules showed that both the control group and the TSG group formed calcified nodules after osteogenic induction of rBMSCs
Summary
Osteoporosis (OP) is a metabolic bone disease characterized by reduced bone mass, damaged bone mass, and reduced bone strength, resulting in increased bone fragility and proneness to fracture. Bone marrow mesenchymal stem cells (BMSCs) are derived from bone marrow and have the function of selfrenewal and multidifferentiation [1]. E osteogenic differentiation ability of BMSCs decreases with age [2, 3]. The proliferation and self-renewal capacity of MSCs are reduced, the number of apoptotic cells is increased, osteogenic differentiation is weakened, lipogenic differentiation is enhanced, and the regulation of multidirectional differentiation is disordered [4,5,6]. TSG is an active component of Polygonum multiflorum and has a wide range of biological activities [7,8,9].
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