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TSG Targeting KDM5A affects osteogenic differentiation of bone mesenchymal stem cells induced by bone morphogenetic protein 2

By Min Wei, Yi Jiang, Yuanqing Huang

Posted 25 Jul 2020
bioRxiv DOI: 10.1101/2020.07.25.221424

To investigate the effect of 2, 3, 5, 4'-tetrahydroxystilbene-2-O-β-D-glucoside (TSG) on osteogenic differentiation of bone marrow mesenchymal stem cells (BMSC) and its molecular mechanism. The effects of TSG on alkaline phosphatase positive cloning and mineralized nodule formation were also detected. Total mRNA and protein were extracted and 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. TSG significantly increased the activity of ALP and 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. The mechanism study showed that the effect of TSG is realized by inhibiting KDM5A. KDM5A signaling may be involved in the regulation of osteogenic differentiation of rBMSC. TSG can promote osteogenic differentiation and maturation of rBMSC at 0.1-50 μmol/L. The mechanism of action was realized by inhibiting the expression of KDM5A.

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