Quercetin stimulates osteogenic differentiation of bone marrow stromal cells through miRNA-206/connexin 43 pathway. 2020

Qihao Zhang, and Bo Chang, and Guizhou Zheng, and Shixin Du, and Xuedong Li
Department of Orthopedics, The Third Affiliated Hospital (The Affiliated Luohu Hospital) of Shenzhen University Shenzhen 518000, Guangdong, P. R. China.

The quantity and function of osteoblasts require continuous osteogenic differentiation of bone marrow mesenchymal stem cells. Recent evidence suggests that microRNAs (miRNAs) act as important post-transcriptional regulators in a wide range of biological processes, including osteoblastic differentiation. Quercetin has also been found to prevent bone loss. In this study, we investigated the osteogenesis of quercetin and miR-206 on bone marrow mesenchymal stem cells (BMSCs) and their relationship. We observed quercetin enhanced BMSCs proliferation with a dose-dependent manner in Cell Counting Kit-8 (CCK-8). Alizarin red S staining, alkaline phosphatase (ALP) quantification assay, miR-206 and mRNA levels of osteogenesis marker genes by quantitative real-time PCR (qPCR) were used to analyze osteogenic potential. We observed quercetin significantly elevated bone mineralization and the mRNA expression levels of osteoblast-specific genes including Runt-related transcription factor 2 (Runx2), Osterix (OSX), osteocalcin (OCN), and osteopontin (OPN). Correspondly, Cx43 expression were increased, while miR-206 expression were decreased. In the presence of agomir of miR-206, effects of quercetin on mineralization, alkaline phosphatase activity and osteoblast-specific genes expression were suppressed. Most of all, Cx43 protein level was also blocked while overexpression of miR-206 against quercetin effects. Taken together, these data indicated that quercetin promotes BMSCs proliferation and osteogenic differentiation. The osteogenic effect of quercetin is partly modulated through miR-206/Cx43 pathway.

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