Chinese Journal of Tissue Engineering Research ›› 2023, Vol. 27 ›› Issue (11): 1722-1727.doi: 10.12307/2022.1021

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Mechanism by which naringin regulates osteogenic differentiation in osteoblasts

Wu Jingjing1, Lin Haixiong2,3, Sun Weipeng1, Li Zige1, Jiang Ziwei4   

  1. 1The First Clinical Medical College of Guangzhou University of Chinese Medicine, Guangzhou 510405; 2Ningxia Chinese Medicine Research Center, Yinchuan 750000; 3Institute of Tissue Engineering and Regenerative Medicine, The Chinese University of Hong Kong, Sha Tin 999077, Hong Kong Special Administrative Region, China; 4The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510405
  • Received:2021-11-08 Accepted:2022-02-24 Online:2023-04-18 Published:2022-09-24
  • Contact: Jiang Ziwei, MD, Professor, Associate chief physician, The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China
  • About author:Wu Jingjing, Master candidate, The First Clinical Medical College of Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China
  • Supported by:
    the National Natural Science Foundation of China, No. 81774337 (to JZW)

Abstract: BACKGROUND: Studies have found that naringin affects bone metabolism directly through Wnt, transforming growth factor β, MAPK signaling pathway, and osteoclast signaling pathway, as well as indirectly through P13K-Akt and vascular endothelial growth factor signaling pathway. Naringin can promote bone repair and retard osteoporosis. 
OBJECTIVE: To investigate the effect of naringin on the proliferation and osteogenic differentiation of osteoblasts by targeting the activation of connexin 43/ERK1 signaling pathway through down-regulation of miR-206.
METHODS: Rat osteoblasts (ROB cells) were treated and cultured with different concentrations of naringin (1, 10, 100 mg/L and 1, 10 g/L), and the cell proliferation was detected by cell counting kit-8 assay. Naringin at 100 mg/L and 1, 10 g/L were used for subsequent experiments. Osteoblasts with non-drug culture were used as controls. RT-qPCR was used to detect the relative expression levels of miR-206, connexin 43 and ERK1 mRNAs in ROB cells to verify their targeting relationship. Western blot was used to detect the protein expression levels of ERK1/2 and P-ERK in ROB cells. Alkaline phosphatase activity and calcification ability in ROB cells were observed by alkaline phosphatase staining and alizarin red staining, respectively.
RESULTS AND CONCLUSION: Naringin significantly promoted the proliferation activity of ROB cells (P < 0.05). The absorbance value and relative proliferation rate of ROB cells increased as the administration concentration of naringin increased in a certain range. When the mass concentration was 1 g/L, the cell proliferation activity was the strongest. Naringin significantly inhibited the expression of miR-206 mRNA in ROB cells and promoted the expression of connexin 43 mRNA and ERK1/2 and P-ERK protein (P < 0.05). Naringin could significantly promote the activity and expression of alkaline phosphatase and the level of cellular calcification in ROB cells (P < 0.05). Moreover, naringin at 1 g/L showed the best effect. To conclude, naringin can target and activate the connexin 43/ERK1 signaling pathway by down-regulating the expression of miR-206, thereby promoting the proliferation and osteogenic differentiation of ROB cells.

Key words: naringin, miR-206, Cx43, ERK1, osteoblast, bone injury, drynariae, microRNAs

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