Chinese Journal of Tissue Engineering Research ›› 2026, Vol. 30 ›› Issue (36): 9381-9392.doi: 10.12307/2026.908

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Aucubin promotes osteogenic differentiation of mouse cranial pre-osteoblasts MC3T3-E1

Wang Houyuan1, 2, Xiao Jiacong1, 2, Shao Shoujia1, 3, He Paian1, 3, Yang Nan1, 3, Jiang Ziwei1, 2, 3   

  1. ¹Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China; 2The First Clinical Medical College of Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China; 3The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China
  • Received:2025-10-29 Revised:2026-03-14 Online:2026-12-28 Published:2026-05-20
  • Contact: Jiang Ziwei, PhD, Chief physician, Doctoral supervisor, Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China; The First Clinical Medical College of Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China; The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China
  • About author:Wang Houyuan, PhD candidate, Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China; The First Clinical Medical College of Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China
  • Supported by:
    National Natural Science Foundation of China, No. 81974575 (to JZW)

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Abstract: BACKGROUND: Oxidative stress is a major contributor to osteoporosis and other bone metabolic disorders. Aucubin, a natural compound with antioxidant and osteogenic effects, shows therapeutic potential, but its mechanisms remain unclear.
OBJECTIVE: To investigate the potential targets and pathways through which aucubin promotes osteogenic differentiation using network pharmacology combined with in vitro validation.
METHODS: Potential aucubin targets related to osteogenesis were identified from TCMSP and GeneCards databases, and a protein-protein interaction network with enrichment analysis was constructed to predict target pathways. An oxidative stress model of MC3T3-E1 cells was established using hydrogen peroxide, and the cells were assigned to control, model, low- and high-dose aucubin intervention, and Aucubin+JAK2/STAT3 inhibitor groups. Cell viability and malondialdehyde level were detected, and alkaline phosphatase staining and alizarin red staining were performed. Western blot and RT-qPCR were used to detect the expression of Janus kinase 2/signal transducer and activator of transcription 3 pathway-related proteins and osteogenic proteins and genes.
RESULTS AND CONCLUSION: (1) Network pharmacology predicted that aucubin functioned mainly via the Janus kinase 2/signal transducer and activator of transcription 3 pathway, involving key targets such as Janus kinase 2, signal transducer and activator of transcription 3, Janus kinase 1, interleukin 2, and matrix metalloproteinase 9. (2) In vitro experiments demonstrated that aucubin reduced oxidative stress, decreased malondialdehyde levels, enhanced alkaline phosphatase activity and mineralization, and upregulated the expression of pathway proteins as well as the gene and protein expression of osteogenic markers (Runt-related transcription factor 2, osteocalcin, osteopontin). These effects were partially reversed by the pathway inhibitor AG490. Taken together, these findings indicate that aucubin alleviates oxidative stress and promotes osteogenic differentiation of MC3T3-E1 cells through activation of the Janus kinase 2/signal transducer and activator of transcription 3 pathway.

Key words: aucubin, oxidative stress, osteogenic differentiation, osteoporosis, network pharmacology, Janus kinase 2/signal transducer and activator of transcription 3

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