Chinese Journal of Tissue Engineering Research ›› 2025, Vol. 29 ›› Issue (5): 908-916.doi: 10.12307/2025.209

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Resveratrol activates extracellular-regulated protein kinase 5 signaling protein to promote proliferation of mouse MC3T3-E1 cells 

Niu Yongkang1, 2, 3, Feng Zhiwei1, 2, 3, Wang Yaobin1, 2, 3, Liu Zhongcheng1, 2, 3, Xiang Dejian1, 2, 3, Liang Xiaoyuan1, 2, 3, Yi Zhi1, 2, 3, Zhan Hongwei1, 2, 3, #br# Geng Bin1, 2, 3, Xia Yayi1, 2, 3 #br#   

  1. 1Department of Orthopedics, Lanzhou University Second Hospital, Lanzhou 730030, Gansu Province, China; 2Orthopedic Clinical Research Center of Gansu Province, Lanzhou 730030, Gansu Province, China; 3Intelligent Orthopedic Industry Technology Center of Gansu Province, Lanzhou 730030, Gansu Province, China 
  • Received:2023-12-25 Accepted:2024-01-31 Online:2025-02-18 Published:2024-06-01
  • Contact: Xia Yayi, MD, Professor, Chief physician, Doctoral supervisor, Department of Orthopedics, Lanzhou University Second Hospital, Lanzhou 730030, Gansu Province, China; Orthopedic Clinical Research Center of Gansu Province, Lanzhou 730030, Gansu Province, China; Intelligent Orthopedic Industry Technology Center of Gansu Province, Lanzhou 730030, Gansu Province, China
  • About author:Niu Yongkang, Master candidate, Department of Orthopedics, Lanzhou University Second Hospital, Lanzhou 730030, Gansu Province, China; Orthopedic Clinical Research Center of Gansu Province, Lanzhou 730030, Gansu Province, China; Intelligent Orthopedic Industry Technology Center of Gansu Province, Lanzhou 730030, Gansu Province, China
  • Supported by:
    the National Natural Science Foundation of China, No. 81960403 (to GB); the National Natural Science Foundation of China, Nos. 82060405 and 82360436 (to XYY); the Natural Science Foundation of Gansu Province, No. 22JR5RA943 (to GB); the Natural Science Foundation of Gansu Province, No. 22JR5RA956 (to XYY)

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Abstract: BACKGROUND: The extracellular-regulated protein kinase 5 (ERK5) signaling protein is essential for the survival of organisms, and resveratrol can promote osteoblast proliferation through various pathways. However, whether resveratrol can regulate osteoblast function through the ERK5 signaling protein needs further verification.
OBJECTIVE: To explore the regulatory effect of ERK5 on the proliferation of MC3T3-E1 cells and related secreted proteins, and to further verify whether resveratrol can complete the above process by activating ERK5.
METHODS: Mouse MC3T3-E1 preosteoblasts were treated with complete culture medium, XMD8-92 (an ERK5 inhibitor), epidermal growth factor (an ERK5 activator), resveratrol alone, XMD8-92+EGF, and resveratrol+XMD8-92, respectively. Western blot assay was used to detect the expression of ERK5 and p-ERK5 proteins, proliferation-related proteins Cyclin D1, CDK4 and PCNA, and osteoblast-secreted proteins osteoprotegerin and receptor activator of nuclear factor-κB ligand in MC3T3-E1 cells of each group. The fluorescence intensity of ERK5, osteoprotegerin and receptor activator of nuclear factor-κB ligand in each group was detected by cell immunofluorescence staining, and cell proliferation was detected by EdU staining, respectively. The appropriate concentration and time of resveratrol intervention in MC3T3-E1 cells were determined by cell morphology observation and cell counting kit-8 assay.
RESULTS AND CONCLUSION: The activation of ERK5 signaling protein could effectively promote the proliferation of MC3T3-E1 cells, up-regulate the osteoprotegerin/receptor activator of nuclear factor-κB ligand ratio. The appropriate concentration and time for resveratrol intervention in MC3T3-E1 cells was 5 μmol/L and 24 hours, respectively. Resveratrol could activate ERK5 signaling protein, thereby promoting osteoblast proliferation and up-regulating the osteoprotegerin/RANKL ratio. All these results indicate that resveratrol can promote the proliferation of MC3T3-E1 cells and up-regulate the osteoprotegerin/RANKL ratio by activating the ERK5 signaling protein.

Key words:  extracellular signal-regulated kinase 5, resveratrol, proliferation, osteoprotegerin, receptor activator of nuclear factor-kappa B ligand

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