中国组织工程研究

中国组织工程研究 ›› 2025, Vol. 29 ›› Issue (5): 908-916.doi: 10.12307/2025.209

• 骨组织构建 bone tissue construction • 上一篇    下一篇

白藜芦醇激活细胞外信号调节激酶5信号蛋白促进小鼠MC3T3-E1细胞增殖

牛永康1，2，3，冯志尉1，2，3，王耀斌1，2，3，刘众成1，2，3，向德剑1，2，3，梁晓远1，2，3，移  植1，2，3，詹红伟1，2，3，耿  彬1，2，3，夏亚一1，2，3   

  1. 1兰州大学第二医院骨科，甘肃省兰州市  730030，2甘肃省骨科临床医学研究中心，甘肃省兰州市  730030，3甘肃省智能骨科行业技术中心，甘肃省兰州市  730030
  • 收稿日期:2023-12-25 接受日期:2024-01-31 出版日期:2025-02-18 发布日期:2024-06-01
  • 通讯作者: 夏亚一，博士，教授，主任医师，博士生导师，兰州大学第二医院骨科，甘肃省兰州市 730030；甘肃省骨科临床医学研究中心，甘肃省兰州市 730030；甘肃省智能骨科行业技术中心，甘肃省兰州市 730030
  • 作者简介:牛永康，男，1998年生，甘肃省天水市人，汉族，兰州大学在读硕士，主要从事成骨细胞、细胞生物力学的研究。
  • 基金资助:
    国家自然科学基金(81960403)，项目负责人：耿彬；国家自然科学基金(82060405，82360436)，项目负责人：夏亚一；甘肃省自然科学基金(22JR5RA943)，项目负责人：耿彬；甘肃省自然科学基金(22JR5RA956)，项目负责人：夏亚一

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)

PDF

468

摘要：




文题释义：
细胞外信号调节激酶5(extracellular signal regulated kinase 5，ERK5)：属于丝裂原活化蛋白激酶家族成员，由丝裂原活化蛋白激酶7基因编码。ERK5信号转导在许多生物过程中起着至关重要的作用，例如细胞的增殖和分化。
白藜芦醇(Resveratrol，RES)：是一种多酚，在葡萄皮和花生等植物中含量丰富，具有许多特性，包括抗糖化、抗氧化应激、抗炎和抗神经退行性变等，由于白藜芦醇通常耐受性良好，因此被认为是一种极具潜力的预防和治疗许多疾病的化合物。

背景：细胞外信号调节激酶5信号蛋白对生物体的存活不可或缺，白藜芦醇能通过多种途径促进成骨细胞增殖，但其是否能通过细胞外信号调节激酶5信号蛋白调控成骨细胞功能还需进一步验证。
目的：探究细胞外信号调节激酶5对MC3T3-E1细胞增殖以及相关分泌蛋白的调控作用，进一步验证白藜芦醇通过激活细胞外信号调节激酶5完成上述过程。
方法：小鼠MC3T3-E1前成骨细胞分别用完全培养基、XMD8-92(细胞外信号调节激酶5抑制剂)、表皮生长因子(细胞外信号调节激酶5激活剂)和白藜芦醇单独干预及XMD8-92+表皮生长因子、白藜芦醇+XMD8-92干预后，通过Western blot检测各组细胞内细胞外信号调节激酶5、磷酸化细胞外信号调节激酶5蛋白，增殖相关蛋白Cyclin D1、CDK4、PCNA，以及成骨细胞分泌蛋白骨保护素、核因子κB受体活化因子配体的表达情况，使用细胞免疫荧光染色检测各组细胞外信号调节激酶5、骨保护素和核因子κB受体活化因子配体荧光强度，使用EdU染色检测各组细胞增殖情况。白藜芦醇干预MC3T3-E1细胞的适宜浓度及时间由细胞形态学观察和CCK-8实验确定。
结果与结论：①细胞外信号调节激酶5信号蛋白的激活能有效促进MC3T3-E1细胞增殖、上调骨保护素/核因子κB受体活化因子配体比值；②白藜芦醇干预MC3T3-E1细胞的适宜浓度及时间为5 μmol/L，24 h；③白藜芦醇可以激活细胞外信号调节激酶5信号蛋白，进而促进成骨细胞增殖，并上调骨保护素/核因子κB受体活化因子配体比值；④研究结果表明，白藜芦醇可以通过激活细胞外信号调节激酶5信号蛋白促进MC3T3-E1细胞增殖，并通过激活细胞外信号调节激酶5信号蛋白上调骨保护素/核因子κB受体活化因子配体比值。  
https://orcid.org/0009-0005-7772-1142(牛永康)
中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: 细胞外信号调节激酶5, 白藜芦醇, 增殖, 骨保护素, 核因子κB受体活化因子配体

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

中图分类号: