中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (36): 9393-9401.doi: 10.12307/2026.381

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

高糖诱导成骨细胞铜死亡的机制

连  勇1，谢志兴1，陆嘉欣1，潘兆丰1，陈庆真2，邵  敏2   

  1. 1广州中医药大学第三临床医学院，广东省广州市   510405；2广州中医药大学第三附属医院，广东省广州市   510405
  • 收稿日期:2025-07-06 修回日期:2025-09-19 出版日期:2026-12-28 发布日期:2026-05-20
  • 通讯作者: 邵敏，博士，主任医师，硕士/博士生导师，广州中医药大学第三附属医院，广东省广州市 510240
  • 作者简介:连勇，男，1990年生，山东省枣庄市人，广州中医药大学博士三年级在读，主要从事骨质疏松和骨关节病的研究。
  • 基金资助:
    广东省普通高校特色创新项目(2024KTSCX115)，项目负责人：陈庆真

Mechanism of high glucose-induced osteoblast cuproptosis

Lian Yong1, Xie Zhixing1, Lu Jiaxin1, Pan Zhaofeng1, Chen Qingzhen2, Shao Min2   

  1. 1The Third Clinical Medical College of Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China; 2The Third Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China
  • Received:2025-07-06 Revised:2025-09-19 Online:2026-12-28 Published:2026-05-20
  • Contact: Shao Min, MD, Chief physician, Master’s supervisor, Doctoral supervisor, the Third Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China
  • About author:Lian Yong, MD candidate, the Third Clinical Medical College of Guangzhou University of Chinese Medicine, Guangzhou 510405, Guangdong Province, China
  • Supported by:
    Guangdong Provincial Special Innovation Project for Ordinary Higher Education Institutions, No. 2024KTSCX115 (to CQZ)

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摘要：



文题释义：
铜死亡：一种新型的铜依赖性细胞死亡形式，过量的铜通过促进依赖铜的脂酰化蛋白在三羧酸循环中异常寡聚化以及降低Fe-S簇蛋白水平来诱导铜死亡。
溶质载体家族31成员1/铁氧还蛋白1(SLC31A1/FDX1)轴：为铜死亡的关键调控通路，SLC31A1负责铜离子转运，FDX1参与铁硫簇蛋白调控。

摘要
背景：铜死亡是一种新型细胞死亡方式，近年来在多种疾病的发生发展中受到广泛关注，其中铜死亡机制在肿瘤领域已有研究，但铜死亡对成骨细胞的影响尚未明确。
目的：探讨高糖诱导下成骨细胞铜死亡的机制。
方法：①将MC3T3-E1细胞分4组培养，分别为20 μmol/L CuCl2组、40 μmol/L CuCl2组、20 μmol/L CuCl2+高糖组、40 μmol/L CuCl2+高糖组，其中加入25 mmol/L葡萄糖+200 mmol/L棕榈酸钠模拟高糖环境。成骨诱导培养48 h后，Western Blot检测铜离子转运酶(铜离子转入酶SLC31A1、铜离子转出酶ATP7B)的蛋白表达。②将MC3T3-E1细胞分4组培养：正常组不进行任何处理，高糖组加入25 mmol/L葡萄糖与200 mmol/L棕榈酸钠，CuCl2组加入20 μmol/L CuCl2，高糖+CuCl2组同时加入25 mmol/L葡萄糖、200 mmol/L棕榈酸钠与20 μmol/L CuCl2。成骨诱导培养后，透射电镜下观察线粒体结构，碱性磷酸酶染色与茜素红染色检测成骨分化与矿化，qRT-PCR检测成骨特异性基因表达，Western Blot检测成骨与铜死亡相关蛋白表达。
结果与结论：①Western Blot检测显示，20 μmol/L CuCl2+高糖组SLC31A1蛋白表达高于20 μmol/L CuCl2组(P < 0.05)，ATP7B蛋白表达低于20 μmol/L CuCl2组(P < 0.05)；40 μmol/L CuCl2+高糖组SLC31A1蛋白表达高于40 μmol/L CuCl2组(P < 0.05)，ATP7B蛋白表达低于40 μmol/L CuCl2组(P < 0.05)。②透射电镜下可见CuCl2诱导了MC3T3-E1细胞线粒体结构改变，主要表现为线粒体嵴消失、线粒体皱缩、体积变小、线粒体膜结构破坏，联合高糖环境会加重这些结构的恶变。碱性磷酸酶与茜素红染色结果显示，CuCl2抑制了MC3T3-E1细胞的成骨分化与矿化，而CuCl2+高糖环境抑制MC3T3-E1细胞成骨分化与矿化的作用强于单独CuCl2。qRT-PCR与Western Blot检测结果显示，CuCl2+高糖环境可显著抑制成骨特异性基因mRNA和蛋白的表达，诱导成骨细胞铜死亡。③结果表明，高糖环境可能通过影响铜离子转运和铜死亡相关蛋白的表达诱导并加重成骨细胞铜死亡。

https://orcid.org/0009-0009-4097-927X (连勇)


中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 铜死亡, 成骨细胞, 高糖, SLC31A1/FDX1轴, MC3T3-E1细胞, 铜离子转运

Abstract: BACKGROUND: Cuproptosis is a newly identified copper-dependent cell death. It has attracted more attention to various diseases in recent years. Cuproptosis has been studied in cancer, but its role in osteoblasts remains unclear.
OBJECTIVE: To investigate the mechanism of high glucose-induced cuproptosis in osteoblasts.
METHODS: (1) MC3T3-E1 cells were cultured in four groups: 20 μmol/L CuCl2 group, 40 μmol/L CuCl2 group, 20 μmol/L CuCl2+high glucose group, and 40 μmol/L CuCl2+high glucose group. The high glucose environment was simulated by adding 25 mmol/L glucose and 200 mmol/L sodium palmitate. After 48 hours of osteogenic induction, western blot assay was used to detect the protein expression of copper ion transporters (copper ion influx transporter SLC31A1 and copper ion efflux transporter ATP7B). (2) MC3T3-E1 cells were cultured in four groups: the normal group without any treatment, the high glucose group with the addition of 25 mmol/L glucose and 200 mmol/L sodium palmitate, the CuCl2 group with the addition of 20 μmol/L CuCl2, and the high glucose + CuCl2 group with the simultaneous addition of 25 mmol/L glucose, 200 mmol/L sodium palmitate, and 20 μmol/L CuCl2. After osteogenic induction, mitochondrial alterations were observed via transmission electron microscopy. The osteoblast differentiation and mineralization were evaluated using alkaline phosphatase staining and alizarin red staining. qRT-PCR was used to detect osteoblast-specific gene expression, and western blot was used to detect osteoblast- and cuproptosis-related protein expression.
RESULTS AND CONCLUSION: (1) Western blot analysis showed that the expression of SLC31A1 protein was higher in the 20 μmol/L CuCl2+high glucose group than the 20 μmol/L CuCl2 group (P < 0.05), while the expression of ATP7B protein was lower (P < 0.05). In the 40 μmol/L CuCl2+high glucose group, the expression of SLC31A1 protein was higher than in the 40 μmol/L CuCl2 group (P < 0.05), and the expression of ATP7B protein was lower (P < 0.05). (2) Under the transmission electron microscopy, CuCl2 induced changes in the mitochondrial structure of MC3T3-E1 cells, primarily characterized by the disappearance of mitochondrial cristae, mitochondrial shrinkage, reduced volume, and disruption of mitochondrial membrane structure. The combined exposure to high glucose exacerbated these structural alterations. Alkaline phosphatase and alizarin red staining results indicated that CuCl2 inhibited osteogenic differentiation and mineralization of MC3T3-E1 cells, and the inhibitory effect of CuCl2+high glucose environment on osteogenic differentiation and mineralization was stronger than that of CuCl2 alone. qRT-PCR and western blot analysis revealed that the CuCl2+high glucose environment significantly inhibited the expression of osteogenic-specific gene at mRNA and protein levels, inducing osteoblast cuproptosis. These findings suggest that high glucose may induce and exacerbate osteoblast cuproptosis by disrupting copper transport and cuproptosis-related protein expression. 

Key words: cuproptosis, osteoblasts, high glucose, SLC31A1/FDX1 axis, MC3T3-E1 cell, copper transporter 

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