中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (33): 8645-8652.doi: 10.12307/2026.490

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同位素相对标记与绝对定量技术鉴定脊髓型颈椎病患者血清生物学标志物

许  伟1，崔子正2，胡明告3，刘春志2，李智斐1，钟远鸣1，杨汉立2   

  1. 1广西中医药大学第一附属医院，广西壮族自治区南宁市   530023；2广西中医药大学，广西壮族自治区南宁市   530001；3广东中能建电力医院，广东省广州市   510735
  • 收稿日期:2025-10-29 修回日期:2026-03-18 出版日期:2026-11-28 发布日期:2026-06-10
  • 通讯作者: 杨汉立，在读博士，广西中医药大学，广西壮族自治区南宁市 530001
  • 作者简介:许伟，博士，主治医师，广西中医药大学第一附属医院，广西壮族自治区南宁市 530023
  • 基金资助:
    国家自然科学基金项目(82260942)，项目负责人：钟远鸣；广西中医药大学校级课题项目(XP023091) ，项目负责人：杨汉立；广西中医药大学第一附属医院院级课题项目(院字【2023】29号)，项目负责人：许伟

Application of isobaric tags for relative and absolute quantification for identifying serum biomarkers in patients with cervical spinal cord syndrome

Xu Wei1, Cui Zizheng2, Hu Minggao3, Liu Chunzhi2, Li Zhifei1, Zhong Yuanming1, Yang Hanli2   

  1. 1The First Affiliated Hospital of Guangxi University of Chinese Medicine, Nanning 530023, Guangxi Zhuang Autonomous Region, China; 2Guangxi University of Chinese Medicine, Nanning 530001, Guangxi Zhuang Autonomous Region, China; 3Guangdong Zhongnengjian Electric Power Hospital, Guangzhou 510735, Guangdong Province, China
  • Received:2025-10-29 Revised:2026-03-18 Online:2026-11-28 Published:2026-06-10
  • Contact: Yang Hanli, PhD candidate, Guangxi University of Chinese Medicine, Nanning 530001, Guangxi Zhuang Autonomous Region, China
  • About author:Xu Wei, PhD, Attending physician, The First Affiliated Hospital of Guangxi University of Chinese Medicine, Nanning 530023, Guangxi Zhuang Autonomous Region, China
  • Supported by:
    National Natural Science Foundation of China, No. 82260942 (to ZYM); University-level Project of Guangxi University of Chinese Medicine, No. XP023091 (to YHL); Hospital-level Project of the First Affiliated Hospital of Guangxi University of Chinese Medicine, No. [2023] 29 (to XW) 

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摘要：


文题释义：
同位素相对标记与绝对定量技术：一种蛋白质组学技术，通过用不同质量的同位素标签标记不同样本的蛋白质，经混合后进行质谱分析，从而精准鉴定并定量蛋白质在不同样本中的表达差异。
脊髓型颈椎病：因颈椎间盘突出、骨质增生等结构病变压迫颈部脊髓，导致四肢麻木、行走不稳及精细动作障碍的严重神经系统疾病。

背景：脊髓型颈椎病是颈椎病中最严重的一种类型，但它与健康人群的特异性血清生物标志物尚不清楚。
目的：采用同位素相对标记与绝对定量技术分析气虚血瘀证脊髓型颈椎病患者与健康人群血清差异蛋白表达状况，筛选并确定该病症潜在的血清生物学标志物。
方法：研究分别纳入13例气虚血瘀证脊髓型颈椎病患者与8例健康受试者，采集血清样本运用同位素相对标记与绝对定量技术标记联合液相色谱-质谱联用技术进行蛋白质组学分析，通过生物信息学方法对差异蛋白进行功能注释与通路富集分析，揭示其可能的生物学意义和参与的信号通路。
结果与结论：①质谱分析共鉴定出78个差异表达蛋白(P < 0.05)，与健康人群相比，血管扩张刺激磷酸蛋白、多表皮生长因子样结构域蛋白9、补体因子H相关蛋白3及丝裂蛋白A等12种差异蛋白明显上调，转铁蛋白、软骨中间层蛋白2、纤维连接蛋白及纤维蛋白原β链等66种差异蛋白明显下调；②这些差异蛋白主要参与了核酸转运、蛋白质活化级联等多种生物过程和具有与蛋白激酶B结合、免疫球蛋白受体结合及翻译延伸因子活性等多种分子功能，涉及矿物质吸收、氧化磷酸化、血小板活化等7条信号通路；③气虚血瘀证脊髓型颈椎病患者与健康人群血清差异蛋白中IGLC2、IGLC7、IGLL5、IGLV2-8、IGLV3-25、IGKV3D-11等在功能网络中与其他蛋白广泛相互作用；④使用Western Blot法对差异蛋白IGLC2验证，其结果与蛋白质组学分析结果相符；⑤研究结果表明，同位素相对标记与绝对定量技术联合液相色谱-质谱联用技术成功筛选出气虚血瘀证脊髓型颈椎病人群与健康人群差异蛋白，并初步推测IGLC2、IGLL5及IGLV3-25等7种蛋白是该人群的特异性血清标志物。
https://orcid.org/0009-0002-6666-534X(杨汉立) 


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: 脊髓型颈椎病, 蛋白质组学, 同位素相对标记与绝对定量技术, 血清标志物, 气虚血瘀证

Abstract: BACKGROUND: Cervical spondylotic myelopathy is the most severe type of cervical spondylosis, yet its specific serum biomarkers that can distinguish it from healthy individuals remain unclear.  
OBJECTIVE: To analyze the differential expression of serum proteins between patients with cervical spondylotic myelopathy of Qi deficiency and blood stasis type and healthy individuals using isobaric tags for relative and absolute quantification (iTRAQ), and to screen and identify potential serum biomarkers for this condition.  
METHODS: Serum samples were collected from 13 patients with cervical spondylotic myelopathy of Qi deficiency and blood stasis type and 8 healthy individuals. Proteomic analysis was performed using iTRAQ with liquid chromatography-tandem mass spectrometry. Functional annotation and pathway enrichment analysis of differentially expressed proteins were conducted using bioinformatics methods to reveal their potential biological significance and involved signaling pathways.  
RESULTS AND CONCLUSION: (1) Mass spectrometry analysis identified a total of 78 differentially expressed proteins (P < 0.05). Compared with healthy individuals, 12 differential proteins, including vasodilator-stimulated phosphoprotein, multiple epidermal growth factor-like domains protein 9, complement factor H-related protein 3, and mitogenin A, were significantly upregulated, while 66 differential proteins, including transferrin, cartilage intermediate layer protein 2, fibronectin, and fibrinogen beta chain, were significantly downregulated. (2) These differentially expressed proteins were primarily involved in various biological processes such as nucleic acid transport and protein activation cascade, and exhibited molecular functions including protein kinase B binding, immunoglobulin receptor binding, and translation elongation factor activity. They were associated with seven signaling pathways, including mineral absorption, oxidative phosphorylation, and platelet activation. (3) Among the differentially expressed proteins in the serum of patients with cervical spondylotic myelopathy of Qi deficiency and blood stasis type compared with healthy individuals, IGLC2, IGLC7, IGLL5, IGLV2-8, IGLV3-25, and IGKV3D-11 showed extensive interactions with other proteins in the functional network. (4) Validation of the differentially expressed protein IGLC2 using western blot yielded results consistent with the proteomics analysis. To conclude, these findings indicate that iTRAQ combined with liquid chromatography-tandem mass spectrometry successfully screened differentially expressed proteins between patients with cervical spondylotic myelopathy of Qi deficiency and blood stasis type and healthy individuals, and preliminarily suggest that seven proteins, including IGLC2, IGLL5, and IGLV3-25, may serve as specific serum biomarkers for this population.  


Key words: cervical spondylotic myelopathy, proteomics, isobaric tags for relative and absolute quantification, serum biomarkers, Qi deficiency and blood stasis 

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