中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2021, Vol. 16 ›› Issue (7): 1294-1301.doi: 10.4103/1673-5374.301487

• 原著:脊髓损伤修复保护与再生 • 上一篇    下一篇

质谱法鉴定红细胞中潜在的脊髓损伤氧化应激生物标志物

  


  • 出版日期:2021-07-15 发布日期:2021-01-07
  • 基金资助:

    宁夏回族自治区重点科研项目（2018BCG01002）；宁夏2017年学科建设项目研究生教育创新计划（YXW2017014）

Identification of potential oxidative stress biomarkers for spinal cord injury in erythrocytes using mass spectrometry

Li-Jian Zhang1, 2, 3, #, Yao Chen1, 2, 3, #, Lu-Xuan Wang1, Xiao-Qing Zhuang4, *, He-Chun Xia2, 3, *   

  1. 1 School of Clinical Medicine, Ningxia Medical University, Yinchuan, Ningxia Hui Autonomous Region, China;  2 Department of Neurosurgery, General Hospital of Ningxia Medical University, Yinchuan, Ningxia Hui Autonomous Region, China;  3 Ningxia Human Stem Cell Research Institute, General Hospital of Ningxia Medical University, Yinchuan, Ningxia Hui Autonomous Region, China;  4 Department of Nuclear Medicine, General Hospital of Ningxia Medical University, Yinchuan, Ningxia Hui Autonomous Region, China

  • Online:2021-07-15 Published:2021-01-07
  • Contact: He-Chun Xia, MD, xhechun@nyfy.com.cn; Xiao-Qing Zhuang, PhD, zhuangxq@nyfy.com.cn.
  • Supported by:
    This work was supported by the Key Research Projects of the Ningxia Hui Autonomous Region of China, No. 2018BCG01002 (to HCX); and the Plan of Postgraduate Education Innovation, Discipline Construction Project of Ningxia, China (2017), No. YXW2017014 (to LJZ).

PDF

114

摘要：

氧化应激是脊髓损伤继发性损伤的标志。寻找稳定敏感的氧化应激生物学标记物对研究脊髓损伤继发性损伤具有重要意义。成熟红细胞不含有细胞核和线粒体，不能进行蛋白转录和翻译，因而对于氧化应激高度敏感，可能成为一种有价值的生物标志物。实验以比格犬脊髓半横断损伤模型为对象，应用质谱蛋白组学分析首次揭示脊髓损伤急性和亚急性期红细胞中蛋白表达的动态变化特点。结果共检出26种蛋白可在脊髓损伤后急性(0-3d)和亚急性期(7-21d)差异表达。进一步生物信息学分析表明，这些差异表达的蛋白参与谷胱甘肽代谢、脂质代谢和戊糖磷酸通路等氧化应激过程。western blot验证了谷胱甘肽合成酶、转醛基酶和髓过氧化物酶3种差异表达蛋白的表达情况，结果与质谱结果一致。表明红细胞可作为脊髓损伤氧化应激生物学标记物的一种新颖的样本来源。来源于红细胞的谷胱甘肽合成酶、转醛基酶和髓过氧化物酶是脊髓损伤后氧化应激的潜在标志物。动物实验于2017年2月13日获得宁夏医科大学实验动物中心批准，批准号：2017-073。

https://orcid.org/0000-0002-3058-8162 (He-Chun Xia); 

https://orcid.org/0000-0001-8652-8540 (Xiao-Qing Zhuang);

https://orcid.org/0000-0003-3881-7705 (Li-Jian Zhang); 

https://orcid.org/0000-0001-9071-2578 (Lu-Xuan Wang)

关键词:

中枢神经, 脊髓损伤, 犬, 急性期, 亚急性期, 外周血, 红细胞, 质谱, 生物信息学, 氧化应激, 生物标志物

Abstract: Oxidative stress is a hallmark of secondary injury associated with spinal cord injury. Identifying stable and specific oxidative biomarkers is of important significance for studying spinal cord injury-associated secondary injury. Mature erythrocytes do not contain nuclei and mitochondria and cannot be transcribed and translated. Therefore, mature erythrocytes are highly sensitive to oxidative stress and may become a valuable biomarker. In the present study, we revealed the proteome dynamics of protein expression in erythrocytes of beagle dogs in the acute and subacute phases of spinal cord injury using mass spectrometry-based approaches. We found 26 proteins that were differentially expressed in the acute (0–3 days) and subacute (7–21 days) phases of spinal cord injury. Bioinformatics analysis revealed that these differentially expressed proteins were involved in glutathione metabolism, lipid metabolism, and pentose phosphate and other oxidative stress pathways. Western blot assays validated the differential expression of glutathione synthetase, transaldolase, and myeloperoxidase. This result was consistent with mass spectrometry results, suggesting that erythrocytes can be used as a novel sample source of biological markers of oxidative stress in spinal cord injury. Glutathione synthetase, transaldolase, and myeloperoxidase sourced from erythrocytes are potential biomarkers of oxidative stress after spinal cord injury. This study was approved by the Experimental Animal Centre of Ningxia Medical University, China (approval No. 2017-073) on February 13, 2017.

Key words: acute phase, bioinformatic analysis, biomarkers, central nervous system, dog, erythrocytes, mass spectrometry, oxidative stress, peripheral blood, spinal cord injury, subacute phase