UHPLC-MS/MS非靶向代谢组学在移居高原人群高原性高血压血浆代谢研究中的应用

王超臣; 唐才智; 冉庄; 罗勇军

doi:10.16016/j.2097-0927.202311001

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UHPLC-MS/MS非靶向代谢组学在移居高原人群高原性高血压血浆代谢研究中的应用

军事医学 | 更新时间：2026-01-09

- UHPLC-MS/MS非靶向代谢组学在移居高原人群高原性高血压血浆代谢研究中的应用
- Application of UHPLC-MS/MS-based nontargeted metabolomics in plasma metabolism in altitude-related hypertension among high-altitude migrants
- 陆军军医大学学报 2024年46卷第19期页码：2249-2258
- 作者机构：
  
  1.400038 重庆，陆军军医大学(第三军医大学)陆军卫勤训练基地军事医学地理学教研室
  2.430070 武汉，中部战区总医院干部病房三科
- 作者简介：
  
  [罗勇军，E-mail: luoyj@tmmu.edu.cn LUO Yongjun, E-mail: luoyj@tmmu.edu.cn
- 基金信息：
  
  国家自然基金面上项目(42377466);国家科技部第二次青藏高原综合科学考察研究专题(2019QZKK0607)
- DOI：10.16016/j.2097-0927.202311001
  中图分类号： R331.1;R394.2;R544.1
- 收稿：2023-11-01，
  
  修回：2024-05-18，
  
  纸质出版：2024-10-15
- 稿件说明：
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王超臣, 唐才智, 冉庄, 等. UHPLC-MS/MS非靶向代谢组学在移居高原人群高原性高血压血浆代谢研究中的应用[J]. 陆军军医大学学报, 2024,46(19):2249-2258.

Chaocheng WANG, Caizhi TANG, Zhuang RAN, et al. Application of UHPLC-MS/MS-based nontargeted metabolomics in plasma metabolism in altitude-related hypertension among high-altitude migrants[J]. Journal of Army Medical University, 2024, 46(19): 2249-2258.
王超臣, 唐才智, 冉庄, 等. UHPLC-MS/MS非靶向代谢组学在移居高原人群高原性高血压血浆代谢研究中的应用[J]. 陆军军医大学学报, 2024,46(19):2249-2258. DOI： 10.16016/j.2097-0927.202311001.

Chaocheng WANG, Caizhi TANG, Zhuang RAN, et al. Application of UHPLC-MS/MS-based nontargeted metabolomics in plasma metabolism in altitude-related hypertension among high-altitude migrants[J]. Journal of Army Medical University, 2024, 46(19): 2249-2258. DOI： 10.16016/j.2097-0927.202311001.

摘要

目的

研究高原性高血压(altitude-related hypertension

ARH)患者和健康个体之间血浆代谢产物的差异，并分析ARH的潜在发病机制。

方法

选取2020年7月在进驻海拔4 200 m处的健康平原成年男性官兵中按简单随机抽样法选取20名ARH患者为ARH组，30名血压正常者为对照(Control)组。在测量血压、体质指数(body mass index

BMI)、血氧饱和度和心率后，采集研究对象空腹静脉血样本。基于超高效液相色谱-质谱联用(ultra-high performance liquid chromatography-tandem mass spectrometry

UHPLC-MS/MS)技术，对研究对象血浆中的代谢产物进行筛选和鉴定。使用无监督主成分分析(principal component analysis

PCA)和有监督的正交偏最小二乘判别分析(orthogonal partial least squares discrimination analysis

OPLS-DA)模型对代谢产物进行指纹图谱分析，以辅助生物标志物的筛选，并对OPLS-DA模型进行质量考查和验证，以确保模型的稳定性和可靠性。使用独立样本

检验和差异倍数(fold change

FC)分析，绘制火山图，筛选有差异的血浆代谢物。使用京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes，KEGG)富集通路分析对筛选出的差异性代谢物进行功能途径的富集分析和拓扑学分析。

结果

与Control组比较，ARH组的收缩压、舒张压和心率显著升高，动脉血氧饱和度显著降低(

＜0.05)。PCA分析显示，正离子模式下，解释了81.96%的变异；负离子模式下，解释了79.25%的变异，表明2组之间存在显著的代谢差异。OPLS-DA模型分析显示，正离子模式下，PC1解释了77.36%的变异，PC2解释了12.25%的变异，R

Y=0.96，Q

Y=0.91；负离子模式下，PC1解释了84.15%的变异，PC2解释了17.24%的变异，R

Y=0.99，Q

Y=0.86。组间差异均超过75%，组内差异均小于20%。正离子模式下，R

拟合直线的Y轴截距为0.58，Q

拟合直线的Y轴截距为-0.48；负离子模式下，R

拟合直线的Y轴截距为0.93，Q

拟合直线的Y轴截距为-0.41。共筛选出32种具有显著差异的代谢产物，包括氨基酸、核苷、脂肪酸和有机生物碱。KEGG分析显示，10条代谢通路中有4条为氨基酸代谢通路，氨酰-tRNA生物合成途径富集的代谢产物最多。

结论

基于UHPLC-MS/MS技术，运用非靶向代谢组学筛选出的32种具有明显差异的代谢产物可能作为ARH的特征性生物标志物，而氨酰-tRNA生物合成途径可能与ARH的发病机制相关。

Abstract

Objective

To investigate the differences in plasma metabolites between patients with altitude-related hypertension (ARH) and healthy individuals

and analyze the potential pathogenesis of ARH.

Methods

Convenient sampling was conducted on a unit of male healthy officers and soldiers who resident at altitude of < 500 m and migrated to an altitude of 4 200 m in July 2020. Twenty of them diagnosed with ARH were assigned into the ARH group

and another 30 non-ARH individuals served as the control group. Their blood pressure

body mass index (BMI)

blood oxygen saturation

and heart rate were measured and recorded

and fasting venous blood samples were harvested to screen and identify plasma metabolites with ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Metabolite fingerprinting was performed using unsupervised Principal Component Analysis (PCA) and supervised Orthogonal Partial Least Squares Discrimination Analysis (OPLS-DA) models in order to assist in biomarker screening. The quality of the OPLS-DA model was assessed and validated to guarantee the stability and reliability of the model. Differential plasma metabolites were screened using independent sample t test and fold change (FC) analysis

and volcano plots were drawn. Finally

Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment pathway analysis was used to perform functional pathway enrichment and topological analysis on the screened differential metabolites.

Results

Compared to the control group

the ARH group showed significantly higher systolic and diastolic blood pressure and heart rate

and lower arterial oxygen saturation (

0.05). PCA analysis showed that 81.96% of the variance was explained in the po

sitive ion mode and 79.25% in the negative ion mode

indicating significant metabolic differences between the 2 groups. OPLS-DA model analysis indicated that in the positive ion mode

PC1 explained 77.36% of the variance and PC2 explained 12.25% of the variance

with R

Y=0.96 and Q

Y=0.91; in the negative ion mode

PC1 explained 84.15% of the variance and PC2 explained 17.24% of the variance

with R

Y=0.99 and Q

Y=0.86. Inter-group difference exceeded 75%

and intra-group difference was less than 20%. The 7-fold cross-validation and 200 permutation test confirmed that the model was stable and reliable. In the positive ion mode

the Y-axis intercepts of the R

and Q

fitted lines were 0.58 and -0.48

respectively; in the negative ion mode

the Y-axis intercepts were 0.93 and -0.41

respectively. A total of 32 significantly different metabolites were screened out

including amino acids

nucleosides

fatty acids

and organic alkaloids. KEGG analysis revealed that among the 10 metabolic pathways

4 were amino acid metabolic pathways

with the aminoacyl-tRNA biosynthesis pathway having the most enriched metabolites.

Conclusion

Based on UHPLC-MS/MS technology

untargeted metabolomics analysis identifies 32 significantly different metabolites

which may serve as characteristic biomarkers for ARH

and the aminoacyl-tRNA biosynthesis pathway may be associated with the pathogenesis of ARH.

关键词

Keywords

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