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中华危重症医学杂志(电子版) ›› 2020, Vol. 13 ›› Issue (06) : 406 -411. doi: 10.3877/cma.j.issn.1674-6880.2020.06.002

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论著

基于同位素标记技术筛选造影剂致急性肾损伤老年患者的尿液生物标志物
韩利娜1, 周芳芳1, 叶红华2, 沈根2, 罗群1,()   
  1. 1. 315010 浙江宁波,宁波市第二医院肾内科
    2. 315010 浙江宁波,宁波市第二医院心内科
  • 收稿日期:2020-10-25 出版日期:2020-12-31
  • 通信作者: 罗群
  • 基金资助:
    浙江省医药卫生科技项目(2013KYB236); 宁波市自然科学基金项目(2013A610266)

Screening for urine biomarkers in elderly patients with contrast-induced acute kidney injury based on isobaric tags for relative and absolute quantitation

Lina Han1, Fangfang Zhou1, Honghua Ye2, Gen Shen2, Qun Luo1,()   

  1. 1. Department of Nephrology, Ningbo No.2 Hospital, Ningbo 315010, China
    2. Department of Cardiology, Ningbo No.2 Hospital, Ningbo 315010, China
  • Received:2020-10-25 Published:2020-12-31
  • Corresponding author: Qun Luo
引用本文:

韩利娜, 周芳芳, 叶红华, 沈根, 罗群. 基于同位素标记技术筛选造影剂致急性肾损伤老年患者的尿液生物标志物[J/OL]. 中华危重症医学杂志(电子版), 2020, 13(06): 406-411.

Lina Han, Fangfang Zhou, Honghua Ye, Gen Shen, Qun Luo. Screening for urine biomarkers in elderly patients with contrast-induced acute kidney injury based on isobaric tags for relative and absolute quantitation[J/OL]. Chinese Journal of Critical Care Medicine(Electronic Edition), 2020, 13(06): 406-411.

目的

应用同位素标记相对和绝对定量(iTRAQ)技术联合液相色谱-串联质谱(LC-MS / MS)技术筛选造影剂致急性肾损伤(CI-AKI)老年患者早期诊断的尿液生物标记物。

方法

收集48例行经皮冠状动脉介入(PCI)术的老年患者一般资料,并将发生CI-AKI的患者纳入CI-AKI组(6例),同期从未发生CI-AKI的病例中按照1 ∶ 1比例选取患者纳入对照组(6例)。收集两组患者术后6 h尿液,并同时监测术前及术后24、48 h血肌酐水平。应用iTRAQ技术联合LC-MS / MS技术筛选出CI-AKI患者PCI术前后的相关差异蛋白,并进行生物信息学分析。

结果

两组患者在各时间点血肌酐水平比较,差异有统计学意义(F = 6.591,P = 0.015);CI-AKI组患者术后48 h的血肌酐水平较同组术前显著升高,且与对照组患者术后48 h比较亦显著升高[(133 ± 41)、(94 ± 34)、(81 ± 32)μmol / L,P均< 0.05]。与PCI术前相比,CI-AKI患者术后6 h共鉴定出差异蛋白151个,与CI-AKI相关的主要差异蛋白为血管紧张素原、人甘露糖结合凝集素相关丝氨酸蛋白酶2、免疫球蛋白G2重链、血清载脂蛋白A1、人过氧化物还原酶5,主要参与补体和凝血级联反应、肾素-血管紧张素系统通路等。

结论

iTRAQ技术能早期筛选出行PCI术后老年患者发生CI-AKI的潜在生物标志物,为临床早期诊断CI-AKI提供分子依据。

Objective

To identify primary urine biomarkers in elderly patients with contrast-induced acute kidney injury (CI-AKI) based on isobaric tags for relative and absolute quantitation (iTRAQ) and liquid chromatography tandem mass spectrometry (LC-MS / MS).

Methods

A total of 48 elderly patients undergoing percutaneous coronary intervention (PCI) were enrolled. Patients with CI-AKI were included in the CI-AKI group (n = 6), and patients without CI-AKI in the same period were selected as the control group in a 1 ∶ 1 ratio (n = 6). Their urine samples were collected at 6 h after PCI. Their serum creatinine levels were detected before PCI and 24 h, 48 h after PCI. The iTRAQ technique and LC-MS / MS were used to screen differentially expressed proteins of CI-AKI patients before and after PCI, and bioinformatic analysis was used to identify their potential biomarkers.

Results

The levels of serum creatinine showed significant differences between the two groups at different time points (F = 6.591, P = 0.015). The levels of serum creatinine in the CI-AKI group at 48 h after PCI were significantly higher than those in the same group before PCI, and higher than those in the control group at 48 h after PCI [(133 ± 41), (94 ± 34), (81 ± 32) μmol / L, both P < 0.05]. Compared with before PCI, 151 differentially expressed proteins were identified in CI-AKI patients at 6 h after PCI. The main differentially expressed proteins associated with CI-AKI were angiotensinogen, mannose binding lectin associated serine protease 2, immunoglobulin heavy G2 chain, apolipoprotein A1 and peroxiredoxin-5. They were mainly involved in the complement and coagulation cascade reaction and the renin-angiotensin system pathway.

Conclusion

The iTRAQ technology can screen potential biomarkers of CI-AKI in elderly patients undergoing PCI, and provide a molecular basis for early diagnosis of CI-AKI.

表1 两组行PCI治疗的老年患者一般资料的比较(± s
表2 各差异蛋白中生物学过程、细胞定位和分子功能排名前20的条目及所占比例(%)
生物学过程 细胞定位 分子功能
损伤反应(4) 细胞外膜(8) 结合蛋白(20)
防御反应(6) 细胞外泌体(8) 调节肽酶活性(3)
免疫过程(6) 细胞外细胞器(8) 肽链内切酶抑制剂活性(3)
单多细胞生物过程(9) 细胞外囊泡(8) 肽链内切酶活性(3)
应激反应(8) 细胞外区域部分(8) 肽酶抑制剂活性(3)
多细胞生物过程(10) 膜结合囊泡(8) 酶抑制剂活性(4)
生物调节(3) 囊泡(8) 结合细胞黏附分子(3)
蛋白激活级联瀑布反应(2) 细胞外区域(9) 丝氨酸型内肽酶抑制剂(2)
伤口愈合(3) 细胞外(5) 酶调节剂活性(5)
蛋白水解(6) 血液微粒(< 2) 结合受体(5)
肽酶活性调节(3) 膜结合细胞器(8) 结合糖胺聚糖(2)
创伤反应调节(3) 细胞器(9) 结合糖蛋白(< 2)
定位(10) 分泌颗粒(< 2) 结合蛋白酶(< 2)
炎症反应(3) 细胞质膜结合囊泡腔(< 2) 结合肝素(< 2)
凝血(3%) 囊泡内腔(< 2) 结合蛋白质复合物(4)
免疫反应(5) 胞浆囊泡(2) 结合脂肪酸(< 2)
细胞外基质形成(2) 细胞质膜结合囊泡(2) 抗氧化活性(< 2)
细胞外结构形成(2) 分泌颗粒内腔(< 2) 结合大分子复合物(5)
体液免疫反应(2) 血小板α颗粒管腔(< 2) 结合作用(24)
内肽酶活性的调节(< 2) 溶菌素(< 2) 结合整联蛋白(< 2)
表3 各差异蛋白参与的排名前10位信号通路
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