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

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

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(LCMS/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CIAKI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 contrastinduced 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 reninangiotensin 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.