Boyd J Gordon, Smithson Laura J, Howes Daniel, Muscedere John, Kawaja Michael D
Department of Critical Care Medicine, Queen's University/Kingston General Hospital, Kingston, Ontario, Canada.
Queen's Centre for Neuroscience Studies, Queen's University, 2nd Floor, Botterell Hall, Kingston, Ontario, Canada.
Intensive Care Med Exp. 2016 Dec;4(1):9. doi: 10.1186/s40635-016-0084-3. Epub 2016 May 10.
Serum biomarkers may play a role in prognostication after cardiac arrest. This study was designed to assess the feasibility of using two-dimensional gel electrophoresis (2D-GE) coupled with mass spectrometry (MS) as a proteomic strategy to identify novel biomarkers that may predict neurological recovery.
Adult comatose survivors of ventricular fibrillation or pulseless ventricular tachycardia were considered eligible. Blood was collected and serum separated within 6 h of hospital admission and then at 24 h afterwards. Neurological outcome was assessed at 3 months with the Cerebral Performance Category (CPC) score. Serum was assessed with 2D-GE with and without prior depletion of high abundance proteins. Protein differences between patients with good (CPC 1,2) vs. poor (CPC 3-5) neurological recovery were subsequently identified with MS.
From August 2010 to June 2014, 11 patients meeting eligibility criteria were recruited, from which serum was available from 9 (5 with good neurological outcome). On non-depleted serum, only high abundance acute phase proteins such as haptoglobin, cell-free hemoglobin, albumin, and amyloid were detected in both patients with good and poor neurological recovery. Following depletion of high abundance proteins, proteins identified by MS in both patient populations were the acute phase reactants c-reactive protein and retinol binding protein-4. Proteins uniquely identified in the serum of patients with poor neurological recovery included 14-3-3 (epsilon and zeta isoforms) and muskelin.
Two-D-GE coupled with MS is a feasible strategy to facilitate the identification of novel predictive biomarkers. The presence of muskelin and 14-3-3 in the serum of patients with poor neurological prognosis warrants further investigation.
血清生物标志物可能在心脏骤停后的预后评估中发挥作用。本研究旨在评估二维凝胶电泳（2D-GE）结合质谱（MS）作为一种蛋白质组学策略来识别可能预测神经功能恢复的新型生物标志物的可行性。
心室颤动或无脉性室性心动过速的成年昏迷幸存者被认为符合条件。在入院6小时内及之后24小时采集血液并分离血清。在3个月时用脑功能分级（CPC）评分评估神经功能结局。对血清进行2D-GE分析，分别采用有无预先去除高丰度蛋白的方法。随后用质谱鉴定神经功能恢复良好（CPC 1、2）与恢复较差（CPC 3-5）患者之间的蛋白质差异。
从2010年8月至2014年6月，招募了11名符合入选标准的患者，其中9名患者有可用血清（5名神经功能结局良好）。在未去除高丰度蛋白的血清中，神经功能恢复良好和恢复较差的患者均仅检测到高丰度急性期蛋白，如触珠蛋白、游离血红蛋白、白蛋白和淀粉样蛋白。去除高丰度蛋白后，两组患者经质谱鉴定的蛋白均为急性期反应物C反应蛋白和视黄醇结合蛋白-4。在神经功能恢复较差患者血清中独特鉴定出的蛋白包括14-3-3（ε和ζ亚型）和肌动蛋白。
二维凝胶电泳结合质谱是促进鉴定新型预测性生物标志物的可行策略。神经功能预后较差患者血清中存在肌动蛋白和14-3-3值得进一步研究。
