Gong Yu, Chen Nan, Wang Fu-Qiang, Wang Zhao-Hui, Xu Hua-Xi
Department of Nephrology, Affiliated Renmin Hospital, School of Medical Science and Laboratory Medicine, Jiangsu University, Zhenjiang, China.
Nephrol Dial Transplant. 2009 Oct;24(10):3108-14. doi: 10.1093/ndt/gfp231. Epub 2009 May 21.
Continuous renal replacement therapy (CRRT) techniques have occupied an important position in the intensive care units (ICU). Serum proteome alteration and protein removal in this process are not clear. Since it has a poor understanding of mechanism of the treatment, there is a specific need of proteomics research for CRRT. The aim of this research was to study the serum proteome alterations of severe sepsis patients in the treatment of CRRT. Improved knowledge of proteome alteration could lead to the development of more efficient treatment strategies.
In this study, 20 severe sepsis patients were enrolled. A proteomic approach with two-dimensional gel electrophoresis (2-DE) coupled with mass spectrometry and bioinformatics methods was utilized to identify proteins with altered expression at different times in the treatment of continuous veno-venous haemofiltration (CVVH). All proteins were identified on the appearance of the 2-DE gel at the appropriate molecular size and pI and score from peptide mass fingerprinting. Protein identifications were confirmed by sequencing of the tryptic peptides and an independent database search based on the sequence. A further validation study was performed by western blot.
Thirty-four protein spots expressed differentially were separated. Ten proteins were identified to be the commonly differentially expressed proteins in the treatment. Seven proteins decreased in the serum and three increased.
This study gives a novel overview of serum proteome alteration of severe sepsis patient in the treatment of CVVH. Potentially interesting proteins have been revealed that are different from those identified by method of traditional biology.
连续性肾脏替代治疗（CRRT）技术在重症监护病房（ICU）中占据重要地位。在此过程中血清蛋白质组的改变以及蛋白质清除情况尚不清楚。由于对该治疗机制了解不足，因此对CRRT进行蛋白质组学研究具有特殊需求。本研究的目的是探讨连续性静脉-静脉血液滤过（CVVH）治疗中重症脓毒症患者血清蛋白质组的变化。深入了解蛋白质组的变化可能会促使开发出更有效的治疗策略。
本研究纳入了20例重症脓毒症患者。采用二维凝胶电泳（2-DE）结合质谱和生物信息学方法的蛋白质组学方法，来鉴定在CVVH治疗不同时间点表达发生改变的蛋白质。所有蛋白质均根据其在2-DE凝胶上适当的分子大小、等电点以及肽质量指纹图谱得分进行鉴定。通过胰蛋白酶肽段测序和基于序列的独立数据库搜索来确认蛋白质鉴定结果。通过蛋白质印迹法进行进一步的验证研究。
分离出34个差异表达的蛋白质斑点。鉴定出10种蛋白质为治疗中常见的差异表达蛋白质。血清中有7种蛋白质减少，3种蛋白质增加。
本研究对重症脓毒症患者在CVVH治疗中的血清蛋白质组变化给出了全新的概述。揭示了一些潜在有趣的蛋白质，它们与传统生物学方法鉴定出的蛋白质不同。
