基于iTRAQ技术的大鼠挤压伤血清蛋白质组学分析
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摘要
目的运用蛋白质组学技术对比筛选挤压伤大鼠模型的血清差异蛋白。方法选取20只Wistar大鼠随机分为挤压伤组和对照组,每组10只。采用同位素标记相对和绝对定量技术以及液相色谱-串联质谱技术对2组的血清进行差异蛋白分析。使用Thermo Xcalibur 4.0软件采集数据,选用UniProt-Rattus norvegicus数据库进行搜索,并对差异表达的蛋白进行基因本体论富集分析以及差异蛋白通路富集分析。结果 2组血清中共有差异表达的蛋白206个,其中133个表达上调主要有14-3-3蛋白家族、M型肌酸激酶等,73个表达下调主要有小鼠球蛋白-1、血清白蛋白等。差异蛋白上调的通路共144种,下调的通路有89种。结论筛选出的差异表达蛋白可能与挤压伤的发展及预后有关,可能成为临床早期诊断及治疗的分子生物学标志物。
Objective To compare and screen serum differential proteins in rat models with crush injury by using proteomics technology. Methods A total of 20 Wistar rats were randomly divided into crush injury group and control group(n=10 rats for each group). Isotope labeling relative and absolute quantitative techniques and liquid chromatography-tandem mass spectrometry were used to analyze differential proteins in serum in two groups. Thermo Xcalibur 4.0 software was used to collect data, and UniProt-Rattus norvegicus database was used to search, and then gene ontology enrichment analysis and differential protein pathway enrichment analysis were performed for differentially expressed proteins. Results There were 206 differentially expressed proteins in two groups, among which 133 up-regulated proteins were mainly 14-3-3 protein family, M-type creatine kinase and so on, and 73 down-regulated proteins were mainly mouse globulin-1, serum albumin and so on. There are 144 up-regulated pathways and 89 down-regulated pathways for differential proteins. Conclusion Differentially expressed proteins by screening may be related to development and prognosis of crush injury, and they may be molecular biomarkers for early diagnosis and treatment.
Objective To compare and screen serum differential proteins in rat models with crush injury by using proteomics technology. Methods A total of 20 Wistar rats were randomly divided into crush injury group and control group(n=10 rats for each group). Isotope labeling relative and absolute quantitative techniques and liquid chromatography-tandem mass spectrometry were used to analyze differential proteins in serum in two groups. Thermo Xcalibur 4.0 software was used to collect data, and UniProt-Rattus norvegicus database was used to search, and then gene ontology enrichment analysis and differential protein pathway enrichment analysis were performed for differentially expressed proteins. Results There were 206 differentially expressed proteins in two groups, among which 133 up-regulated proteins were mainly 14-3-3 protein family, M-type creatine kinase and so on, and 73 down-regulated proteins were mainly mouse globulin-1, serum albumin and so on. There are 144 up-regulated pathways and 89 down-regulated pathways for differential proteins. Conclusion Differentially expressed proteins by screening may be related to development and prognosis of crush injury, and they may be molecular biomarkers for early diagnosis and treatment.
引文
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[17] 夏明,顾剑,张弛,等.血清白蛋白水平与初发急性心肌梗死的相关性研究[J].中华老年心脑血管病杂志,2018,20(2):153-157.
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[20] 杜小静,马建军,李学,等.血清白蛋白与缺血性卒中[J].国际脑血管病杂志,2015,23(7):532-536.
[21] 薛有平,雷毅,刘峰,等.短暂性脑缺血发作患者的血清生化指标的相关性研究[J].中华老年心脑血管病杂志,2018,20(3):275-277.
[2] 刘涛,白祥军.挤压伤和挤压综合征[J].创伤外科杂志,2016,18(7):447-449,封3.
[3] 潘鑫,邹圣强.挤压综合征研究进展[J].中国急救复苏与灾害医学杂志,2014,9(8):764-766.
[4] Hashemi B, Safari S, Hosseini M, et al. A systematic review of iranian experiences in seismo-nephrology[J].Arch Trauma Res, 2016,5(2):e28796.
[5] 王鑫,吕琪,侯世科.挤压伤-挤压综合征治疗的研究进展[J].中华灾害救援医学,2016,4(8):466-470.
[6] 汤欣.挤压综合征治疗研究进展[J].创伤与急危重病医学,2013,1(1): 45-48.
[7] Murata I, Nozaki R, Ooi K, et al. Nitrite reduces ischemia/reperfusion-induced muscle damage and improves survival rates in rat crush injury model[J].J Trauma Acute Care Surg, 2012,72(6):1548-1554.
[8] 顾惠珍.高压氧综合治疗四肢挤压伤的疗效观察[J].实用骨科杂志,2013,19(1):36-37.
[9] Kafle A, Puchadapirom P, Plumworasawat S, et al. Identification and characterization of protein 14-3-3 in carcinogenic liver fluke Opisthorchis viverrini[J].Parasitol Int, 2017,66(4):426-431.
[10] Lin H, Jiao X, Yu B, et al. Clinical significance of serum 14-3-3 beta in patients with hepatocellular carcinoma[J].Cancer Biomark, 2017,20(2):143-150.
[11] Ohkanda J, Kusumoto A, Punzalan L, et al. Structural effect of fusicoccin upon upregulation of 14-3-3-phospholigand interaction and cytotoxic activity[J].Chemistry, 2018,24(60):16066-16071.
[12] Rizou M, Frangou E A, Marineli F, et al. The family of 14-3-3 proteins and specifically 14-3-3σ are up-regulated during the development of renal pathologies[J].J Cell Mol Med, 2018,22(9):4139-4149.
[13] Hashemi M, Zali A, Hashemi J, et al. Down-regulation of 14-3-3 zeta sensitizes human glioblastoma cells to apoptosis induction[J].Apoptosis, 2018,23(11-12):616-625.
[14] Smani D, Sarkar S, Raymick J, et al. Downregulation of 14-3-3 proteins in a kainic acid-induced neurotoxicity model[J].Mol Neurobiol, 2018,55(1):122-129.
[15] Arques S. Serum albumin and cardiovascular diseases: a comprehensive review of the literature[J].Ann Cardiol Angeiol, 2018,67(2):82-90.
[16] 平虎,刘波.连续肾脏替代疗法治疗对挤压综合征致多器官功能障碍患者C-反应蛋白以及前白蛋白水平的影响[J].中国煤炭工业医学杂志,2014,17(6):913-915.
[17] 夏明,顾剑,张弛,等.血清白蛋白水平与初发急性心肌梗死的相关性研究[J].中华老年心脑血管病杂志,2018,20(2):153-157.
[18] Damiani E, Ince,C, Orlando F, et al. Effects of the infusion of 4% or 20% human serum albumin on the skeletal muscle microcirculation in endotoxemic rats[J].PLoS One, 2016,11(3):e0151005.
[19] Awadallah S M, Atoum M F, Nimer N A, et al. Ischemia modified albumin: an oxidative stress marker in β-thalassemia major[J].Clin Chim Acta, 2012,413(9-10):907-910.
[20] 杜小静,马建军,李学,等.血清白蛋白与缺血性卒中[J].国际脑血管病杂志,2015,23(7):532-536.
[21] 薛有平,雷毅,刘峰,等.短暂性脑缺血发作患者的血清生化指标的相关性研究[J].中华老年心脑血管病杂志,2018,20(3):275-277.