基于iTRAQ标记结合生物信息学技术的风湿性疾病湿热证血清蛋白组学研究
Study on Serum Proteomics of Rheumatism with Damp-heat Syndrome Based on iTRAQ Labeling Method Combined with Bioinformatics Technology
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摘要: 目的 应用iTRAQ标记结合生物信息学分析技术,开展风湿性疾病湿热证的血清蛋白组学研究。方法 对强直性脊柱炎湿热证、痛风湿热证、正常对照共3组各20例血清,进行iTRAQ标记结合液相串联质谱蛋白组学分析,应用DAVID、UniProtKB/Swiss-Prot、IPA等网络数据库平台对风湿性疾病湿热证的差异蛋白进行生物信息学分析,并对显著差异蛋白进行验证。结果 获得15个湿热证差异蛋白,其中表达上调的蛋白12种,下调蛋白3种。通过分析得到差异表达蛋白所涉及的5个最有意义的典型生物信号通路。这些差异蛋白功能及生物信息学分析提示与疾病急性期反应和疾病活动度有关。验证实验结果与iTRAQ检测结果一致。结论 为风湿性疾病湿热证的分子机制以及候选诊断治疗标志物的进一步研究提供了依据。Abstract: OBJECTIVE To investigate the serum proteomics of rheumatism with damp-heat syndrome by using iTRAQ labeling method together with bioinformatics technology. METHODS 20 serum samples respectively from ankylosing spondylitis with damp-heat syndrome group, arthrolithiasis with damp-heat syndrome group and normal control group were collected for proteomics analysis through iTRAQ labeling and liquid chromatography. Such network database platforms as DAVID, UniProtKB/Swiss Prot and IPA were resorted to perform bioinformatics analysis of differential protein in rheumatic diseases with damp-heat syndromes and verification would be done to the significant differential proteins. RESULTS Fifteen differential proteins of dampness-heat syndrome were obtained, with twelve being the up-regulated expression proteins and three being the down-regulated expression proteins. The five typical biological signaling pathways of most significant meaning related to differential protein expression were obtained through analysis. These differential protein functions and bioinformatic analysis were proved to be related to the acute phase responses and activities of diseases. Verification test results were consistent with those of iTRAQ. CONCLUSION This provides evidences for further study on molecular mechanism of rheumatic diseases with damp-heat syndromes as well as the candidate diagnostic markers.
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Key words:
- ankylosing spondylitis /
- gout /
- damp-heat syndrome /
- serum /
- proteomics
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[1] 张英泽,阎小萍,赵铁.风湿病湿热证的实质研究辨析[J].中医研究,2011, 24(11):1-3. [2] ZHANG YZ, YAN XP, ZHAO T. Substantive research discrimination of rheumatic diseases with heat syndrome[J]. Tradit Chin Med Res, 2011, 24(11): 1-3. [3] 赵铁,殷婷婷,张英泽,等.基于代谢组学的强直性脊柱炎和痛风性关节炎湿热证共性特征研究[J].中医杂志,2013, 54(7):592-596. [4] ZHAO T, YAN TT, ZHANG YZ, et al. Study on common features of ankylosing spondylitis and damp-heat syndrome in gouty arthritis based on metabolomics[J]. J Tradit Chin Med, 2013, 54(7):592-596. [5] 殷婷婷, 赵春杰, 张英泽, 等. 风湿性疾病湿热证实质的脂质组学研究[J]. 中华中医药杂志, 2013, 28(1): 210-3. [6] YIN TT, ZHAO CJ, ZHANG YZ, et al. Lipid group study on the substantive of rheumatic diseases with damp-heat syndrome[J]. Chin J Tradit Chin Med Pharm, 2013, 28(1) : 210-213. [7] 赵铁,张英泽,王晓雪,等.基于微量元素组学的风湿性疾病湿热证共性特征研究[J].中医杂志,2014, 55(24):2100-2105. [8] ZHAO T, ZHANG YZ, WANG XX, et al. Study on the common features in damp-heat syndrome of rheumatic diseases based on Trace element profiling[J]. J Tradit Chin Med, 2014, 55(24) : 2100-2105. [9] VAN DER LINDEN S, VALKENBURG HA, CATS A. Evaluation of diagnostic criteria for ankylosing spondylitis. A proposal for modification of the New York criteria[J]. Arthritis Rheum, 1984, 27(4): 361-368. [10] WALLACE SL, ROBINSON H, MASI AT, et al. Preliminary criteria for the classification of the acute arthritis of primary gout[J]. Arthritis Rheum, 1977, 20(3): 895-900. [11] 中药新药临床研究指导原则[M].北京:中国医药科技出版社,2002. [12] Guiding Principles of Clinical Research on Chinese Traditional Medicine New Drug[M]. Beijing: Chinese medical science and technology press, 2002. [13] BENHAMOU M, GOSSEC L, DOUGADOS M. Clinical relevance of C-reactive protein in ankylosing spondylitis and evaluation of the NSAIDs/coxibs' treatment effect on C-reactive protein[J]. Rheumatology (Oxford), 2010, 49(3): 536-541. [14] JUNG SY, PARK MC, PARK YB, et al. Serum amyloid a as a useful indicator of disease activity in patients with ankylosing spondylitis[J]. Yonsei Med J, 2007, 48(2): 218-224. [15] WATSON CT, BREDEN F. The immunoglobulin heavy chain locus: genetic variation, missing data, and implications for human disease[J]. Genes Immun, 2012, 13(5): 363-373. [16] GOTTENBERG JE, AUCOUTURIER F, GOETZ J, et al. Serum immunoglobulin free light chain assessment in rheumatoid arthritis and primary Sjogren's syndrome[J]. Ann Rheum Dis, 2007, 66(1): 23-27. [17] DING PH, JIN LJ. The role of lipopolysaccharide-binding protein in innate immunity: a revisit and its relevance to oral/periodontal health[J]. J Periodontal Res, 2014, 49(1): 1-9. [18] LONDONO J, ROMERO-SANCHEZ MC, TORRES VG, et al. The association between serum levels of potential biomarkers with the presence of factors related to the clinical activity and poor prognosis in spondyloarthritis[J]. Rev Bras Reumatol, 2012, 52(4): 536-544. [19] CESARO A, ANCERIZ N, PLANTE A, et al. An inflammation loop orchestrated by S100A9 and calprotectin is critical for development of arthritis[J]. PLoS One, 2012, 7(9): e45478. [20] KALSHEKER NA. Alpha 1-antichymotrypsin[J]. Int J Biochem Cell Biol, 1996, 28(9): 961-964. [21] CHARD MD, CALVIN J, PRICE CP, et al. Serum alpha 1 antichymotrypsin concentration as a marker of disease activity in rheumatoid arthritis[J]. Ann Rheum Dis, 1988, 47(8): 665-671. [22] GOICOECHEA DJE, CAESAR JJ, MALIK TH, et al. Dimerization of complement factor H-related proteins modulates complement activation in vivo[J]. Proc Natl Acad Sci USA, 2013, 110(12): 4685-4690. [23] SUN S, HEMRIKSEN K, KARSDAL MA, et al. Measurement of a MMP-2 degraded Titin fragment in serum reflects changes in muscle turnover induced by atrophy[J]. Exp Gerontol, 2014, 58(1): 83-89. [24] WOUTERS D, VAN SCHOUWENBURG P, VAN DER HORST A, et al. High-throughput analysis of the C4 polymorphism by a combination of MLPA and isotype-specific ELISA's[J]. Mol Immunol, 2009, 46(4): 592-600. [25] OLSSON B, GIGANTE B, MEHLIG K, et al. Apolipoprotein C-I genotype and serum levels of triglycerides, C-reactive protein and coronary heart disease[J]. Metabolism, 2010, 59(12): 1736-1741. [26] GENRE F, LOPEZ-MEJIAS R, MIRANDA-FILLOY JA, et al. Gelsolin levels are decreased in ankylosing spondylitis patients undergoing anti-TNF-alpha therapy[J]. Clin Exp Rheumatol, 2014, 32(2): 218-224. [27] TANG H, MIRSHAHIDI S, SENTHIL M, et al. Down-regulation of LXR/RXR activation and negative acute phase response pathways in colon adenocarcinoma revealed by proteomics and bioinformatics analysis[J]. Cancer Biomark, 2014, 14(5): 313-324. [28] VAVASSORI P, MENCARELLI A, RENGA B, et al. The bile acid receptor FXR is a modulator of intestinal innate immunity[J]. J Immunol, 2009, 183(10): 6251-6261. [29] RITTER SY, SUBBAIAH R, BEBEK G, et al. Proteomic analysis of synovial fluid from the osteoarthritic knee: comparison with transcriptome analyses of joint tissues[J]. Arthritis Rheum, 2013, 65(4): 981-992.
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