基于iTRAQ技术筛选阴虚“上火”人群血清生物标志物
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摘要
目的:筛选阴虚上火人群血清差异表达蛋白,探讨阴虚上火的现代生物学基础,为上火机制的深入研究提供科学依据。方法:应用iTRAQ技术检测血清蛋白质,结合GO、KEGG及STRING等方法筛选血清差异蛋白,采用ELISA进行验证。结果:载脂蛋白C3(APOC3)、锌α2糖蛋白(ZA2G)和C-反应蛋白(CRP)等14个蛋白显著上调(P<0.05),而凝溶胶蛋白(GELS)、凝血酶原(THRB)、抗凝血酶-Ⅲ(ANT3)及血小板糖蛋白V(GPV)等25个蛋白显著下调(P<0.05)。APOC3、ZA2G和GELS蛋白变化与iTRAQ鉴定结果相符。结论:血清APOC3、ZA2G和GELS可作为阴虚上火人群潜在的生物标志物。
Objective: To screen and identify serum potential biomarkers in yin deficiency 'Shanghuo' patients, discuss the modern biological basis of yin deficiency 'Shanghuo' and provide scientific references for further study of the mechanism of'Shanghuo'. Methods: The serum proteins were detected by isobaric tags for relative and absolute quantification(iTRAQ). The differential proteins were selected by bioinformatics analysis methods such as GO, KEGG and STRING. Sample validation were conducted by enzyme-linked immunosorbent assay(ELISA). Results: Forteen proteins including APOC3, ZA2 G and CRP were significantly up-regulated(P<0.05), and 25 proteins including, GELS, THRB, ANT3 and GPV were significantly down-regulated(P<0.05). APOC3, ZA2 G and GELS protein expression validated by ELISA were consistent with iTRAQ results. Conclusion: Serum APOC3, ZA2 G and GELS can be used as the potential biomarkers for the yin deficiency 'Shanghuo'.
Objective: To screen and identify serum potential biomarkers in yin deficiency 'Shanghuo' patients, discuss the modern biological basis of yin deficiency 'Shanghuo' and provide scientific references for further study of the mechanism of'Shanghuo'. Methods: The serum proteins were detected by isobaric tags for relative and absolute quantification(iTRAQ). The differential proteins were selected by bioinformatics analysis methods such as GO, KEGG and STRING. Sample validation were conducted by enzyme-linked immunosorbent assay(ELISA). Results: Forteen proteins including APOC3, ZA2 G and CRP were significantly up-regulated(P<0.05), and 25 proteins including, GELS, THRB, ANT3 and GPV were significantly down-regulated(P<0.05). APOC3, ZA2 G and GELS protein expression validated by ELISA were consistent with iTRAQ results. Conclusion: Serum APOC3, ZA2 G and GELS can be used as the potential biomarkers for the yin deficiency 'Shanghuo'.
引文
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[12]Jin G Z,Li Y,Cong W M,et al.ITRAQ-2DLC-ESI-MS/MS based identification of a new set of immunohistochemical biomarkers for classification of dysplastic nodules and small hepatocellular carcinoma.J Proteome Res,2011,10(8):3418-3428
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[14]Larsson M,Vorrsjo E,Talmud P,et al.Apolipoproteins C-Ⅰand C-Ⅲinhibit lipoprotein lipase activity by displacement of the enzyme from lipid droplets.J Biol Chem,2013,288(47):33997-34008
[15]Wu J J,Rowan C G,Bebchuk J D,et al.Association between tumor necrosis factor inhibitor(TNFi)therapy and changes in C-reactive protein(CRP),blood pressure,and alanine aminotransferase(ALT)among patients with psoriasis,psoriatic arthritis,orrheumatoid arthritis.J Am Acad Dermatol,2015,72(5):917-919
[16]唐茜茜,王颖,王寿勇.凝溶胶蛋白与炎症反应的关系.医学综述,2017,23(2):231-234
[17]Zhu Y,Qin Z,Gao J,et al.Vitamin D therapy in experimental allergic encephalomyelitis could limited by opposing effects of sphingosine 1-phosphate and gelsolin dysregulation.Mol Neurobiol,2014,50(3):733-743
[18]Kristin Eckardt,Annette Schober,Birgit Platzbecker,et al.The adipokine zinc-α2-glycoprotein activates AMP kinase in human primary skeletal muscle cells.Arch Physiol Biochem,2011,117(2):88-93
[19]Tedeschi S,Pilotti E,Parenti E,et al.Serum adipokine zinca2-glycoprotein and lipolysis in cachectic and noncachectic heart failure patients:Relationship with neurohormonal and inflammatory biomarkers.Metabolism,2012,61(1):37-42
[2]余凯,钱俊华,范永升.以离位之相火论“上火”之火.中华中医药杂志,2017,32(8):3365-3367
[3]唐晓阳,谢冠群,范永升.现代医学对上火的认识及其对中医治疗的启示.浙江临床医学,2013,15(3):389-390
[4]于洪洁,王勇,杨丽萍.阳虚、阴虚体质在大学生中的流行病学调查.辽宁中医杂志,2013,40(4):619-621
[5]Gianazza E,Miller I,Palazzolo L,et al.With or without you-Proteomics with or without major plasma/serum proteins.J Proteomics,2016,140:62-80
[6]王秋红,王悦,杨欣,等.基于i TRAQ技术的血清蛋白质组学研究进展.辽宁中医药大学学报,2017,1(1):5-7
[7]Anderson N L,Matheson A D,Steiner S.Proteomics:Applications in basic and applied biology.Curr Opin Biotechnol,2000,11(4):408-412
[8]周佳,吴德鸿,韦双双,等.基于GC-MS对中医实热“上火”血清代谢特征的研究.中华中医药杂志,2017,32(4):1789-1792
[9]中华中医药学会.中医体质分类与判定.北京:中国中医药出版社,2009
[10]Cao W,Zhou Y,Li Y,et al.ITRAQ-based proteomic analysis of combination therapy with taurine,epigallocatechin gallate,and genistein on carbon tetrachloride-induced liver fibrosis in rats.Toxicol Lett,2015,232(1):233-245
[11]Wang Y S,Cao R,Jin H,et al.Altered protein expression inserum from endometrial hyperplasia and carcinoma patients.J Hematol and Oncol,2011,4:15
[12]Jin G Z,Li Y,Cong W M,et al.ITRAQ-2DLC-ESI-MS/MS based identification of a new set of immunohistochemical biomarkers for classification of dysplastic nodules and small hepatocellular carcinoma.J Proteome Res,2011,10(8):3418-3428
[13]Kang Su H,Kim Y R,Hong S H.Synergic effects of the ApoC3and ApoA4 polymorphisms on the risk of hypertension.Genes Genom,2017,39:1163-1172
[14]Larsson M,Vorrsjo E,Talmud P,et al.Apolipoproteins C-Ⅰand C-Ⅲinhibit lipoprotein lipase activity by displacement of the enzyme from lipid droplets.J Biol Chem,2013,288(47):33997-34008
[15]Wu J J,Rowan C G,Bebchuk J D,et al.Association between tumor necrosis factor inhibitor(TNFi)therapy and changes in C-reactive protein(CRP),blood pressure,and alanine aminotransferase(ALT)among patients with psoriasis,psoriatic arthritis,orrheumatoid arthritis.J Am Acad Dermatol,2015,72(5):917-919
[16]唐茜茜,王颖,王寿勇.凝溶胶蛋白与炎症反应的关系.医学综述,2017,23(2):231-234
[17]Zhu Y,Qin Z,Gao J,et al.Vitamin D therapy in experimental allergic encephalomyelitis could limited by opposing effects of sphingosine 1-phosphate and gelsolin dysregulation.Mol Neurobiol,2014,50(3):733-743
[18]Kristin Eckardt,Annette Schober,Birgit Platzbecker,et al.The adipokine zinc-α2-glycoprotein activates AMP kinase in human primary skeletal muscle cells.Arch Physiol Biochem,2011,117(2):88-93
[19]Tedeschi S,Pilotti E,Parenti E,et al.Serum adipokine zinca2-glycoprotein and lipolysis in cachectic and noncachectic heart failure patients:Relationship with neurohormonal and inflammatory biomarkers.Metabolism,2012,61(1):37-42