基于iTRAQ技术的家系早发冠心病血瘀证外周血差异蛋白表达分析  

Differential protein expression in peripheral blood of Pedigree in premature coronary heart disease with blood stasis syndrome based on iTRAQ technology

作　　者：张梦雪 李仙福 张书萌 于子璇 刘佳[1] 江雨洁 谢婷 陈伶利[4] 李杰[1] ZHANG Meng-xue;LI Xian-fu;ZHANG Shu-meng;YU Zi-xuan;LIU Jia;JIANG Yu-jie;XIE Ting;CHEN Ling-li;LI Jie(Hunan University of Chinese Medicine,Changsha 410208,China;Longhua Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China;Ganzhou Hospital of Traditional Chinese Medicine,Ganzhou 341000,China;Hunan Provincial Key Laboratory of Pathogenic Biology Based on Integrated Chinese and Western Medicine,Hunan University of Chinese Medicine,Changsha 410208,China)

机构地区：[1]湖南中医药大学,长沙410208 [2]上海中医药大学附属龙华医院,上海200032 [3]赣州市中医院,赣州341000 [4]湖南中医药大学中西医结合病原生物学湖南省重点实验室,长沙410208

出　　处：《中华中医药杂志》2023年第3期1196-1201,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81874375);湖南省自然科学基金项目(No.2020JJ4464);湖南省中医药科研计划重点项目(No.2021024);湖南中医药大学校级科研重点项目(No.2020XJJJ002)。

摘　　要：目的:应用同位素标记相对和绝对定量(iTRAQ)技术研究家系早发冠心病(PCHD)血瘀证患者外周血浆差异蛋白的表达情况,为研究疾病的发生机制提供依据。方法:2019年8月至12月在湖南中医药大学第一附属医院和长沙金润中医院收集冠心病病例101例,从中筛选出家系PCHD血瘀证和家系PCHD非血瘀证患者各3例,并匹配健康对照组2例。采用iTRAQ技术对两组进行蛋白质组学研究,筛选出家系PCHD血瘀证的血浆差异蛋白。采用Western Blot验证差异蛋白纤维蛋白原γ链(FGG)、纤维蛋白原β链(FGB)。结果:与健康对照组相比,家系PCHD血瘀证组有差异表达蛋白32个(上调蛋白22个,下调蛋白10个),对差异蛋白进行基因本体(GO)富集和京都基因与基因组百科全书(KEGG)通路分析,发现其生物学过程主要与角质细胞分化、纤维蛋白溶解、表皮结构组成等相关;其相关通路主要涉及金黄色葡萄球菌感染、补体系统、血小板激活等通路,其中FGG、FGB共同参与通路补体系统、血小板激活。对差异蛋白进行相互作用分析,发现多个蛋白存在相互联系,其中以FGG、FGB相互作用关系显著。验证结果显示,与健康对照组比较,家系PCHD血瘀证组FGG、FGB蛋白表达量显著增加(P<0.01)。结论:FGB、FGG蛋白表达上调可能是家系PCHD血瘀证发病机制之一。Objective:To study the expression of peripheral plasma differential protein in patients with blood stasis syndrome(BBS)of pedigree in premature coronary heart disease(PCHD)with isotope labeling relative and absolute quantitative(iTRAQ)technology,and to provide evidence for studying the mechanism of disease development.Methods:From August to December 2019,101 cases of CHD were collected at the First Affiliated Hospital of Hunan University of Chinese Medicine and Changsha Jinrun Hospital of Traditional Chinese Medicine,from which 3 patients each with pedigree in PCHD with BSS and pedigree in PCHD with non-BSS were screened,and 2 healthy controls were matched.The iTRAQ technology was used to perform proteomic study on each groups to screen out the plasma differential proteins of pedigree in PCHD with BSS.Western Blot was used to verify the differential protein fibrinogenγchain(FGG)and fibrinogenβchain(FGB).Results:Compared with the healthy control group,there were 32 differentially expressed proteins(22 up-regulated proteins and 10 down-regulated proteins)in the pedigree PCHD with BSS group.Gene ontology(GO)enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis of the differential protein revealed that its biological processes are mainly related to keratinocyte differentiation,fibrinolysis,and epidermal structure composition;its related pathways mainly involve Staphylococcus aureus infection,complement system,platelet activation and other pathways,in which FGG and FGB are jointly involved in the pathway complement system and platelet activation.Interaction analysis of the differential proteins revealed that several proteins were interconnected,among which FGG and FGB were significantly related to each other.The validation results showed that the protein expressions of FGG and FGB were significantly increased in the pedigree PCHD with BSS group compared with the healthy control group(P<0.01).Conclusion:Up-regulation of FGB and FGG protein expression may be one of the pathogenesis of

关 键 词：早发冠心病 血瘀证 家系 ITRAQ 纤维蛋白原γ链 纤维蛋白原β链 

分 类 号：R259[医药卫生—中西医结合]