家系早发冠心病血瘀证患者的血浆蛋白质组学分析

Analysis on plasma proteomics of the pedigree in premature coronary heart disease patients with blood stasis syndrome

目的:通过同位素相对标记与绝对定量技术(iTRAQ)分析家系早发冠心病血瘀证患者的血浆蛋白质组学变化,为今后预防疾病及临床筛查提供新的潜在生物标志物。方法:从前期流调工作中收集的冠心病患者中筛选出典型的家系早发冠心病血瘀证患者、家系早发冠心病非血瘀证患者,并匹配健康对照组,收集每位受试者的血浆样本,采用iTRAQ技术对血浆蛋白表达情况进行定量检测,对所得的蛋白数据进行鉴定与筛选,并以Western Blot验证筛选出的蛋白。结果:通过iTRAQ技术共得到差异蛋白61个,其中与健康对照组比较,家系早发冠心病血瘀证共得到32个差异蛋白,包括22个上调蛋白与10个下调蛋白;家系早发冠心病非血瘀证共得到35个差异蛋白,其中8个上调蛋白和27个下调蛋白。家系早发冠心病血瘀证组中差异蛋白的生物功能分析提示与角质化、角质化包膜及表皮结构组成部分密切联系,差异蛋白所在通路主要与金黄色葡萄球菌感染、补体系统、血小板激活等相关。与健康对照组比较,家系早发冠心病血瘀证组差异蛋白二磷酸甘油酸变位酶(BPGM)表达水平显著降低(P<0.05)。结论:经iTRAQ技术初步筛选及Western Blot验证后的差异蛋白BPGM可作为家系早发冠心病血瘀证早期诊断的潜在生物标志物之一。

Objective:To analyze the plasma proteomic changes of patients with premature coronary heart disease(PCHD)with blood stasis syndrome(BSS)by isobaric tags for relative and absolute quantification(iTRAQ),and to provide new potential biomarkers for disease prevention and clinical screening in the future.Methods:Typical familial premature coronary heart disease patients with blood stasis syndrome and familial premature coronary heart disease patients with nonblood stasis syndrome were screened from the patients with coronary heart disease in the previous epidemiological work.A healthy control group was matched to collect plasma samples from each subject.Plasma protein expression was quantitatively detected by iTRAQ technology,the obtained protein data were identified and screened,and the screened proteins were verified by Western Blot.Results:A total of 61 differential proteins were obtained through iTRAQ technology.Compared with the healthy control group,a total of 32 differential proteins were obtained in pedigree in PCHD patients with BSS,including 22 upregulated proteins and 10 down-regulated proteins.A total of 35 differential proteins were obtained in the pedigree in PCHD patients with non-BSS,including 8 up-regulated proteins and 27 down-regulated proteins.The biological function analysis of the differential protein in the pedigree in PCHD patients with BSS group indicates that it is closely related to cornification,cornified envelope and structural constituent of epidermis.The pathway of the differential protein is mainly related to staphylococcus aureus infection,complement system,platelet activation and so on.Compared with the healthy control group,the differential protein bisphosphoglycerate mutase(BPGM)expression level of the pedigree in PCHD patients with BSS group was statistically decreased(P<0.05).Conclusion:The differential protein BPGM,which has been initially screened by iTRAQ technology and verified by Western Blot,can be used as one of the potential biomarkers for the early diagnosis of the pedigree in PCHD patients with BSS.