冠心病痰浊证与健康人血浆外泌体蛋白质谱研究

A Study of Plasma Exosome Protein Spectrum in Coronary Heart Disease Patients with Phlegm Turbidity Syndrome and Healthy People

目的探索冠心病痰浊证患者血浆外泌体中特异蛋白表达,为冠心病痰浊证的诊断提供生物学参考。方法纳入冠心病痰浊证患者20例和同时期健康体检者20例,运用qEV尺寸排阻法提取所有受试者血浆外泌体,随机抽取10例冠心病痰浊证和10例健康受试者,运用label free非靶向蛋白定量分析,筛选两组间差异表达蛋白,并运用MRM靶向蛋白标记技术在剩余受试者中验证差异表达蛋白。结果与健康对照组相比,冠心病痰浊证患者血浆外泌体差异表达蛋白共计214个(61个上调,153个下调),主要与胆固醇代谢、补体和凝血级联、免疫效应有关。ANXA6、C4BPB、F8、CFB、APOE、C9、CLU蛋白进一步通过了MRM靶向蛋白验证。结论冠心病痰浊证患者血浆外泌体蛋白表达与健康人存在差异,差异蛋白主要与胆固醇代谢、补体和凝血级联有关。

Objective To explore the specific plasma exosomal protein expression in Coronary Heart Disease(CHD)with phlegm-turbidity syndrome(PTS),so as to provide biological reference for the diagnosis of CHD with PTS.Methods A total of 20 CHD patients with PTS and 20 healthy subjects at the same period were included.Plasma exosomes were extracted from all subjects using the qEV size exclusion method.First,10 CHD cases with PTS and 10 healthy subjects were randomly selected,and differentially expressed proteins between the two groups were screened using label free non-targeted protein quantification analysis.Then,MRM targeting protein labeling technique was applied to verify the differentially expressed proteins in the remaining subjects.Results Compared with healthy controls,214 differentially expressed plasma exosomal proteins(61 up-regulated and 153 down-regulated)were found in CHD patients with PTS,mainly related to cholesterol metabolism,complement and coagulation cascade,and immune effects.ANXA6,C4BPB,F8,CFB,APOE,C9,and CLU proteins were further validated by MRM targeting protein.Conclusion CHD patients with PTS had differences in plasma exosomal protein expression from healthy controls,and the differential proteins are mainly related to cholesterol metabolism,complement,and the coagulation cascade.