Differences in serum proteins in traditional Chinese medicine constitutional population: analysis and verification

Using high-throughput antibody microarray, through cross-sectional sample detection and verification of samples that had undergone physical changes over time, it was found that people with balanced constitution and dampness constitution in Chinese medicine showed differences in serum protein expression. The differences were expressed as the level changes of 19 proteins such as Dectin-2, Siglec-7, AIF and TACI. The research results provided the basis for the scientific expression of traditional Chinese medicine (TCM) constitution. We prospectively enrolled two cohorts into for our study. The first cohort included 4 balanced constitution (BC) individuals transformed to dampness constitution (DC), another 3 DC individuals transformed to BC. The second cohort contained 38 DC individuals and 37 BC individuals with no diagnosed diseases. Whole blood was collected in vacutainer serum tubes, and then 1000 serum proteins were analyzed simultaneously using an antibody-based array. We identified and verified 19 different serum proteins by high-throughput antibody microarray technology in BC and DC group, as well as serum profiles with constitution changes at different time points. Meanwhile, these differential proteins were closely related to clinical information such as human lifestyle and characteristics by established SVM model, sensitivity and specificity screening. We found that the main functions of these differential proteins were mainly focused on the immunity and inflammation by functional analysis, which might be the biological basis for the differences between BC and DC group.

本研究采用高通量抗体微阵列（high-throughput antibody microarray）技术，通过对随时间发生体质变化的样本开展横断面检测与验证，发现中医平和体质与痰湿体质人群的血清蛋白表达存在差异，该差异表现为Dectin-2、Siglec-7、AIF及TACI等19种蛋白的水平变化。本研究结果为中医体质的科学化表达提供了依据。本研究前瞻性招募两队列人群参与研究：第一队列包含4例由平和体质（balanced constitution, BC）转化为痰湿体质（dampness constitution, DC）的受试者，以及3例由DC转化为BC的受试者；第二队列纳入38例DC受试者与37例无确诊疾病的BC受试者。研究采用真空血清采血管采集全血，随后基于抗体芯片同步分析1000种血清蛋白。本研究借助高通量抗体微阵列技术，在BC组与DC组以及不同时间点发生体质变化的受试者血清中，鉴定并验证了19种差异血清蛋白，同时获得了对应血清表达谱。通过构建支持向量机（Support Vector Machine, SVM）模型并结合灵敏度与特异性筛选，发现上述差异蛋白与受试者的生活方式、特征等临床信息密切相关。经功能分析发现，上述差异蛋白的主要功能集中于免疫与炎症过程，这可能是BC组与DC组人群体质差异的生物学基础。