基于代谢组和转录组的慢性乙型肝炎患者不同舌象生物学基础研究

Tongue manifestation is an important appearance of our body changes, and it is also one of the core factors of TCM syndrome; therefore, revealing the biological basis of the tongue manifestation could provide a scientific evidence for the theory of TCM syndrome differentiation, which is full of importance. Our previous studies have proved the material basis of the classification of TCM syndrome differentiation can be found by systems biology methods. Thus, we propose the hypothesis that "there is a close relationship between the tongue manifestation and human body, the changes of tongue manifestation have relevant specific biological basis, and the tongue material differences and the association of its functions could be developed based on the systems biology technologies including metabonomics and transcriptomics, as well as the analysis technics of biomedical informatics grid". In this project, we will give research to chronic hepatitis B patients with red tongue with yellow coating and pale tongue with white coating, we will acquire their tongue coating and peripheral blood specimens, and get potential differences marker based on metabolomics and transcriptomics detection technologies and bioinformatics analysis techniques, then we will clear the differences in markers (spectrum) by expanding the sample verification and re-analysis. At the same time, we hope to realize the communication between the two "omic" informations through the further systems biology analysis to reveal the connection of the substance and its function, and to clarify the material foundation connection between the "outside" tongue coatings and "inherent" whole body, by which we hope to provide modern scientific basis of the tongue diagnosis and syndrome differentiation and treatment theory of TCM.

舌象是机体变化的重要窗口，是核心证素之一；揭示舌象的生物学基础，为中医临床辨证理论提供科学依据，具有重要的意义。在课题组前期研究发现系统生物学手段可揭示中医证候分类的物质基础的前提下，本课题提出 "舌象与机体状态密切联系，舌象变化有其相应的、特定的生物学基础，不同舌象可通过代谢组和转录组等系统生物学技术及生物信息网络分析技术来揭示其差异物质谱及其功能联系" 的假说。拟以慢乙肝患者舌红苔黄与舌淡苔白两种舌象为切入点，以血液和舌苔为样本，通过代谢组和转录组学两种组学技术和生物信息分析技术，获得潜在的差异标志物；通过扩大样本的验证和再分析以期明确差异标志物（谱）；同时进一步运用生物信息网络技术实现两种组学信息的沟通，揭示物质与功能的联系，揭示局部舌苔"外在"的物质基础与整体"内在"物质基础变化的联系。为中医舌诊及其辨证论治理论提供现代科学依据。

本研究以慢性乙型肝炎舌红苔黄和舌淡苔白患者为研究对象，以健康志愿者正常舌苔者为对照，共收集临床舌苔样本454例，选取典型舌象者进行舌苔样本的代谢组和16S菌群多样性分析，以及血样本转录组与代谢组的检测分析。运用生物信息分析技术进行“组学”信息的整合分析，探索物质与功能的联系。研究发现：.1慢乙肝较健康志愿者存在舌苔代谢差异，差异化合物主要为必须氨基酸类，参与缬氨酸、亮氨酸和异亮氨酸的降解以及苯丙氨酸、络氨酸代谢。慢乙肝黄苔和白苔者之间无差异化合物，但是慢乙肝黄苔患者舌苔中的氨基酸类化合物的含量均高于白苔组。.2 慢乙肝白苔、黄苔以及健康志愿者之间存在菌群差异，与慢乙肝白苔相比，黄苔患者Bacteroidetes下降，Proteobacteria 和Gracilibacteria升高，在功能预测方面，黄苔者舌苔菌OTU突出表现为参与氨基酸代谢，与代谢组结果相呼应。.3 舌苔菌群与代谢物和临床指标的关联分析提示，慢乙肝黄苔优势菌与HBV-DNA，TC等临床指标以及亮氨酸，次黄嘌呤，鸟嘌呤和甲硫氨酸等代谢物水平呈正相关，与S-腺苷-L-甲硫氨酸的水平负相关。.4 外周血转录组分析提示慢乙肝黄苔和白苔患者体内免疫状态不同，突出表现为黄苔者DC成熟通路被激活，同时伴随免疫细胞运动性增加，葡萄糖摄取增加，白苔者DC成熟通路抑制，并伴随脂肪酸和蛋白质合成的抑制。.5慢乙肝苔白者的特异性差异代谢物为酪氨酸（上调），与氨酰-tRNA生物合成有关；慢乙肝黄者脂肪类化合物如油酸、棕榈油酸、棕榈酸、月桂酸均不同程度上调，并特异性富集到脂肪酸生物合成通路。.6外周血转录组和代谢组整合分析提示苔白者潜在更低的免疫和代谢水平，而苔黄者则具有相对活跃的免疫和代谢反应，且苔黄者相对活跃的免疫和代谢反应与增加的糖酵解驱动TCA循环和PPP以促进脂肪酸合成，从而支持DC活化成熟和功能发挥有关。.本研究得到了典型慢乙肝舌红苔黄和舌淡苔白患者的特征性差异物谱，探讨外在的舌苔表象与整体内在的物质基础变化的联系。为中医舌诊及其辨证论治理论提供现代科学依据。