胃癌患者苔色变化的微生态-炎症免疫-代谢机制研究

The concept of traditional Chinese medicine (TCM) tongue diagnosis holds that white coating represents the exterior syndrome and cold syndrome, yellow coating represents interior syndrome and hot syndrome, and gray and black coating always represent hot syndrome. Modern researches have proved that both cold and hot syndromes are closely related to the status of inflammation which plays an important role in the process of microflora imbalance. Based on the privious studies, this project propounds the working hypotheses that the coating-color hand in hand with the tongue-coating microflora and is the exterior image of the body's metabolism. To test the working hypotheses, the project will conduct a hospital-based case-control study with a population with 100 patients with primary gastric cancer and 120 people who have the medical exam with no severe organic wrong as control. The tongue-coating microflora is investigated by the next-generation sequencing technologies in 16S/18S rRNA genes. The serum metabonomic biomarkers of coating colors are identified by metabonomics studies on GC×GC-TOF MS. Different biomarkers associated with gene signaling pathways, such as inflammation and energy metabolism, are analyzed by network database. The inflammation signaling pathways are confirmed by quantitative analysis of 10 to 20 serum inflammation associated moleculars. The aim of this project is to draw out the assocication among gastric cancer risk, the change of coating colors and the tongue-coating microflora, serum inflammation, serum metabonomics in the patients with gastric cancer. The findings will attempts a definition of biological cause of coating color and give the scientific basis for the differentiation of TCM Zheng in gastric cancer.

中医认为，白苔主表证、寒证，黄苔主里证、热证，灰黑苔多主热证。现代研究认为寒证、热证多与机体炎症免疫状态相关，而微生态失调是炎症免疫发生的重要因素，总结既往研究工作，提出工作假说：苔色与舌苔微生态菌群结构相关，是机体炎症免疫和代谢状态的"外在映像"。本项目拟开展基于医院的病例-对照研究，原发性胃癌100例，体检无重大器质性病变的体检者120例为对照，要求性别、年龄匹配。基于微生物系统分类学的理想指标-16S/18S rRNA基因联合高通量测序，解析舌苔微生态菌群结构；基于代谢组学分析血清小分子代谢产物，鉴定胃癌患者苔色相关标志代谢分子；基于标志代谢分子网络数据库模拟炎症免疫、能量代谢等信号通路，免疫检测血清10-20个炎症分子验证机体炎症免疫途径。通过本项目研究，揭示胃癌发生、苔色变化的微生物-炎症免疫-代谢机制，阐明胃癌患者苔色变化的生物学原理，为胃癌证候规范化研究提供科学依据。

中医认为，白苔主表证、寒证，黄苔主里证、热证，灰黑苔多主热证。现代研究认为寒证、热证多与机体炎症免疫状态相关，而微生态失调是炎症免疫发生的重要因素。本项目通过基于医院的病例-对照研究，分析胃癌患者与健康对照人群舌苔微生态群落结构差异，胃癌患者不同舌苔类型微生态群落结构、免疫炎症因子网络表达及代谢组学差异比较，并联合胃癌证候和症状分析，以验证本项目提出的科研假说：苔色与舌苔微生态菌群结构相关，是机体炎症免疫和代谢状态的"外在映像。并揭示胃癌发生、苔色变化的微生物-炎症免疫-代谢机制，阐明胃癌患者苔色变化的生物学原理。研究结果显示：1）厚壁菌门/拟杆菌门比例增高、链球菌属相对丰度增加可能是影响胃癌发生的潜在危险因素。2）常见的四种舌苔类型（薄白苔、白厚苔、薄黄苔、黄厚苔）在细菌和真菌界均呈现特有的结构类型。在细菌界具有更高丰度的Lautropia属水平和更高的Capnocytophaga leadbetteri种水平，在真菌界具有更高丰度的Ampelomyces_sp_IRAN_1种水平可能是薄白苔特有微生态标记物。另外，相对于舌质，苔色与菌群结构变化的关系更加密切。菌群功能预测分析（KEGG）表明，与白苔相比，黄苔患者有15个预测功能显著增强，结合既往数据验证了黄苔患者具有更高的嘌呤代谢水平。3）较高水平的血清IL-17α水平是可能是黄苔的免疫分子标志。4）胃癌不同舌苔类型具有不同的代谢通路，其中薄白苔涉及7个特有代谢通路，白厚组涉及4个特有代谢通路，黄薄苔组涉及12个特有代谢通路，黄厚苔组涉及4个特有代谢通路。5）血清IL－1β水平可能是区分气阴两虚证与脾胃虚弱证的一个标志分子。6）相对于对照组和胃癌无症状组，血清IL-8与IL-5水平升高可能是胃癌胃脘痛患者炎症标记物。舌苔是反映胃气升降的重要外在征象，揭示胃癌患者舌苔形成的微生态-代谢-免疫标志研究机制，可以为理解胃气影响舌苔变化、胃癌证候及症状的生物学机制提供科学依据，并为研究中医药干预提供了潜在靶点和思路。