基于湿热型与寒湿型泄泻病证探究舌面与肠道菌群的生物学特征

Exploration on the biological characteristics of tongue and intestinal flora based on damp-heat and cold-damp diarrhea

目的:通过检测与分析湿热型、寒湿型泄泻患者及健康人群舌面与肠道菌群特征,阐释中医“舌为脾之外候”的生物学基础。方法:采集湿热型、寒湿型泄泻患者及健康志愿者舌苔、粪便样本各12例,提取DNA,采用IlluminaMiseq测序平台进行16SrRNA高通量测序,获得样本物种信息,对各组间舌、肠菌群的多样性、物种差异性进行分析。结果:受试者舌、肠菌群丰富度改变不明显,但菌群结构改变明显,3组样本的舌、肠菌群各自聚类;差异分析发现,泄泻患者舌、肠菌群的变化趋势具有一致性,均表现为拟杆菌丰度降低,湿热型低于寒湿型;变形菌、放线菌丰度升高,湿热型高于寒湿型,而厚壁菌门在寒湿型中最高,正常组次之。结论:变形菌、放线菌丰度的增加与泄泻的发生相关,而厚壁菌、拟杆菌是区别湿热型泄泻与寒湿性泄泻的主要菌种,舌、肠菌群的一致性变化可作为阐述中医“舌为脾之外候”理论的生物学基础。

Objective: Through the detection and analysis of the characteristics of the tongue and intestinal flora in patients with damp-heat and cold-damp diarrhea and healthy people, to explain the biological basis of ’tongue as a symptom outside of spleen of TCM’. Methods: Each of 12 samples of tongue coating and feces from patients with damp-heat type and colddamp type diarrhea and healthy volunteers were collected to extract DNA, and Illumina Miseq sequencing platform for 16 S rRNA high-throughput sequencing was used to obtain the biological species information of the samples. The diversity, species differences of tongue and intestinal flora among each group were analyzed. Results: The richness of tongue and intestine flora of the subject’s did not change significantly, but the structure of the flora was significantly different. The tongue and intestinal flora of the three groups of samples were clustered separately. The difference analysis found that the changes in the tongue and intestinal flora of diarrhea patients were consistent, all showed that the abundance of Bacteroides decreased, the damp-heat type was lower than that of the cold-damp type;the abundance of Proteobacteria and actinomycetes increased, and the damp-heat type was higher than that of the cold-damp type. Firmicutes was the highest in the cold-damp type, the normal group came next. Conclusion: The increase in the abundance of proteobacteria and actinomycetes is related to the occurrence of diarrhea, while firmicutes and bacteroides are the main bacterial species that distinguish between damp-heat diarrhea and cold-damp diarrhea. The consistent changes of the tongue and intestinal flora can be used as the biological basis for explaining the theory of ’the tongue as a symptom outside of the spleen’ in TCM.