阳虚质肠道菌群特征及其与宿主代谢共变化机制研究

Yang-deficiency constitution is commonly seen. It was mainly manifested by deficient-cold symptoms, such as fear of cold, cold limbs, etc. It's closely related to the digestive system, endocrine and metabolic diseases.Our previous study found that people with yang-deficiency constitution tend to have imbalance of intestinal flora and disorder of metabolic and immunologic function. Modern medical research has confirmed that intestinal flora involves in host metabolism and there is covariation between them. Based on these, we put forward the hypothesis of "Changes of yang-deficiency constitution's intestinal flora structure influences its metabolic function, and there exists intestinal flora with feature function intestinal flora related to metabolism". We will analyze the metabolite composition spectrum from fecal supernatant of yang-deficiency constitution based on NMR technique, and conduct identification of biological markers; We will analyze the characteristics of yang-deficiency constitution's intestinal flora based on 16S rRNA DGGE; We will correlate the DNA fingerprinting of intestinal flora structure and the NMR metabolite of metabolic changes of yang-deficiency constitution by multivariate statistical (PCA, OPLS-DA) to explore the characteristic of covariation between intestinal flora structure and host metabolism and identify the important function of intestinal flora which may regulate metabolism of yang-deficiency constitution . This research is important for establishing individualized health-care and prevention based on intestinal flora.

阳虚质是一种常见的偏颇体质，以畏寒怕冷、手足不温等虚寒表现为主要特征，与消化系统疾病、内分泌和代谢疾病的发生密切相关。前期研究发现，阳虚质存在肠道菌群失衡，及代谢、免疫功能紊乱。现代医学研究证实肠道菌群参与宿主代谢，二者有着共变化特征。据此我们提出"阳虚质肠道菌群结构的变化影响其代谢功能，并存在与代谢相关的特征功能菌群"这一假说。拟采用核磁共振技术分析阳虚质粪便上清液中的代谢物组成谱，鉴定生物标志物；通过肠道菌群的16S rRNA基因变性梯度凝胶电泳技术，分析阳虚质肠道菌群的特征；结合主成分分析、正交偏最小二乘判别分析等多变量统计学方法，将反映肠道菌群结构的DNA指纹图谱数据与反映阳虚质代谢变化的NMR代谢物组成谱数据加以关联，寻找肠道菌群结构与宿主代谢的共变化特征，鉴定具有调节阳虚质代谢的重要功能菌群。本项目对于开展基于肠道菌群的个体化诊疗与预防具有重要意义。

阳虚质是一种常见的偏颇体质，以畏寒怕冷、手足不温等虚寒表现为主要特征，与消化系统疾病、内分泌和代谢疾病的发生密切相关。本项目对阳虚质粪便样本进行NMR代谢组学分析，发现阳虚质粪便中丁酸盐、醋酸盐、蛋氨酸、琥珀酸盐、葡萄糖的浓度低于平和质；丙酸盐、丙氨酸、乳酸的浓度高于平和质组，表明阳虚质与平和质存在能量代谢、氨基酸代谢及糖代谢的差异以及相关脏腑功能的改变；采用Miseq PE300高通量测序技术对粪便样本中所有细菌16S rRNA V3-V4区进行DNA测序，发现阳虚质中短波单胞菌属（Brevundi的monas）、明串珠菌属（Leuconostoc）、Turicibacter、Defluviitaleaceae_incertae_sedis、罗氏菌属（Rothia）、Butyricimonas含量偏高；平和质中巨型球菌属（Megasphaera）、瘤胃球菌属（Ruminococcus）、毛螺菌属（Lachnospira）的含量偏高；进一步采用因子分析方法对上述数据进行相关分析，发现阳虚质代谢产物中葡萄糖含量减少，可能与明串珠菌属（Leuconostoc）、罗氏菌属（Rothia）、Defluviitaleaceae_incertae_sedis、 Butyricimonas的增多有关，与瘤胃球菌属的减少有关，其可能是影响阳虚质糖代谢、能量代谢的功能菌群。本项目揭示了阳虚质肠道菌群结构对宿主代谢的影响，为指导个体化诊疗与预防提供了依据。对于研发以肠道菌群为靶点的个性化药物与膳食干预方案、监控健康状况、有效预防相关疾病等具有重要意义。