痰湿体质易感代谢性疾病的生物学基础以及调体治未病的作用机制研究

Some constitution is susceptible to specific disease.Pre-study found that risk of phlegm-dampness constitution susceptibility to metabolic disease is much higher than other constitution,and constitution adjustment could prevent the occurrence of disease.There are unique genomic characteristics (transcriptome and DNA methylation characteristics) for different constitutions including phlegm-dampness. Intestinal flora is called "second genome", co-evoluting with the host,and it can regulate the host’s gene through influencing the epigenetic processes.Pre-experiment found that phlegm-dampness constitution has a specific flora which affect the body metabolism. Therefore,w propose that the transcript expression and functional flora of phlegm-dampness constitution are the significant biological basis for its susceptibility to metabolic disease.The key point of disease occurrence and preventing is specific flora regulating the host’s specific gene / pathway through an epigenetic processe.We chose the crowd of impaired glucose tolerance,borderline elevated levels of hyperlipidemia, phlegm-dampness constitution,and balance constitution as control group, and they will be divided into four groups.For the first three group, we will implement constitution adjustment intervention .Through the study, we will find the correlation of phlegm-dampness gene, functional flora and glycolipid phenotype .Through the linkage analysis of DNA methylation level in the promoter region, we will reveal the molecular mechanism of flora regulating the host’s gene through influencing the epigenetic processese.We will further validate the findings by establishing a phlegm-dampness HFA C57BL / 6J mouse model and implementing interventions.This study will provide not only ideas for the preventing and control of metabolic diseases, but also a model for the study of disease prevention and treatment.

特定体质对特定疾病易感。前期发现痰湿质易感代谢疾病的风险远高于其他体质，调体可实现早防早治；包括痰湿质在内的不同体质类型存在独特的基因组特征（转录组和DNA甲基化特征）。而肠道菌群是与宿主协同进化的人体“第二基因组”，可通过表观基因组调控宿主基因。预实验发现痰湿质有影响机体代谢的特定菌群。据此提出，痰湿质特征性转录组表达和功能菌群是其易感代谢疾病的重要生物学基础，特定菌群通过表观途径调控宿主特定基因/通路是致病和调体治未病的关键点。选择2型糖尿病和高脂血症“病前”环节，平和质对照，对痰湿质、痰湿质糖尿病前期/血脂边缘升高者进行调体干预，鉴定痰湿特征基因、功能菌群及与糖脂表型相关性；通过启动子区DNA甲基化关联分析，揭示菌群通过表观调控宿主基因的分子机制。建立痰湿HFA C57BL/6J小鼠模型并实施干预，进一步验证上述发现。本研究不仅为代谢疾病防控提供思路，亦为调体治未病基础研究提供典范。

代谢性疾病早防早治意义重大。本项目从微生物组、代谢组、基因组等多组学入手，揭示痰湿体质易感代谢性疾病的生物学基础及调体治未病作用机制，为代谢疾病防治提供思路。.易感基础：①汇集1639项体质临床调研数据，描绘痰湿体质疾病谱；②基于242人的痰湿体质队列，发现痰湿体质量化评分与多种代谢指标具有明显的相关性；③通过临床队列“健康平和-痰湿未病-痰湿代谢综合征”三个阶段菌群和代谢组分析，联合C57BL/6J小鼠痰湿源粪菌移植实验，聚焦Howardella调控磷脂酰胆碱代谢等，阐明痰湿体质易感代谢紊乱的肠道微生物机制。④通过临床队列lcnRNA-mRNA、miRNA表达谱检测，聚焦炎症反应、免疫应答和糖脂代谢通路，阐明痰湿体质易感代谢紊乱的基因组基础。.调体机制：①通过249例随机、双盲、安慰剂对照的大样本临床试验，证实化痰祛湿方能够实现糖脂代谢性疾病的早期联合防治；②通过化痰祛湿方干预及菌群移植等多个动物实验，夯实调体防治代谢性疾病的菌群机制；③聚焦痰湿血脂异常人群，开展差异菌种、循环代谢物和DNA甲基化相关性研究，筛选Dmr4179等甲基化位点及L-Acetylcarnitine等差异代谢物，探索了化痰祛湿方通过“菌群-代谢物-DNA甲基化”轴改善痰湿体质、减轻血脂异常状态的可能机制。④基于临床动态观察及高通量miRNA测序、PT-qPCR验证，证实化痰祛湿方可通过调节hsa-miR-1237-3p作用于CRK和HGF，并影响下游PI3K/AKT通路，逆转痰湿体质代谢紊乱。.药效物质基础：采用UPLC鉴定了化痰祛湿方中的小分子和多糖类大分子物质，进一步联合液质联用、网络药理、分子对接和动物实验，揭示化痰祛湿方中芦丁等9个化合物通过对接IGF1R开启PI3K/Akt通路改善脂代谢；结合16sRNA基因测序及靶向代谢组检测，认为降低Fam.丰度改善IR等可能是化痰祛湿方糖类物质调节菌群改善代谢的关键机制。