非酒精性脂肪肝湿热蕴结证肠道菌群特征及其与宿主代谢共变化机制研究

Non-alcoholic fatty liver disease has become a common and frequently occurring disease. Recent research showed that gut microbiota played key roles in the development of non-alcoholic fatty liver disease, and accumulation of damp heat syndrome as its common syndrome had imbalance of gut microbiota and metabolic disorders. Research showed that gut microbiota affected host metabolism, and there was co-variation between them, and there existed gut microbiota which were associated with metabolism. Therefore we put forward the hypothesis of “imbalance of gut microbiota in non-alcoholic fatty liver disease with accumulation of damp heat syndrome affect its metabolic function, and there exists gut microbiota which are associated with metabolism”. This project is conducting non-alcoholic fatty liver disease with accumulation of damp heat syndrome patients as the research object, using gut microbiota and co-metabolism between gut microbiota and host as the starting point, taking advantages of 16S rRNA amplication sequencing technique, GC/TOFMS technique and multivariate statistical analysis to explore gut microbiota and urine and fecal metabolites in non-alcoholic fatty liver disease with accumulation of damp heat syndrome, and to find the characteristics of co-variation between gut microbiota and host metabolism and to identify gut microbiota which are associated with metabolism. It helps to objectify accumulation of damp heat syndrome of non-alcoholic fatty liver and promote the development of individual diagnosis and treatment based on gut microbiota.

非酒精性脂肪肝为临床常见病、多发病。近期研究表明肠道菌群在非酒精性脂肪肝发生发展中起到关键作用，其常见证候湿热蕴结证存在肠道菌群失衡和代谢功能紊乱。文献报道肠道菌群影响宿主代谢，二者有着共同变化的特征，同时存在代谢相关的功能菌群。因此我们提出：“非酒精性脂肪肝湿热蕴结证肠道菌群失衡影响其代谢功能，同时存在代谢相关的功能菌群”假说。课题组以非酒精性脂肪肝湿热蕴结证患者为研究对象，以肠道菌群及其与宿主代谢共变化为切入点，利用16S rRNA扩增子测序技术、GC/TOFMS技术和多变量统计分析方法，探讨非酒精性脂肪肝湿热蕴结证的肠道菌群特征和尿液、粪便代谢物特征，阐明肠道菌群与宿主代谢的共变化特征，揭示肠道菌群对其代谢功能影响，确定代谢相关的功能菌群。本研究有助于非酒精性脂肪肝湿热蕴结证客观化，对于开展基于肠道菌群的个体化诊疗具有重要意义。

肠道菌群在NAFLD发生发展中起到关键作用，其常见证候湿热蕴结证存在肠道菌群失衡和代谢功能紊乱。本课题采用16SrRNA 扩增子测序技术和代谢组学技术，深入探究NAFLD湿热蕴结证肠道菌群特征和代谢物的特征，揭示肠道菌群对宿主代谢的影响。粪便代谢组数据显示：与健康组和NAFLD肝郁脾虚组比，NAFLD湿热蕴结组苯丙氨酸、氨、PA(15:0/24:0)、嘌呤水平均显著升高。与健康组比，甘氨鹅去氧胆酸、脱氧胆酸、胆酸、尿酸、乙醛、黄尿酸、色胺、犬尿氨酸水平均显著升高，维生素C、4-羟苯丙酮酸、胆碱、1-磷酸鞘氨醇、醋酸盐、色氨酸、甘氨胆酸、α亚麻酸水平均显著降低。血清代谢组数据显示：与健康组和NAFLD肝郁脾虚组比，NAFLD湿热蕴结组色氨酸、Inosine 5'-Monophosphate、鸟嘌呤核苷水平均显著升高，甲状腺素水平显著降低。与健康组比，NAFLD湿热蕴结组苯丙氨酸水平显著升高，与NAFLD肝郁脾虚组比，苯丙氨酸水平显著下降。与健康组比，NAFLD湿热蕴结组吲哚乙腈水平显著降低，甘氨鹅去氧胆酸、甘氨胆酸盐、TG水平均显著升高。尿液代谢组数据显示：与健康组比，NAFLD湿热蕴结组TG、Uric acid、FA 18:0、PE、Pyruvic acid、2-Hydroxycinnamic acid水平显著提高；PC水平显著下降。NAFLD湿热蕴结证与志贺氏菌属、柯林斯氏菌属、土壤杆菌属、双歧杆菌属、Butyricimonas、Eggerthella、Subdoligranulum、Stenotrophomonas、假单胞菌属、不粘柄菌属、巨单胞菌属、不动杆菌属、明串珠菌属、毛螺菌属、Odoribacter、Anaerofustis等菌群失调密切相关。NAFLD湿热蕴结证肠道菌群失调与宿主色氨酸-犬尿氨酸代谢紊乱、苯丙氨酸-酪氨酸代谢紊乱、胆汁酸代谢紊乱、嘌呤代谢紊乱、胆碱代谢紊乱、α亚麻酸代谢紊乱、1-磷酸鞘氨醇代谢紊乱、维生素C代谢紊乱、赖氨酸代谢紊乱密切相关。NAFLD湿热蕴结证存在肠道菌群和代谢紊乱，肠道菌群可能通过调控色氨酸代谢、苯丙氨酸代谢、胆汁酸代谢、嘌呤代谢、维生素C、磷脂代谢、短链脂肪酸等途径参与NAFLD湿热蕴结证的发生发展。有助于从新的视角阐明NAFLD湿热蕴结证的病机，并为NAFLD的中医药诊疗提供新思路和新靶点。