痛风脾虚湿阻证肠道菌群结构与功能特征及祛浊通痹方干预机制研究

Gout has become a common and frequently occurring disease which has major impact on patients' living quality and health. The latest scientific research suggests that decreased extra-renal urate excretion is a common cause of hyperuricemia and gout, which has been neglected for a long time. Intestinal flora is the main environment of uric acid decomposition in the gut because of its urate oxidase. Our previous research showed that spleen deficiency and dampness syndrome is the main one for the gout in our country, and this kind of patient has unique characteristics of intestinal flora, along with obvious disorder of flora structure and function. Therefore, we speculate that the gut dysbiosis played an important role in the pathogenesis of gout-Spleen deficiency and dampness syndrome, while "Qu- Zhuo- Tong- Bi" Recipe may play its role by adjusting the structure and function of gut microflora. We intend to study the structural and functional characteristics of gut microflora in the means of 454 high-throughput sequencing and real-time fluorescence quantitative PCR technology, combining with bioinformatics analysis to screening the syndromes microecological molecular and detecting differences of uricase gene abundance so as to discuss the impact of "Qu-Zhuo-Tong-Bi" Recipe on the structure and function of the intestinal flora. This research will contribute to the etiology and pathogenesis of gout-Spleen deficiency and dampness syndrome from a new perspective, as well as provide new ideas and targets for the prevention and cure with TCM.

痛风已成为一种常见病和多发病，严重影响患者的生活质量及生命健康。最新科学研究表明，一直以来被忽视的肠道排出尿酸功能的降低是导致高尿酸血症及痛风的一个重要原因，而肠道菌群因具有尿酸酶而成为尿酸在肠道分解的主要环境。课题组前期跟踪研究发现脾虚湿阻证为我国主要的痛风证候类型，且此证型患者具有独特的肠道菌群特征，存在明显的菌群结构及功能紊乱。因此，我们推测肠道微生态失衡在痛风脾虚湿阻证的发病中发挥了重要的作用，而祛浊通痹方可能通过调整肠道微生态结构及功能以发挥作用。我们拟采用454高通量测序及实时荧光定量PCR技术，结合生物信息学分析，研究痛风脾虚湿阻证的肠道菌群结构与功能特征，筛选病证相关微生态分子，检测尿酸酶基因丰度差异，探讨祛浊通痹方对肠道菌群结构与功能的影响。本课题将从新的视角阐明原发性痛风脾虚湿阻证的病因病机，并为痛风脾虚湿阻证的中医药预防和诊疗提供新思路和新靶点。

痛风是嘌呤代谢紊乱、尿酸排泄障碍所致血尿酸增加的一种常见病和多发病，严重影响患者的生活质量及生命健康。最新科学研究表明一直以来被忽视的肠道排出尿酸功能的降低是导致高尿酸血症及痛风的一个重要原因。本研究通过能无创获得、更容易为受试者所接受的粪便样本，结合Illumina Miseq高通量测序技术及高灵敏度核磁共振谱技术，系统研究了能深入反映肠道菌群与宿主相互作用的粪便代谢指纹图谱和肠道菌群结构特征，筛选获得了痛风患者的特征性微生物标志分子及特异性代谢标志物。研究结果表明，痛风人群中，Bacteroidetes 中的多个微生物物种明显富集而Chloroflexi中的多个微生物种类则显著下调。通过粪代谢组学研究发现丙氨酸、甘氨酸、琥珀酸、乙酸、牛磺酸、肌苷、-木糖、-葡萄糖和-葡萄糖等9种代谢物在痛风患者中上调，而缬氨酸、天冬酰胺、天冬氨酸、瓜氨酸、苯丙氨酸和-酮异戊酸等6种代谢产物则表现为下调。关联分析表明肠道菌群结构变化可影响宿主代谢过程，而宿主代谢产物水平的变化在一定程度上可调控肠道菌群结构的组成。此外，实验结果表明祛浊通痹方可调控肠道菌群结构，可能通过这一机制来发挥相应的作用。本研究在一定程度上揭示了痛风患者体内尿酸排泄、嘌呤代谢紊乱和炎症因子等过程的变化，探讨了代谢产物变化与肠道菌群之间的相关性及两者在痛风发生发病中的可能机制，建立了符合原发性痛风的宏基因组学及代谢组学诊断、判别模型，并从代谢组学水平上探讨了痛风中医“证”的物质基础，对中医证型客观化、标准化研究进行了有意义的探索，丰富了中医证候的科学内涵。脾虚湿阻证