基于肠道微生态和代谢组学的益智治疗慢性肾损伤的物质基础与分子机制研究

Alpinia oxyphylla is mainly distributed in Hainan province and has the effect of treatment of renal disease and reduce the urinary production activity. It commonly used in the therapeutic of chronic kidney disease (CKD), which have been confirmed by the reports and our previous studies. Recently research shows that the CKD is closely related to alterations the intestinal microflora. The toxicants absorbed from the gut induced the kidney inflammation and oxidation which lead to the fibrosis of the renal tubule. Accordingly, based on the Chinese medicine theory, the project proposed to study the bioactive components and molecular mechanism of Alpinia Fructus. The following works will be carried out: (1) The metabolite profiles of CKD animal models will be established using the metabolomic methods and techniques, and the biomarkers and metabolic pathway by the chemical composition in Alpinia Fructus will be studied. The presumable action mechanics will also be speculated; (2) Based on the molecular biology techniques, an intestinal microflora fingerprint of CKD animal models will be established, and the action of the chemical composition of Alpinia Fructus on animal model’s intestinal flora will also be studied; (3) Based on the results of metabonomics and intestinal microflora, the action mechanism of bioactive components of Alpinia Fructus on CKD were verified using modern pharmacologic models. The final objects of these studies are to systematically clarify the bioactive components and the biological mechanism of Alpinia Fructus for the remedy of CKD and to reveal the scientific connotation of its anti-diuretic effects.

益智为海南省地产道地药材，具有暖肾缩尿的功效，广泛应用于慢性肾病（CKD）的治疗和预防。研究发现肠道微生态异常，导致毒性物质进入血液循环，诱导肾脏氧化应激和肾小管纤维化，从而形成CKD。基于 “多尿是肾脏损伤早期的一种表现，益智通过调节肠道微生态，缓解肾脏氧化应激和纤维化其中一个或多个环节，减轻肾脏损伤而实现暖肾缩尿的功效”的思路。拟开展研究工作：（1）运用代谢组学方法和技术研究益智不同化学部位干预腺嘌呤肾损伤和糖尿病肾损伤模型动物中肠肾相关代谢物谱的效应、调控靶标、影响代谢通路等，探讨其作用途径；（2）基于肠道微生态研究方法，构建肾损伤模型动物肠道菌群结构和指纹图谱，研究益智不同化学部位对肠道微生态的调节作用；（3）通过相关靶细胞模型，对益智活性部位及其主要效应成分进行效应评价及分子机制研究。以期揭示益智对CKD的整体调节作用与分子机制，阐明益智传统中医“暖肾缩尿”的功效特点和科学内涵。

中药益智为海南省道地药材，具有补肾缩尿的功效，在临床上对慢性肾损伤导致的多尿症有较好的疗效，但是由于其物质基础不明确，制约了益智健康产品的开发。本课题组前期实验证实益智主要含有倍半萜、二苯基庚烷和黄酮类物质，并从中鉴定了11种单体化合物。 本项目拟采用代谢组学和肠道菌群调节等新技术新方法对益智治疗肾损伤的物质基础进行研究。采用现代分离方法获得益智倍半萜部位和二苯基庚烷、黄酮部位，以肾损伤动物模型对不同部位的药理活性进行评价，发现倍半萜对肾损伤具有较好的保护作用。代谢组学研究发现，益智倍半萜具有较好的降低肾毒性物质和减轻肾脏纤维化的作用，其代谢途径与调节肠道平衡和减轻肾脏炎症有关。肠道微生态研究发现，益智倍半萜可以增加肠道菌群数量，减轻肠道炎症。HK-2细胞实验证明益智倍半萜和诺卡酮可以调节细胞活性氧的产生，抑制细胞纤维化，进而减轻肾组织损伤。本项目为益智的开发利用提供科学依据，为海南省中药现代化发展和益智临床应用提供了技术支持。