基于“肠道菌群-炎症-胰岛”轴研究藏药小檗皮治疗“京尼萨库”病的药效物质及作用机制

Berberidis cortex is a commonly used Tibetan medicine for the treatment of “Jingnisaku” disease (i.e., diabetes). In our previous studies, Berberidis cortex has been found to contain several alkaloids (NSFC 81303310), and have obvious anti-diabetic effects (NSFC 81173360). However, its effective substances in vivo and the mechanism of action are unknown. According to pathological features during the pathogenesis of diabetes, such as gut microbiota imbalance, inflammatory reaction, and insulin resistance, this project proposes a hypothesis: “After the oral administration of Berberidis cortex, the metabolites of the gut microbiota and the ingredients that are absorbed into the blood are the effective substances for the treatment of diabetes, and the regulation of "Gut microbiota-Inflammation-Pancreatic islet" (GIP) axis is its mechanism of action.” This study is planned to integrate the use of multidisciplinary techniques and methods, such as intestinal microbial metabolism, metagenomics, and metabolomics, to study the metabolites of gut microbiota in vivo and in vitro and the ingredients absorbed by the intestines into the blood. Combined with pharmacodynamic validation, it can reveal the effective substances of Berberidis cortex. On the other hand, it is planned to study the effects of Berberidis cortex on intestinal microflora, inflammatory factors, short-chain fatty acids, insulin, insulin resistance index, GLP-1, and endogenous metabolites in diabetic rats' GIP axis. By exploring the relationship between its anti-diabetic effects and changes in intestinal microflora and biomarkers, it is possible to reveal the mechanism of action of Berberidis cortex in the treatment of “Jingnisaku” disease.

小檗皮为治疗“京尼萨库”病（糖尿病）的常用藏药材。课题组前期发现小檗皮主要含有生物碱类成分（NSFC 81303310），并具有明显的抗糖尿病作用（NSFC 81173360），但体内药效物质及作用机制不明。本项目针对糖尿病发病过程中常出现肠道微生物失衡、炎症反应、胰岛素抵抗等病理特征，提出“小檗皮口服后其肠道菌群代谢与吸收入血产物是其治疗糖尿病的药效物质基础，而调节肠道菌群-炎症-胰岛轴（GIP轴）是其作用机制”的假说。拟整合运用肠道微生物代谢、宏基因组学、代谢组学等技术及方法，研究小檗皮生物碱成分体内外肠道菌群代谢、肠吸收入血产物，结合药效学验证，揭示其体内药效物质基础；研究小檗皮对糖尿病大鼠GIP轴中肠道微生物、炎症因子、短链脂肪酸、胰岛素、胰岛素抵抗指数、GLP-1、内源性代谢物的影响，探讨其抗糖尿病效应与肠道菌群变化及生物标志物间的联系，揭示小檗皮治疗“京尼萨库”病的作用机制。

藏药小檗皮为治疗“京尼萨库”病（2型糖尿病）的常用药材，疗效确切，但其药效物质基础和作用机制不明，影响其开发利用。本项目整合应用现代多学科技术和方法，从体内化学成分与肠道微生物相互作用的角度入手，揭示了小檗皮治疗2型糖尿病的体内药效物质和作用机制。（1）对小檗皮药材进行了“化学成分分离-定性定量分析-质量标准建立-质量多维评价”系统研究，首次分离得到蟾毒色胺内盐和阿魏酸4-O-β-D-吡喃葡萄糖苷，建立了规范化的质量标准、一测多评和指纹图谱方法，提高了药材的质量控制水平。（2）采用高脂饲料联合低剂量链脲佐菌素建立2型糖尿病大鼠模型，基于“肠道微生物代谢-体内入血成分-计算机筛选”研究策略，利用UPLC-Q-Exactive Orbitrap MS定性和HPLC-QqQ-MS定量分析技术，发现小檗皮治疗2型糖尿病的体内药效物质主要为蟾毒色胺、蟾毒色胺内盐、去亚甲基小檗碱、芬氏唐松草定碱、小檗碱和小檗红碱。（3）采用16S rRNA高通量测序、代谢组学、分子生物学等研究技术和方法，结合伪无菌大鼠模型，发现小檗皮可通过调控“肠道菌群-炎症-胰岛”轴而改善2型糖尿病，即明显改善肠道菌群紊乱、减轻炎症和胰岛素抵抗、提高胰岛素敏感性、改善糖代谢和氨基酸代谢。Akkermansia、肠球菌、梭杆菌等肠道微生物可能为小檗皮治疗2型糖尿病的关键细菌。（4）首次发现含小檗皮复方制剂“八味小檗皮胶囊”具有改善2型糖尿病的作用，能显著减轻胰岛素抵抗、减少炎症及降低血脂水平，扩大了临床适应症。本项目取得了较好的研究成果，共发表学术论文14篇，其中SCI论文7篇，CSCD论文6篇；申请国家发明专利1项；培养硕士研究生5人；制定四川省地方标准1项；相关研究成果获得中国民族医药学会科技进步奖一等奖1项、四川省科技进步奖二等奖1项、中华中医药学会科学技术奖二等奖1项。本项目研究结果可为小檗皮药材的临床合理应用、质量控制与现代化开发利用提供科学依据，也可为口服藏药体内药效物质基础及起效机制等关键问题研究提供新的思路和方法学参考。