基于肠道微生态研究黄芩-黄连药对干预2型糖尿病的“相须”配伍机制

Based on our previous studies and the close relations between intestinal microecology and type 2 diabetes, the research idea is proposed that the treatment of diabetes by the scutellaria-coptis herb couple is obtained through the regulation of intestinal microecology, intervention of inflammation and insulin signaling pathways and regulation of glucose and lipid metabolism. And the following studies will be carried out: (1) The uniform design method and biological effect evaluation method will be applied to investigate the dose-effect relationship of the herb couple compatibility; (2) Based on the metagenomics methods, the regulation of the herb couple compatibility on the biodiversity and function of intestinal flora in the type 2 diabetes animals will be analyzed; (3) Based on the metabolomics and molecular biology methods, the effects of the herb couple compatibility on biomarkers, the key target proteins of inflammation and insulin signaling pathways, key enzymes of glucose and lipid metabolism pathways and islet β cell function will be investigated. The final objects of these studies are to systematically clarify the synergistic compatibility mechanism of the herb couple for the treatment of type 2 diabetes through regulating intestinal microflora, which will contribute to its effectively clinical application.

基于肠道微生态与2型糖尿病密切相关的现代认识以及前期研究积累，提出：黄芩-黄连药对通过调控肠道微生态，影响炎症及胰岛素信号通路，调节糖脂代谢，实现有效干预2型糖尿病的工作思路。拟开展以下研究：（1）应用均匀设计法及生物效应评价方法研究药对配伍的量-效关系；（2）应用宏基因组学方法分析药对配伍对模型动物肠道菌群多样性及功能的调控作用；（3）应用代谢组学和分子生物学方法分析药对配伍对模型动物生物标志物、炎症及胰岛素信号通路关键靶标蛋白、糖脂代谢途径关键酶及胰岛β细胞功能的影响。以期阐明黄芩-黄连药对通过调控肠道微生态干预2型糖尿病的“相须”配伍机制，揭示黄芩-黄连药对传统中医“止消渴”的科学内涵，为提高临床用药有效性提供参考。

黄芩-黄连药对干预2型糖尿病的协同增效作用已被临床实践所证实，但配伍机制目前仍缺乏合理阐释。本项目基于肠道微生态紊乱与糖尿病密切相关的现代认识，开展黄芩-黄连配伍调控肠道微生态，影响炎症及胰岛素信号通路，调节糖脂代谢的研究。采用高糖高脂饲料喂养，联合注射低剂量STZ，建立T2DM大鼠模型，并给予不同配比、不同剂量黄芩黄连药对提取液，结果表明，黄芩-黄连比例为3:2~1:3时，具有显著的协同作用，且1:1高剂量组协同效应较强。黄芩-黄连配伍显著提高短链脂肪酸产生菌的相对丰度，显著改善T2DM大鼠肠道菌群代谢紊乱，提高乙酸、丙酸、丁酸等短链脂肪酸、结合型胆汁酸如甘氨胆酸、甘氨脱氧胆酸的含量，降低游离型胆汁酸如胆酸、脱氧胆酸的含量。此外，黄芩-黄连配伍显著抑制MAPK信号通路关键靶点NF-ƙB/P65磷酸化水平及血浆中CRP、IFN-γ、TNF-α、IL-6、resistin、SOCS3、IL-1β、NO的表达，显著提高胰岛素信号通路关键靶点蛋白PI3K、Akt磷酸化水平以及GK、PFK、PK、GS的酶活性，显著抑制G6Pase、PEPCK、FBPase、GP、HMG-CoA、ACC、HSL、FAS的酶活性。黄芩-黄连配伍显著改善内源性生物标志物的代谢紊乱，降低T2DM大鼠血浆中胆酸、脱氧胆酸等以及尿液中黄嘌呤核苷、脱氧尿苷、D-葡糖醛酸-6,3-内酯等含量；提高T2DM大鼠血浆中溶血磷脂酰胆碱、溶血磷脂酰乙醇胺、磷脂酰胆碱、甘氨胆酸等以及尿液中马尿酸、N-乙酰-L-酪氨酸等含量。综上所述，本项目从量效关系、肠道微生态及代谢组学等方面揭示了黄芩-黄连干预T2DM的配伍机制。在项目执行过程中，发表学术论文11篇（标注基金号），其中SCI论文6篇，获得授权专利1项。本项目的研究成果为黄芩-黄连配伍干预T2DM的临床应用提供了科学依据。