基于肠道菌群及其代谢的交泰丸治疗糖尿病配伍机制研究

The occurrence of diabetes is closely related to the gut microbiota. Jiaotai Pill（JTP） is an effective formula for the treatment of diabetes. However, it has not been elucidated how JTP can produce therapeutic effects and its compatibility mechanism by regulating gut microbiota. Based on the previous studies, this study used gut microbiota of type 2 diabetes (T2DM) model rats as the starting point, based on the evaluation of the therapeutic effect of different ratios of JTP on T2DM model rats through pharmacological indexes such as glucose metabolism, diversity of gut microbiota were analyze using 16S rRNA method, and the improvement of gut microbiota structure with different ratios of JTP was determined. Metabolomics methods were used to identify and confirm metabolic biomarkers of gut microbiota in T2DM rats, to reveal the effects of JTP on metabolism of gut microbiota. Then, co-variation analysis was conducted on intestinal flora diversity and metabolic biomarkers, and associated functional bacteria and metabolic biomarkers were found. On this basis, correlation analysis was performed with the indicators of glucose metabolism and so on. From the perspective of gut microbiota and its metabolic dynamics, clarifying the compatibility mechanism of JTP in the treatment of T2DM with multiple targets, promoted the application of JTP in diabetes, and explored a new method for the study of the compatibility mechanism of formula, which provide new ideas for enriching and developing theoretical research on compatibility of traditional Chinese medicine prescriptions.

糖尿病的发生与肠道菌群关系密切，交泰丸治疗糖尿病具有坚实的理论和应用基础，临床疗效确切，但其如何通过调节肠道菌群变化产生治疗作用及其配伍机制尚未阐明。在前期研究基础上，本研究以2型糖尿病（T2DM）模型大鼠肠道菌群失调为切入点，在通过血糖、血脂等药理指标评价不同配伍比例交泰丸对T2DM治疗作用基础上，应用16S rRNA方法分析肠道菌群多样性，明确交泰丸对T2DM大鼠肠道菌群结构的干预作用及不同配伍变化规律；采用代谢组学方法，发现并鉴定T2DM模型大鼠肠道菌群代谢生物标记物，分析相关代谢通路，揭示交泰丸对肠道菌群代谢的影响及配伍变化；最后，将肠道菌群多样性和代谢生物标记物进行共变化分析，发现关联功能菌与代谢生物标记物，并与血糖、血脂等指标进行关联，从肠道菌群及其代谢动态关联角度，阐明交泰丸治疗T2DM的多靶点整体效应配伍作用机制，为丰富和发展方剂配伍理论提供新的思路，促进中医药继承和发展。

交泰丸为经典方剂，目前临床上用于治疗糖尿病，疗效确切，但其是否通过调节与糖尿病密切相关的肠道菌群变化而产生治疗作用以及其配伍机制为何，尚未阐明。本项目以优化后的高脂饮食+小剂量多次注射链脲佐菌素大鼠为模型，首先对交泰丸及黄连-肉桂不同比例配伍治疗T2DM的治疗作用进行了评价，交泰丸对T2DM模型大鼠的血糖值、HbA1c、TC、TG、LDL、IL-6、IL-10等药理指标的具有较好的调节作用，随黄连-肉桂中肉桂比例的升高，作用强度逐渐降低，10:5比例仅对个别指标具有调节作用，但缺肉桂配伍组的作用不及交泰丸组；应用16S rRNA方法分析肠道菌群多样性，T2DM模型大鼠的4个门类、6个科、16个属的菌群发生变化，验证了糖尿病和肠道菌群的密切关系，交泰丸给药组可对4个门、4个科、14个属的菌群起到调节作用，黄连-肉桂不同比例给药组也不同程度的对紊乱的肠道菌群有的回调作用；采用代谢组学方法，发现并鉴定T2DM模型大鼠肠道菌群代谢生物标记物21个，交泰丸及黄连-肉桂不同配伍比例组均可对标记物进行回调，其中交泰丸可回调20个，黄连-肉桂10:3组回调了18个，10:5组17个，10:7组16个，黄连组17个，肉桂组10个。综合分析，黄连-肉桂配伍中，黄连比例越高对糖尿病的治疗、肠道菌群多样性调节和代谢标记物回调作用越显著，比例为10:1即交泰丸古方比例时为最佳。研究从药理作用、肠道菌群多样性以及肠道菌群代谢多个角度阐释了交泰丸配伍的科学性和合理性，为丰富和发展方剂配伍研究提供新的思路和依据。