基于成分群全组合编码—单次测序的消渴安方抗2型糖尿病等效物质群发现研究

Type 2 diabetes mellitus(T2DM) is currently one of the diseases with the highest incidence rate in the world. Traditional Chinese Medicine (TCM), Xiao-Ke-An formula has advantages of “multi-component, multi-target and multi-pathway” to treat T2DM. However, its pharmacodynamic substances are unknown. Recently，multiple research teams have proposed a series of methods on the study of the discovery of pharmacodynamic substances group of TCM, such as “bioactive equivalent combinatorial components”, “target component knockout and knock-in”, “network overall regulation”. These work have made good progress, but they all have a problem with insufficient throughput. In this study, we will focus on the scientific issue of identifying the equivalent substance group of Xiao-Ke-An formula, introducing the idea of barcoding, combining component knockout strategy and RNA-seq, establishing a new model for the discovery of pharmacodynamic substances group based on complete combination of components barcoding RNA-seq (tcbRNA-seq). it is expected to break through the technical bottlenecks with limited throughput, and significantly improve the throughput and efficiency on discovery of pharmacodynamic substances of TCM. This study can clarify the equivalent substance group of Xiao-Ke-An formula treatment of T2DM, and it is also expected to establish a universal new model for discovery of the pharmacodynamic substances with low-cost and high-throughput, which will help to promote the modernization of traditional Chinese medicine.

2型糖尿病目前是全世界发病率最高的疾病之一。中药复方消渴安方治疗2型糖尿病具有“多成分、多靶点和多途径”整体调节的优势，但其药效物质尚未阐明。近年来，多个研究团队提出“等效成分群”、“目标成分敲除/敲入”和“网络整体调控”等中药药效物质群发现方法，取得了良好进展，但这些方法均存在通量不足的瓶颈问题。本项目拟围绕消渴安方抗2型糖尿病等效物质群这一科学问题，引入编码的思路，结合成分敲出和转录组测序等技术，建立基于成分群全组合编码—单次测序（complete combination of components barcoding RNA-seq，tcbRNA-seq）的药效物质群发现新模式，有望突破通量受限的技术瓶颈，大幅提升当前药效物质群发现策略的通量和效率。本研究可阐明消渴安方抗2型糖尿病的等效物质群，也有望建立一种具有普适性的低成本、高通量的中药药效物质发现新模式，有助于推动中药现代化。

2型糖尿病目前是全世界发病率最高的疾病之一。中药复方消渴安方治疗2型糖尿病具有"多成分、多靶点和多途径"整体调节的优势，但其药效物质尚未阐明。近年来中药药效物质研究方法存在通量不足的瓶颈问题。本项目开发了一个基于特异性cDNA标签编码的多重样本平行转录组测序技术（Transcriptome Changes for Multiplexed samples with RNA-seq，TCM-seq），具有高通量、低成本等优势，可用于评估中药等复杂体系的药效作用与分子机制研究。并通过构建多种糖尿病相关体外细胞模型和转录组测序，明确消渴安方中“君药”黄连抑制糖异生和脂肪生成的药效物质与作用机制，并在胰岛素抵抗的肥胖小鼠中进行体内验证。我们的研究结果为中药药效物质和作用机制研究提供了新的方法，有助于推动中药现代化。