芪丹地黄颗粒治疗糖尿病肾病的基因表达调控网络机制研究

Diabetic nephropathy is one of the most common complications of diabetes, the mechanism of which covers a wide range and cannot be clearly defined yet. The applicant mainly focuses on the mechanism functioning on the disease, and find that the onset of diabetic nephropathy is contributed by the changes of multi-genes and multi-targets instead of the sole target. While the fact has been universally acknowledged that traditional Chinese medicine works on more than one target in many ways, it remains a difficulty to do further research on its complex mechanism. This program make a tentative attempt to intervene diabetic nephropathy by using Qi dan Di huang Granule, during which gene regulation network method helps a lot in selecting the co-gene between diabetic nephropathy and Qi dan Di huang Granule, detecting the changes through gene expression and spotting the target gene of Qi dan Di huang Granule affecting the disease. In correspondence to its effective constituents, tests ,In vivo and in vitro , have been made to confirm the effect on diabetics resulting from Qi dan Di huang Granule and its effective constituents, thus analyzing its functioning mechanism, identifying its effective constituents, and further optimizing the structure of Qi dan Di huang Granule. This has provided a new method for a better understanding of diabetic nephropathy and researches concerning Chinese Traditional Medicine Compound.

糖尿病肾病是糖尿病最常见的并发症之一，其内在机制涉及诸多方面，确切机制尚不清楚。申请人专注于糖尿病肾病基因表达调控网络变化对糖尿病肾病的影响，在研究中发现糖尿病肾病发病涉及多基因参与，单一靶点干预难以取得理想效果。中医药复方制剂能够作用于多基因、多靶点已得到广泛认同，但对其复杂作用机制的研究尚是难点。本项目拟运用治疗糖尿病肾病的循证处方——芪丹地黄颗粒，通过对基因调控网络的分析，筛选出芪丹地黄颗粒与糖尿病肾病之间共同基因，检测共同基因表达谱变化，找出糖尿病肾病中受芪丹地黄颗粒影响变化的基因，对应其可能有效作用成份，通过体内、体外模型，多角度验证芪丹地黄颗粒及其有效成份对糖尿病肾病基因表达谱的影响，解析其发挥效应的机制，鉴定其作用的有效成份，进一步优化组方结构，这为糖尿病肾病机制的认识以及中药复方机制的研究提供新的思路与方法。

糖尿病肾病是糖尿病最常见的并发症之一，发病机制涉及遗传、环境等诸多因素，西医治疗未能完全阻止其进展。我们的前期研究发现芪丹地黄汤在缓解糖尿病糖尿病肾病的尿蛋白、肾功以及临床症状方面有确切疗效，但中药复方成分及其靶点的多样性、糖尿病肾病作用机制的复杂性使得中药复方治疗疾病作用机制的研究成为难点。本项目首先通过网络药理学方法筛选芪丹地黄汤中的活性成分和治疗糖尿病肾病的靶点，并预测芪丹地黄汤在系统、器官和分子水平上的药理学机制，其次进一步通过动物及细胞实验多角度验证芪丹地黄汤及其有效成梓醇对糖尿病肾病基因表达的影响。结果表明芪丹地黄汤可能通过复杂的网络机制调节来发挥延缓糖尿病肾病的进展，包括葡萄糖和脂质代谢、炎症反应、对氧化应激和缺氧的应答、纤维化等过程，以及RAS，TGF-β， NF-κB，PI3K-Akt-mTOR和HIF-1等信号传导途径。动物及细胞研究发现芪丹地黄汤改善糖尿病肾病的作用与调节脂质代谢、肾纤维化、炎症反应以及肾小管上皮细胞EMT相关。本项目进一步确定了芪丹地黄汤对糖尿病肾病的疗效，并且初步明确其作用机制，为其临床应用提供依据。