养阴益气活血法对糖尿病大血管病变内质网应激维稳机制研究

Promoting blood circlulation in preventing and controling diabetic macroangiopathy has been paid much attention in recent years. However, the benefits of using promoting blood circlulation with no obvious signs of blood stasis in the early treatment of diabetic macroangiopathy is uncertainty. In our last natural fund research, we used promoting blood circlulation with no obvious signs of blood stasis in the early treatment of diabetic macroangiopathy. And we found that Shenqi compound recipe with efficacy of tonifying Qi and Yin and promoting blood circlulation can delay atherosclerosis occurrence and development. In addition, the study also found that the effect of simple tonifying Qi and Yin is no perfect. The composition of tonifying Qi and Yin is more relative with hypoglycemia,while the composition of promoting blood circlulation is more relative with anti-atherosclerotic effects. We assume that tonifying Qi and Yin and promoting blood circlulation is an important and effective therapeutic method of Chinese medicine. Promoting blood circlulation plays a key role in keeping homeostasis of diabetic macroangiopathy through interventing endoplasmic reticulum stress signal pathway.In this study, we decompose Shenqi compound recipe into two recipes: tonifying Qi and Yin recipe and promoting blood circlulation recipe .We will compare the regulation of Shenqi compound recipe and its decomposed recipes on reticulum stress signal pathway . We try to compose Shenqi compound recipe and its decomposed recipes in regulating endoplasmic reticulum stress signal network, and analyzes the connection of molecular network.We try to explore the importance and scientific of the early intervention of promoting blood circlulation in preventng and control of diabetic macroangiopathy, and we try to figure out the preliminary mechanismof promoting blood circlulation in the prevention and treatment of diabetic macroangiopathy.

活血法在防治糖尿病大血管病变中的作用已受到广泛重视，然而没有明显血瘀征象早期活血治疗是否有益尚无定论。我们之前的国家自然基金研究，在未出现瘀血症状时即采用活血法，发现具有养阴益气活血功效的参芪复方可防治糖尿病大血管病变的发生发展；此外研究还发现，单纯养阴益气效不佳，养阴益气组分主要与降糖效应相关，而抗动脉粥样硬化则主要与活血组分相关。我们推测"养阴益气活血法是防治糖尿病大血管病变重要及有效治法，活血法的早期应用可通过调控内质网应激以维持糖尿病大血管病变机体内环境稳定而起到关键作用"。本次研究将参芪复方拆为养阴益气组分及活血组分，对比参芪复方及其拆方组分对主动脉内质网应激的调节作用，并尝试构全方及各组分调控内质网应激的信号网络，分析分子网络联系，探索活血法在早期介入防治糖尿病大血管病变中的重要性及应用的科学性，探索活血法早期应用于防治糖尿病大血管病变的初步机制。

本项目围绕工作假说“养阴益气活血法是防治糖尿病大血管病变重要及有效治法，活血法的早期应用可通过调控内质网应激以维持糖尿病大血管病变机体内环境稳定而起到关键作用”，以具有养阴益气活血功效的参芪复方为载体，通过对参芪复方及其活血组分早期运用的研究，旨在从整体水平、细胞水平、分子水平初步探讨参芪复方中活血组分早期运用的合理性与科学性，为在糖尿病大血管病变的防治中早期使用活血法提供理论依据。将参芪复方拆分为养阴益气组分和活血组分，对比参芪复方及其拆方组分对KKAy小鼠一般情况、空腹血糖、血脂水平、超敏 C 反应蛋白，HE 染色对腹主动脉进行形态学观察，应用 Agilent miRNA 芯片技术平台对腹主动脉组织的 miRNA 表达情况进行检测，筛选出差异表达 miRNAs；并利用 TargetScan 数据库预测其靶基因，进一步对靶基因进行 GO 和 Pathway 等生物信息学分析，以深入了解 miRNAs 的调控机制和生物学功能，逐步明确了全方及各组分对内质网应激信号网络的调控作用，结果表明参芪复方中活血组分的早期运用可以改善糖尿病大血管病变 KKAy 小鼠的一般状态，改善脂代谢紊乱，降低 hs-CRP，减轻模型小鼠血管内皮损伤，延缓糖尿病动脉硬化的进展，其作用机理可能是通过调控生物学过程中细胞命运的确定、改善应激反应等，从而防治糖尿病大血管病变。因此，本研究结果证明了活血法在早期介入防治糖尿病大血管病变中的重要性及应用的科学性，为活血法早期应用于防治糖尿病大血管病变提供一定的理论基础。本研究目前发表了1篇中文核心期刊论文及1篇SCI 论文（录用待刊出），由于实验数据庞大，分析复杂，正式实验结果虽已整理，暂时只有1篇论文在审稿当中，尚有论文正在分析成文，准备投稿中，培养了3名硕士研究生。