复元醒脑法促进糖尿病脑梗死血运重建中microRNAs的作用

Based on the discovery that microRNAs play a negative regulatory role on gene expression in revascularization for the vascular complications of diabetes and the early clinical and experimental evidences worked by our team,this project intends to reveal the molecular mehanisms and the effective targets of Fuyuan Xingnao Decoction on the revascularization of diabetic cerebral infarction via microRNAs regulating on gene expression in vitro and in vivo.In the experiment, the detection of microvessel density, the ultrastructural changes of cortical microvessels, the cerebral microcirculation, the expression of IGF-1,IGF-1 receptor, CCNE1, CDC25A, the expression of miRNA-503 and miRNA-320 in rats with cerebral ischemia were carried on with erect microcirculation observation system, western blot,real-time fluorescent quantitative polymerasechainreaction (QPCR), RNA interference and other experimental techniques before and after treatment with Fuyuan Xingnao Decoction.The research has an important practical and scientific significance on exploration of the pathogenesis of diabetic vascular complications especially diabetic cerebral infarction as well as the development of the strategies on prevention, treatment and drug discovery.

基于microRNAs在糖尿病血管并发症血运重建中基因表达负性调控的作用，及项目组前期研究证据，本项目拟通过体内、体外实验，采用正立微循环观察系统、Western blot、实时荧光定量PCR、RNA干扰等实验技术和方法，通过对糖尿病脑梗死大鼠新生血管密度检测，脑缺血后皮层微血管超微结构的改变，脑微循环动态变化,IGF-1、IGF-1受体、CCNE1、CDC25A蛋白的表达，miRNA-503和miRNA-320基因表达，以及复元醒脑法对其干预作用的对比研究，并借助RNA干扰技术佐证，以期从microRNAs对基因表达调控作用途径揭示复元醒脑法改善糖尿病脑梗死血管新生的分子机制与有效作用靶点。本研究对深入探讨糖尿病脑梗死等血管并发症的发病机制、制定防治策略、新药研发均具有重要现实与科学意义。

本项目基于microRNA320、 microRNA-503 在糖尿病血管并发症中对血运重建的负性调控作用，揭示糖尿病脑梗死等血管并发症的病理机制，探索复元醒脑汤对糖尿病脑梗死后血管新生的分子机制及具体作用靶点。. 通过体内、体外实验，采用正立微循环观察系统、Western blot、实时荧光定量PCR、RNA干扰等实验技术和方法，观察复原醒脑汤过对糖尿病脑梗死大鼠新生血管密度、脑缺血后皮层微血管超微结构、脑微循环动态变化的影响,对miRNA-503和miRNA-320、IGF-1、IGF-1受体、CCNE1、CDC25A基因及其蛋白的表达的影响，以及复元醒脑法对其干预作用的对比研究，并借助RNA干扰技术佐证，以期从microRNAs对基因表达调控作用途径揭示复元醒脑法改善糖尿病脑梗死血管新生的分子机制与有效作用靶点。. 揭示了复元醒脑汤可以降低糖尿病脑梗死大鼠血糖、血脂水平，改善神经缺损行为体征评分，促进梗死组织区域的血管新生。其可能的分子机制在于复元醒脑汤促进了细胞周期蛋白E1（Cycline1,CCNE1）和CDC25A（cell division cycle 25 A，CDC25A）的表达，抑制了microRNA-503，miRNA-320的表达水平，间接提高了BMECs的成管、迁移和增殖能力，从而使得受损血管内皮修复功能得到改善，进而促进局部血管的再生和侧支循环的建立。本研究在体内、体外分别评价了复元醒脑汤的治疗效果，并对复元醒脑汤的起效机制进行了探究，阐释了复元醒脑汤的作用机理，进一步揭示糖尿病脑梗死的病理本质。为糖尿病脑梗死脑缺血后血管新生的调控途径提供可靠实验依据，具有广阔的运用前景和重要的科学意义。