藏医名方中有效单体挖掘及其诱导EPCs调控脑缺血后血管再生修复的研究

Endothelial progenitor cells (EPCs) are of critical important in local angionenesis, blood supply increase, ischemic injury decrease and corresponding prognosis. EPCs exists of a very limited amount in normal body. While in vitro proliferation of EPCs is commonly detected to bear the risk of phenotypic variation, over differentiation and fading. Detection of an inducing chemical to increase the amount of EPCs is a prospect in treatment of cerebral ischemia.. In the present study, promoter probe eukaryotic expression plasmid pSVrVEGFpro-Luc transfected Hek293 cell is established and screened to obtain a cell strain with stable expression of pSVrVEGFpro-Luc promoter plasmid. The cell stain is then used to screen the target active ingredient from the selected 17 chemicals obtained from the three classic Tibetan medicines. Based on the similarity of the therapeutic principle between ‘Stem Cell Self-Renewing’ and ‘Activating Blood Circulation to Dissipate Blood Stasis’, it is assumed that some target active ingredient may activate endothelial progenitor cells and promote angiogenesis. The mechanism of repair and regeneration of artery will be investigated in vitro and in vivo to elucidate the therapeutic efficacy of classic Tibetan medicines on cerebral ischemia.

内源性EPCs对缺血组织局部血管新生、血供增加、减轻缺血损伤及其预后至关重要，但正常机体内EPCs数量较少，而体外扩增弊端重重，故寻求高效能的化学诱导物增加体内EPCs数目成为脑缺血治疗的一大前景措施。 在本项目中，我们首先利用前期成功构建的启动子探针型真核表达质粒pSVrVEGFpro-Luc转染Hek293细胞，筛选得到稳定表达pSVrVEGFpro-Luc启动子质粒的稳定细胞株；用该模型对从三种藏医名方入血成分中追踪到的17种单体进行筛选，快速、高通量地挖掘出其中促进血管新生的有效单体。结合“干细胞循环”与“活血化瘀理论”存在的诸多相似性，大胆推测：其中某些单体成分极有可能通过激活骨髓EPCs，参与缺血局部组织的血管再生修复过程。为此，我们将从细胞生物学和分子水平揭示该单体修复或重建血管网络的机制，并在疾病动物模型体内进行药效验证，为深入挖掘藏成药抗脑缺血药效物质基础提供新思路。