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改善血流、促进血管新生是缺血性外周血管疾病的重要治疗措施.由于载脂蛋白A(ApoA)与纤溶酶原(plasminogen,Plg)具有75%～98%的结构同源性,因此,ApoA也可能通过类似Plg的方式抑制内皮祖细胞(endothelial progenitor cells,EPCs)增殖、黏附及迁移而影响血管发生的能力.本文研究ApoA对EPCs血管发生的影响及机制.为了编码人ApoA全长cDNA序列的pSG-5表达载体,转染COS-7细胞株后进行培养,收集培养液,免疫亲和层析法分离纯化ApoA蛋白;从转ApoA基因小鼠、野生型对照鼠及正常对照鼠骨髓分离培养EPCs,经ApoA处理后移植下肢缺血实验小鼠,于移植后第3、7、14天后观察ApoA对EPCs黏附、迁移及血管发生能力的影响.研究发现,ApoA能显著降低EPCs的黏附、迁移能力,Matrigel胶上,EPCs血管腔样结构严重破坏,体内实验揭示,EPCs归巢至ApoA转基因小鼠缺血组织血管周围的数量及毛细血管数量显著减少.结果表明,ApoA能损伤EPCs的黏附、迁移及归巢,最终损伤EPCs的血管发生能力.
Improve blood flow and promote angiogenesis is an important treatment of ischemic peripheral vascular disease.As apoA (ApoA) and plasminogen (plasminogen, Plg) has 75% to 98% of the structural homology, so , ApoA may also inhibit the proliferation, adhesion and migration of endothelial progenitor cells (EPCs) to affect the angiogenesis through a similar Plg way.In this paper, the effect of ApoA on EPCs angiogenesis and its mechanism were studied.In order to encode human ApoA full-length cDNA sequence of pSG-5 expression vector, transfected COS-7 cell line after culture, collection of culture medium, immunoaffinity chromatography ApoA protein isolated and purified; from transgenic ApoA mice, wild-type control and normal control mice EPCs were isolated from bone marrow and treated with ApoA, then the mice were sacrificed on the 3rd, 7th and 14th day after transplantation to observe the effect of ApoA on the adhesion, migration and angiogenesis of EPCs.The study found that ApoA can significantly reduce the EPCs Adhesion, migration ability, Matrigel gel, EPCs vascular cavity-like structure of the serious damage, in vivo experiments revealed that EPCs homing to ApoA transgenic mice peri-ischemic vascular volume and the number of capillaries significantly reduced. Ming, ApoA can damage EPCs adhesion, migration and homing, and ultimately damage EPCs angiogenesis.