The basic research and clinical trial of therapeutic angiogenesis using bone marrow implantation with EPO

A.骨髓移植（BMI）联合促红细胞生成素（EPO）注射液的临床试验（1）BMI单药治疗及BMI与EPO 6,000 IU联合治疗的结果：EPO注射液未引起重大副作用，联合治疗与单药治疗相比未观察到额外作用。我们报道了植入的CD 34阳性细胞的数量是影响BMI临床疗效的主要因素之一。(Circ J 2004;68：1189-93）（2）BMI与大剂量EPO 24，000 IU联合治疗的结果：4例患者参与。1例1年前接受过单药治疗的患者再次接受高剂量联合治疗。踝臂压力指数的变化显示了高剂量组合的优势：单药治疗，0→0，高剂量组合，0→0.67。然而，其他3名患者显示出高剂量EPO的任何额外作用。然后我们尝试了一种组合， 关于我们 降低BMI和序贯施用EPO以获得EPO在缺血器官中的延长作用。(3)BMI和EPO 6，000 IU x 5天联合治疗的结果：4名患者参与。其中一人显示出休息疼痛的改善，但其他人没有，并遭受截肢，后来。对于这些严重的病人，需要进一步的联合治疗。红细胞和EPO促血管生成机制的基础研究（1）EPO对BMI的补充作用（体内）：采用小鼠肢体缺血模型，观察EPO对BMI的影响。EPO与BMI的同时施用通过细胞进一步分泌血管生成因子而增强植入的成红细胞的血管生成（JMCC 2006：40：629-38）。(2)事实如上所述（体外）：进行体外血管生成。预先从人类造血干细胞中产生的成红细胞集落在它们周围产生了毛细血管样的网络。EPO的加入进一步增强了血管生成，推测是通过对成红细胞的存活作用（JMCC 2006：40：629-38）。(3)成红细胞在血管生成中的重要作用：缺血肢体在移植了红细胞耗尽的骨髓细胞的小鼠中不能存活，因此成红细胞对于体内效应是必不可少的。移植纯化红系细胞的小鼠肢体未意外存活，同时移植CD 14阳性骨髓细胞的效果恢复。因此，BMI的血管生成作用是在巨噬细胞的辅助下由植入的红系细胞引起的（JMCC 2006：40：629-38）。少

A. Clinical trial of bone marrow implantation (BMI) accompanied with erythropoietin (EPO) injection(1) The results of monotherapy of BMI and combination therapy of BMI and EPO 6,000 IU : EPO injection did not cause major side effects, and no additional effects were observed in the combination therapy compared with the monotherapy. We reported that the number of implanted CD34-positive cells was one of the primary factors that influenced the clinical efficacy of BMI. (Circ J 2004;68:1189-93)(2) The results of combination therapy of BMI and high dose EPO 24,000IU : Four patients participated. A patient who had been previously treated with the monotherapy one year ago was treated again with the high dose combination therapy. The change of ankle-brachial pressure index revealed the superiority of the high dose combination : monotherapy, 0→0, and the high dose combination, 0→0.67. However, the other 3 patients showed any additional effect of the high dose EPO. Then we tried a combination th … More erapy of BMI and a sequential administration of EPO to obtain prolonged effects of EPO in the ischemic organs.(3) The results of the combination therapy of BMI and EPO 6,000 IU x 5-days : Four patients participated. One of them showed the improvement of rest pain, but the others did not, and suffered from leg amputation, later. Further device of combination therapy is necessary for such serious patients.B. Basic research of the mechanism of angiogenesis by erythroblasts and EPO(1) The additional effect of EPO to BMI (in vivo) : A murine model of limb ischemia was studied. Simultaneous administration of EPO with BMI enhanced angiogenesis by the implanted erythroblasts through further secretion of angiogenic factors by the cells (JMCC 2006:40:629-38).(2) The facts as stated above (in vitro) : In vitro angiogenesis was performed. Erythroblastic colonies made beforehand from human hematopoietic stem cells produced capillary-like networks around them. The addition of EPO further enhanced the angiogenesis presumably through surviving effects on erythroblasts (JMCC 2006:40:629-38).(3) The essential role of erythroblast in angiogenesis : Ischemic limbs did not survive in mice transplanted with erythroid-depleted bone marrow cells, hence erythroblasts were essential for the effect in vivo. Limbs did not survive unexpectedly in mice transplanted with purified erythroid cells, and the effects recovered by the simultaneous implantation of CD14-positive bone marrow cells. Therefore the angiogenic effects of BMI arose from the implanted erythroid cells in the presence of the assistance of macrophages (JMCC 2006:40:629-38). Less