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

骨髓移植联合促红细胞生成素注射的临床试验(1)单用促红细胞生成素及其与促红细胞生成素6,000 IU联合治疗的结果：促红细胞生成素注射无明显不良反应，联合治疗与单一治疗相比无其他副作用。我们报道，移植的CD34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联合大剂量EPO 24,000IU治疗结果：4例患者。一名一年前曾接受单一治疗的患者再次接受大剂量联合治疗。踝臂压指数的变化显示了大剂量联合用药的优越性：单一用药，0→0，高剂量联合用药，0→0.6 7。然而，其他3名患者表现出任何额外的影响，大剂量EPO。然后我们尝试了…的组合(3)BMI+EPO 6000IU×5天联合治疗4例。其中一例显示休息疼痛有所改善，但其他患者没有改善，后来进行了腿部截肢。B.红细胞和EPO促血管生成机制的基础研究(1)EPO对BMI(体内)的附加作用：建立小鼠肢体缺血模型。同时给予EPO和BMI通过进一步分泌血管生成因子来促进植入的红细胞的血管生成(JMCC 2006：40：629-38)。(2)上述事实(体外)：体外血管生成。由人类造血干细胞预先制成的红系集落在它们周围形成了毛细血管状的网络。EPO的加入进一步促进了血管的生成，可能是通过对红细胞的存活作用(JMCC 2006：40：629-38)。(3)红细胞在血管生成中的关键作用：在移植了红系耗尽的骨髓细胞的小鼠中，缺血的肢体无法存活，因此红细胞对于体内的作用是必不可少的。在移植了纯化的红系细胞的小鼠中，四肢没有意外地存活下来，通过同时移植CD14阳性的骨髓细胞，这种效果得到了恢复。因此，BMI的血管生成效应来自于在巨噬细胞的辅助下植入的红系细胞(JMCC 2006：40：629-38)。较少

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