Honold Joerg, Lehmann Ralf, Heeschen Christopher, Walter Dirk H, Assmus Birgit, Sasaki Ken-ichiro, Martin Hans, Haendeler Judith, Zeiher Andreas M, Dimmeler Stefanie
Department of Cardiology and Molecular Cardiology, Internal Medicine III, J.W. Goethe University of Frankfurt, Theodor-Stern-Kai 7, 60590 Frankfurt, Germany.
Arterioscler Thromb Vasc Biol. 2006 Oct;26(10):2238-43. doi: 10.1161/01.ATV.0000240248.55172.dd. Epub 2006 Aug 10.
Bone marrow-derived circulating endothelial progenitor cells (EPCs) may contribute to regeneration of infarcted myocardium and enhance neovascularization. Granulocyte colony-stimulating factor (G-CSF) is well-established to mobilize hematopoietic stem cells (HSCs) and might, thereby, also increase the pool of endogenously circulating EPC. Therefore, we investigated the effects of G-CSF administration on mobilization and functional activities of blood-derived EPC in patients with chronic ischemic heart disease (CIHD).
Sixteen patients with CIHD received 10 microg/kg per day subcutaneous G-CSF injection for 5 days. Leukocyte counts, the number of HSCs and EPCs, and the migratory response to VEGF and SDF-1 were analyzed before and after G-CSF-therapy. At day 5 of G-CSF treatment, the number of circulating leukocytes, CD34+ CD45+ and CD34+ CD133+ cells was significantly increased. Likewise, G-CSF treatment augmented the numbers of colony forming units with endothelial cell morphology (EC-CFU). However, the functional activity of the EPC as assessed by the migratory response to VEGF and SDF-1 was significantly reduced after G-CSF treatment (P<0.01). Because G-CSF was previously shown to cleave the CXCR4 receptor, we determined the surface expression of the 6H8 epitope of the CXCR4 receptor by fluorescence-activated cell sorter (FACS) analysis. Consistent with the reduced migratory capacity, the surface expression of the functionally active CXCR4 receptor was significantly reduced. To test the functional activity of the cultivated EPCs in vivo, cells were intravenously infused in nude mice after hind limb ischemia. EPCs, which were cultivated before G-CSF administration, increased blood flow recovery and prevented limb necrosis. However, infusion of EPCs, which were isolated 5 days after G-CSF treatment from the same patient, showed a reduced capacity to augment blood flow recovery and to prevent necrosis by 27%.
G-CSF treatment effectively mobilizes HSCs and EPCs. However, the migratory response to SDF-1 and in vivo capacity of G-CSF-mobilized EPCs was significantly reduced.
骨髓来源的循环内皮祖细胞（EPCs）可能有助于梗死心肌的再生并增强新生血管形成。粒细胞集落刺激因子（G-CSF）可有效动员造血干细胞（HSCs），因此也可能增加内源性循环EPC的数量。因此，我们研究了给予G-CSF对慢性缺血性心脏病（CIHD）患者血源性EPC动员和功能活性的影响。
16例CIHD患者接受每天10μg/kg皮下注射G-CSF，共5天。在G-CSF治疗前后分析白细胞计数、HSCs和EPCs数量以及对VEGF和SDF-1的迁移反应。在G-CSF治疗第5天时，循环白细胞、CD34+CD45+和CD34+CD133+细胞数量显著增加。同样，G-CSF治疗增加了具有内皮细胞形态的集落形成单位（EC-CFU）数量。然而，G-CSF治疗后，通过对VEGF和SDF-1的迁移反应评估的EPC功能活性显著降低（P<0.01）。由于先前已证明G-CSF可裂解CXCR4受体，我们通过荧光激活细胞分选仪（FACS）分析确定了CXCR4受体6H8表位的表面表达。与迁移能力降低一致，功能活性CXCR4受体的表面表达显著降低。为了在体内测试培养的EPCs的功能活性，在裸鼠后肢缺血后静脉注射细胞。在给予G-CSF之前培养的EPCs可增加血流恢复并预防肢体坏死。然而，注射在G-CSF治疗5天后从同一患者分离的EPCs，其增加血流恢复和预防坏死的能力降低了27%。
G-CSF治疗可有效动员HSCs和EPCs。然而，G-CSF动员的EPCs对SDF-1的迁移反应和体内能力显著降低。
