Abstract OBJECTIVE: To examine the number and function of circulating endothelial progenitor cells (EPCs) in children with cyanotic congenital heart diseases (CHD) and study their correlation with serum levels of vascular endothelial growth factor (VEGF) and stromal cell derived factor-1 (SDF-1). METHODS: Fifteen children with tetralogy of Fallot (cyanotic group) and 15 age-and sex-matched children with ventricular septal defect (control group) were enrolled. Serum levels of VEGF and SDF-1 were measured using ELISA. Mononuclear cells were isolated from peripheral blood by Ficoll density gradient centrifugation and cultured in vitro. EPCs were identified by immunofluorescence and were counted under a microscope. Modified Boyden chamber assay and the MTT assay were used to measure the migration and proliferation capacities of EPCs. EPCs adhesion ability assay was performed by replating cells on fibronectin-coated dishes, and then adherent cells were counted. The correlations of serum levels of VEGF and SDF-1 with the number and function of circulating EPCs were assessed by linear regression analysis. RESULTS: Serum levels of VEGF (201.42±44.74 ng/L vs 113.56±35.62 ng/L; P< 0.05) and SDF-1 (3.45±1.07 ng/L vs 1.05±0.99 ng/L; P<0.05) in the cyanotic group were higher than those in the control group. There was a positive correlation between serum levels of VEGF and SDF-1(r=0.675, P<0.01). The number of EPCs (×200 field) in the cyanotic group significantly increased compared with that of the control group (72.2±9.73 vs 51.2±3.83; P<0.01). The functional activities of EPCs, including proliferation, migration and adhesion capacities, were augmented in the cyanotic group compared with those in the control group. The increased number and function of EPCs and the increased serum levels of VEGF and SDF-1 were consistent in the cyanotic group, with a correlation coefficient of 0.8395, 0.5491, 0.6376 and 0.7392 respectively. CONCLUSIONS: The number and functional activity of EPCs as well as serum levels of VEGF and SDF-1 increased in children with cyanotic CHD. Serum levels of VEGF and SDF-1 were correlated to the number and functional activity of EPCs. Serum VEGF and SDF-1 together with circulating EPCs may play important roles in the pathology and physiology in these patients.［Chin J Contemp Pediatr, 2009, 11 (4):267-272］
LIU Zhe-Liang,WU Zhong-Shi,HU Jian-Guo et al. Correlation of serum levels of VEGF and SDF-1 with the number and function of circulating EPCs in children with cyanotic congenital heart disease[J]. 中国当代儿科杂志, 2009, 11(04): 267-272.
LIU Zhe-Liang,WU Zhong-Shi,HU Jian-Guo et al. Correlation of serum levels of VEGF and SDF-1 with the number and function of circulating EPCs in children with cyanotic congenital heart disease[J]. CJCP, 2009, 11(04): 267-272.
［1］Tepper OM, Sealove BA, Murayama T, Asahara T. Newly emerging concepts in blood vessel growth: recent discovery of endothelial progenitor cells and their function in tissue regeneration［J］. J Investig Med, 2003, 51(6):353-359.
［2］Wilhelm C, Bal L, Smirnov P, Galy-Fauroux I, Clément O, Gazeau F, et al. Magnetic control of vascular network formation with magnetically labeled endothelial progenitor cells［J］. Biomaterials, 2007, 28(26):3797-3806.
［3］Rodriguez-Losada N, Garcia-Pinilla JM, Jimenez-Navarro MF, Gonzalez FJ. Endothelial progenitor cells in cell-based therapy for cardiovascular disease［J］. Cell Mol Biol, 2008, 54(1):11-23.
［4］Khoo CP, Pozzilli P, Alison MR. Endothelial progenitor cells and their potential therapeutic applications［J］. Regen Med, 2008, 3(6):863-876.
［5］Smythe J, Fox A, Fisher N, Frith E, Harris AL, Watt SM. Measuring angiogenic cytokines, circulating endothelial cells, and endothelial progenitor cells in peripheral blood and cord blood: VEGF and CXCL12 correlate with the number of circulating endothelial progenitor cells in peripheral blood［J］. Tissue Eng Part C Methods, 2008, 14(1):59-67.
［6］Hoenig MR, Bianchi C, Rosenzweig A, Sellke FW. Decreased vascular repair and neovascularization with ageing: mechanisms and clinical relevance with an emphasis on hypoxia-inducible factor-1［J］. Curr Mol Med, 2008, 8(8):754-767.
［7］Yamashita J, Itoh H, Hirashima M, Ogawa M, Nishikawa S, Yurugi T, et al. Flk1-positive cells derived from embryonic stem cells serve as vascular progenitors［J］.Nature, 2000, 408(6808):92-96.
［8］Urbich C, Dimmeler S. Endothelial progenitor cells: characterization and role in vascular biology［J］.Circ Res, 2004, 95（4）: 343-353.
［9］Hiasa K, Ishibashi M, Ohtani K, Inoue S, Zhao Q, Kitamoto S, et al. Gene transfer of stromal cell-derived factor-1 enhances ischemic vasculogenesis and angiogenesis via vascular endothelial growth factor/ endothelial nitric oxide synthase related pathway: next-generation chemokine therapy for therapeutic neovascularization［J］.Circulation, 2004, 109(20):2454-2461.
［10］Rodriguez-Losada N, Garcia-Pinilla JM, Jimenez-Navarro MF, Gonzalez FJ. Endothelial progenitor cells in cell-based therapy for cardiovascular disease［J］. Cell Mol Biol, 2008, 54(1):11-23.
［11］Asahara T, Murohara T, Sullivan A, Silver M, van der Zee R, Li T, et al. Isolation of putative progenitor endothelial cells for angiogenesis［J］.Science, 1997, 275（5302）:964-967.
［12］Peichev M, Naiyer AJ, Pereira D, Zhu Z, Lane WJ, Williams M, et al. Expression of VEGFR-2 and AC133 by circulating human CD34(+) cells identifies a population of functional endothelial precursors［J］.Blood, 2000, 95(3):952-958.
［13］Rafii S, Lyden D. Therapeutic stem and progenitor cell transplantation for organ vascularization and regeneration［J］.Nat Med, 2003, 9(6):702-712.
［14］Yamaguchi J, Kusano KF, Masuo O, Kawamoto A, Silver M, Murasawa S, et al. Stromal cell derived factor-1 effects on exvivo expanded endothelial progenitor cell recruitment for ischemic neovascularization［J］. Circulation, 2003, 107(9):1322-1328.
［15］Ceradini DJ, Kulkarni AR, Callaghan MJ, Tepper OM, Bastidas N, Kleinman ME,et al. Progenitor cell trafficking is regulated by hypoxic gradients through HIF-1 induction of SDF-1［J］.Nat Med, 2004, 10(8):858-864.
［16］Manalo DJ, Rowan A, Lavoie T, Natarajan L, Kelly BD, Ye SQ, et al. Transcriptional regulation of vascular endothelial cell responses to hypoxia by HIF-1［J］.Blood, 2005, 105(2):659-669.
［17］Heissig B, Hattori K, Dias S, Friedrich M, Ferris B, Hackett NR, et al. Recruitment of stem and progenitor cells from the bone marrow niche Mmp-9 mediated release of Kit-ligand［J］.Cell, 2002, 109 (5):625-637.
［18］Hitchon C, Wong K, Ma G, Reed J, Lyttle D, El-Gabalawy H. Hypoxia-induced production of stromal cell-derived factor 1（CXCL12）and vascular endothelial growth factor by synovial fibroblasts［J］.Arthritis Rheum, 2002, 46(10):2587-2597.
［19］Dar A, Goichberg P, Shinder V, Kalinkovich A, Kollet O, Netzer N, et al. Chemokine receptor CXCR4-dependent internalization and resecretion of functional chemokine SDF-1 by bone marrow endothelial and stromal cells［J］.Nat Immunol, 2005, 6（10）:1038-1046.
［20］Badorff C, Brandes RP, Popp R, Rupp S, Urbich C, Aicher A，et al. Transdifferentiation of blood-derived human adult endothelial progenitor cells into functionally active cardiomyocytes［J］.Circulation, 2003,107(7):1024-1032.
［21］Kortesidis A, Zannettino A, Isenmann S, Shi S, Lapidot T, Gronthos S. Stromal-derived factor-1 promotes the growth, survival, and development of human bone marrow stromal stem cells［J］. Blood, 2005, 105(10):3793-3801.
［22］Wu H, Riha GM, Yang H, Li M, Yao Q, Chen C. Differentiation and proliferation of endothelial progenitor cells from canine peripheral blood mononuclear cells［J］. J Surg Res, 2005, 126(2):193-198.
