摘要
研究背景脉络膜新生血管(choroidal neovascularization, CNV)是年龄相关性黄斑变性(age related macular degeneration, AMD)导致严重视力丧失最主要的原因;其发病是一多因素参与的复杂过程,但确切机制仍未阐明。血管生成和血管发生是参与新生血管形成的两种机制。一般来说,局部促血管生成因子参与血管生成,其主要细胞成份来源于已存在的毛细血管。然而,近年研究发现外周血骨髓来源的内皮祖细胞(endothelial precursor cells, EPCs)可能参与病理性新生血管形成,包括视网膜和脉络膜新生血管。作为CXC趋化蛋白超家族成员之一,基质细胞衍生因子-1(stromal cell derived factor-1, SDF-1)参与趋化骨髓来源细胞(bone marrow derived cells, BMCs)向靶组织募集,其受体CXCR4可表达于多种细胞,包括内皮细胞及EPCs。
体内、外研究证明,SDF-1也是一种促血管生成因素。SDF-1或CXCR4基因缺陷小鼠中消化道大血管发育缺失。增殖性视网膜病变和CNV模型显示,SDF-1/CXCR4可诱导EPCs聚集于新生血管形成部位,而CXCR4拮抗剂可明显抑制包括缺血性视网膜新生血管和CNV在内的多种眼内新生血管病变。临床研究表明,正常老年人及AMD患者脉络膜中,视网膜色素上皮(retinal pigment epithelium, RPE)和脉络膜损伤处SDF-1和CXCR4高表达。组织缺血时,缺氧诱导因子-1α(hypoxia inducible factor-1α, HIF-1α)可上调SDF-1表达,介导EPCs向缺血组织聚集,而CNV形成过程中也存在HIF-1α表达增加。RPE细胞可持续表达SDF-1,并在缺氧时表达增加。然而,SDF-1在CNV发生中的作用机制尚未完全阐明。
目的和内容探讨SDF-1在CNV形成过程中的作用,进一步阐明CNV的发生机制。在激光诱导的小鼠CNV模型中,观察HIF-1α和SDF-1表达;体外通过化学缺氧及运用HIF-1小干扰RNA(small interfering RNA, siRNA)观察SDF-1表达。观察C57BL/6J-GFP嵌合体小鼠CNV模型中SDF-1诱导骨髓来源的EPCs和内皮细胞(endothelial cells, ECs)在CNV局部的聚集;体外模拟在不同SDF-1表达水平及微环境下新生血管生成情况,观察SDF-1对BMCs分化的影响。
方法1)利用532nm激光建立小鼠CNV模型,观察CNV局部HIF-1α和SDF-1的表达;利用体外培养RPE细胞,观察缺氧条件下,RPE细胞HIF-1α和SDF-1表达的情况;运用siRNA技术构建HIF-1α低表达的RPE细胞,观察缺氧状态下HIF-1α缺失对于RPE细胞SDF-1表达的影响;2)建立C57BL/6J-GFP嵌合体小鼠CNV模型,观察CNV局部SDF-1表达及BMCs表面CXCR4、CD34、CD31和c-kit表达;3)体外培养人原代BMCs和RF/6A细胞,通过三维共培养技术,比较不同SDF-1表达水平及不同微环境下,BMCs参与新生血管形成及发生分化的情况。
结果免疫荧光染色显示CNV部位HIF-1α表达升高,于24h达峰值;随之SDF-1表达也升高,于3d达峰值,表达部位均位于近RPE层附近。缺氧诱导体外培养RPE细胞HIF-1α及SDF-1表达增高,于4小时达峰值;shHIF-1α抑制HIF-1α表达,同时SDF-1表达下调。脉络膜铺片及冰冻切片间接免疫荧光染色结果表明,在CNV形成极早期,GFP阳性的BMCs主要聚集于SDF-1高表达的光斑周围的RPE层附近,形态较均一,其中包含大量EPCs细胞;CNV形成过程中,大量BMCs细胞进入损伤内部,部分细胞胞体变狭长,分化为内皮细胞(CD31+)形成管腔样结构,此时损伤周边仍聚集大量的EPCs;随CNV形成,可见大量新生血管管腔样结构交织成网,大量BMCs细胞分化的内皮细胞胞参与其形成。CNV损伤局部聚集的BMCs均为CXCR4阳性,其中CD34阳性和/或c-kit阳性EPCs位于损伤周边部,而同来源的CD31阳性内皮细胞随CNV发展,渐伸入损伤核心区,参与新生血管形成。体外三维胶原纤维基质模型显示,SDF-1高表达组新生血管密集,单纯SDF-1升高即可诱导BMCs向内皮细胞分化,参与新生血管形成。
结论在CNV形成过程中,缺氧诱导RPE层HIF-1α表达增高,上调SDF-1表达;SDF-1/CXCR4趋化BMCs聚集于CNV局部,并诱导部分BMCs向内皮细胞分化,参与CNV新生血管形成。
Background Choroidal neovascularization (CNV) is a major cause of vision loss in patients with age related macular degeneration (AMD). The pathogenesis of CNV is clearly multifactorial, but not completely understood. Generally, neovascularization incorporates two forms of new blood vessel growth: angiogenesis and vasculogenesis. Angiogenesis were generally promoted by growth factors, and most cellular components of the new vessel complex (endothelial cells, smooth muscle cells, and pericytes) were derived from cells resident within adfacent pre-existing blood vessels. However, more and more studies in animal models suggest that circulating bone marrow-derived endothelial precursor cells (EPCs) may be participate in pathophysiological noevascularization, including retinal and choroidal neovascularization.
As a member of the CXC chemokine subfamily, stromal cell derived factor 1 (SDF-1) is implicated for CXCR4-positive EPCs recruitment from the bone marrow to the target tissue. CXCR4, a seven transmembrane spanning G protein coupled receptor, is the only known receptor for SDF-1 and is expressed on various endothelial cells and EPCs. It has been reported that SDF-1 acts as an angiogenic agent in several in vivo and in vitro model systems. Mice deficient in either SDF-1 or CXCR4 had defective formation of large vessels supplying the gastrointestinal tract. In proliferative retinopathy and CNV model, SDF-1/CXCR4 may have a role in orienting EPC to reach sites of neovascularization. Besides, CXCR4 antagonists were effective supppressors of several types of ocular NV, such as ischemia-induced retinal NV and CNV. Clinically, in aged control and AMD choroid, SDF-1 and CXCR4 were most prominent in retinal pigment epithelial (RPE) cells and choroidal stroma.
Some studies showed that SDF-1 was expressed in ischemic tissues under the control of HIF-1α. In models of wound healing and several models of tissue hypoxia, SDF-1 mediated homing of endothelial progenitor cells to blood vessel walls in the ischemic tissue. It was concluded that recruitment of CXCR4 positive progenitor cells to regenerating tissues were mediated by hypoxic gradients via HIF-1α-induced expression of SDF-1. HIF-1αinduced SDF-1 expression facilitated adhesion of progenitor cells to ischemic endothelium and induced their migration in transwell assays, suggesting a specific role in homing of circulating progenitor cells to ischemic vascular beds. In this study, we will investigate the potential role of hypoxia induced SDF-1 expression in formation of CNV.
Objectives It would be helpful for our understanding of CNV to study the effect of SDF-1 on CNV formation. In this study, we investigated the expression of HIF-1αand SDF-1 in CNV models; then, we studied the SDF-1 expression in cultured RPE treated with CoCl2 and HIF-1siRNA. Then, we used C57BL/6J-GFP chimeric mice to investigate the SDF-1 induced bone marrow derived-EPCs to CNV. Finally, We established an in vitro 3-dimensional (3D) matrix system to study the neovascularization under different SDF-1 levels and microenviroments.
Methods 1) The C57BL/6J mice underwent laser rupture of Bruch’s membrane to induce CNV and were killed at 1, 4, 8, 16, 24 hours and 1, 3, 7, 14, 28 days after laser injury. Immunofluorescence analysis were processed the expression of HIF-1αand SDF-1. Cultured human RPE cells were exposed to hypoxia (CoCl2, 0.2mM) for 0, 1, 2, 4, 8, 16, 24 and 48 hours, HIF-1αand SDF-1 was detected with Western Blot, RT-PCR and ELISA respetively. HIF-1αshRNA was used to knock down the HIF-1αgene in hRPE, the expression of HIF-1αand SDF-1 under hypoxia was detected. 2) Green fluorescent protein (GFP) chimeric mice were developed by transplanting bone marrow cells from GFP+/+ transgenic mice to adult C57BL/6J mice and established CNV model after 1 month when confirmed the successful bone marrow transplantation. The chimeric mice were killed at 1, 3, 7, 14 and 28 days after laser injury. The eyes were enucleated and frozen section. SDF-1, CXCR4, CD34, CD31 and c-kit were detected by immunofluorescence. The recruitment of BMCs to CNV is measured by choroidal flatmount. 3) 3D fibrin matrix system was prepared in vitro with human fibrinogen, thrombin and aprotinin. Cultured human BMCs (marked with DiO) and RF/6A cells were seed in the gel, cocultured with different conditional media. The tubular structure and BMCs participating in neovascularization were investigated with a phase contrast microscope. Gels were fixed in 4% buffered formalin and processed for paraffin embedding. The embedded matrixes were cut (5μm) perpendicularly to the surface of the matrix sheet. The expression of CD34 and CD31 on BMCs was examined by immunofluorescence.
Results Immunofluorescence showed that the expression of HIF-1αand SDF-1 in CNV were near RPE layer, which were gradually higher and reached the peak at 24 hour and 3d respectively after laser injury. RT-PCR and Western Blot showed that the hypoxic stimulus induced the expression of HIF-1αand SDF-1 in cultured hRPE, and reached the peak at 4th hour, while shHIF-1αhad knockdown the expression of HIF-1αand down-regulated the SDF-1 level. Choroidal flatmount showed that at the early stage of CNV, BMCs gathered around RPE layer near the laser lesion; the GFP-labeled cells were morphologically green fluorescent round spots. During CNV formation, CXCR4 marked BMCs in CNV lesion were differentiated into endothelial cells (CD31+) and participated in vessel formation. When CNV came into being, there were amount of BMCs derived endothelial cells participating in the neovasculature network. Immunofluorescence showed that most BMCs gathered in CNV were CXCR4 positive cells, and some acquired phenotypic markers of EPCs (CD34 and c-kit) and ECs (CD31). In the RF/6A-BMCs-cocultured 3D fibrin matrix system, the DiO labled BMCs were particitated in the tubular networks. There were more CD31 positive BMCs (Dio labeled) in SDF-1 constitutive-expression, SDF-1 and hypoxia group.
Conclusion During CNV formation, the hypoxia induced higher expression of HIF-1αin RPE layer, which up-regulated the SDF-1 expression. SDF-1 and its receptor CXCR4 were critical mediators for ischemia-specific recruitment to CNV. Also did SDF-1 can induce BMCs differentiate into endothelial cells to participate in the neovascularization.
引文
1. Zampetaki A, Kirton JP, Xu Q. Vascular repair by endothelial progenitor cells. Cardiovasc Res 2008;78:413-421.
2. Chan-Ling T, Baxter L, Afzal A, et al. Hematopoietic Stem Cells Provide Repair Functions after Laser-Induced Bruch's Membrane Rupture Model of Choroidal Neovascularization. Am J Pathol 2006;168:1031-1044.
3. Sengupta N, Caballero S, Mames RN, Butler JM, Scott EW, Grant MB. The Role of Adult Bone Marrow-Derived Stem Cells in Choroidal Neovascularization. Invest Ophthalmol Vis Sci 2003;44:4908-4913.
4. Espinosa-Heidmann DG, Caicedo A, Hernandez EP, Csaky KG, Cousins SW. Bone Marrow-Derived Progenitor Cells Contribute to Experimental Choroidal Neovascularization. Invest Ophthalmol Vis Sci 2003;44:4914-4919.
5. Espinosa DG, Caicedo A, Hernandez EP, Cousins SW. Adult Bone Marrow Derived Progenitor Cells Contribute to Choroidal Neovascularization and Modulate the Severity. Invest Ophthalmol Vis Sci 2003;44:3936-.
6. Espinosa DG, Caicedo A, Hernandez EP, et al. Aged bone marrow transfers age-related pathology into young recipients in experimental choroidal neovascularization (CNV). Invest Ophthalmol Vis Sci 2004;45:1853-.
7. Harris JR, Brown GAJ, Jorgensen M, et al. Bone Marrow-Derived Cells Home to and Regenerate Retinal Pigment Epithelium after Injury. Invest Ophthalmol Vis Sci 2006;47:2108-2113.
8. Hirota K, Semenza GL. Regulation of angiogenesis by hypoxia-inducible factor 1. Critical Reviews in Oncology/Hematology 2006;59:15-26.
9. Tong J-P, Yao Y-F. Contribution of VEGF and PEDF to choroidal angiogenesis: A need for balanced expressions. Clinical Biochemistry 2006;39:267-276.
10. Penfold PL, Madigan MC, Gillies MC, Provis JM. Immunological and aetiological aspects of macular degeneration. Prog Retin Eye Res 2001;20:385-414.
11. Ferrara N, Damico L, Shams N, Lowman H, Kim R. Development of ranibizumab, an anti-vascular endothelial growth factor antigen binding fragment, as therapy for neovascular age-related macular degeneration. Retina 2006;26:859-870.
12. Kinose F, Roscilli G, Lamartina S, et al. Inhibition of retinal and choroidal neovascularization by a novel KDR kinase inhibitor. Mol Vis 2005;11:366-373.
13. Yuji Oshima SOHNSKSFHMMMKSCNEDJZPAC. Increased expression of VEGF in retinal pigmented epithelial cells is not sufficient to cause choroidal neovascularization. Journal of Cellular Physiology 2004;201:393-400.
14. Ceradini DJ, Gurtner GC. Homing to hypoxia: HIF-1 as a mediator of progenitor cellrecruitment to injured tissue. Trends Cardiovasc Med 2005;15:57-63.
15. Ceradini DJ, Kulkarni AR, Callaghan MJ, et al. Progenitor cell trafficking is regulated by hypoxic gradients through HIF-1 induction of SDF-1. Nat Med 2004;10:858-864.
16. De Falco E, Porcelli D, Torella AR, et al. SDF-1 involvement in endothelial phenotype and ischemia-induced recruitment of bone marrow progenitor cells. Blood 2004;104:3472-3482.
17. Guleng B, Tateishi K, Ohta M, et al. Blockade of the Stromal Cell-Derived Factor-1/CXCR4 Axis Attenuates In vivo Tumor Growth by Inhibiting Angiogenesis in a Vascular Endothelial Growth Factor-Independent Manner. Cancer Res 2005;65:5864-5871.
18. Grant MB, Segal MS, Afzal A, et al. Stromal Derived Factor-1 Affects Multiple Steps in Stem Cell Recruitment to Areas of Ocular Neovascularization. Invest Ophthalmol Vis Sci 2004;45:455-.
19. Butler JM, Guthrie SM, Koc M, Afzal A, et al. SDF-1 is both necessary and sufficient to promote proliferative retinopathy. Journal of Clinical Investigation 2005;115:86-93.
20. Zhao W, Wang YS, Hui YN, et al. Inhibition of proliferation, migration and tube formation of choroidal microvascular endothelial cells by targeting HIF-1alpha with short hairpin RNA-expressing plasmid DNA in human RPE cells in a coculture system. Graefes Arch Clin Exp Ophthalmol 2008;246:1413-1422.
21. Bhutto IA, McLeod DS, Merges C, Hasegawa T, Lutty GA. Localisation of SDF-1 and its receptor CXCR4 in retina and choroid of aged human eyes and in eyes with age related macular degeneration. Br J Ophthalmol 2006;90:906-910.
22. Guerin EP, Sheridan C, Kent D, Grant MB, Hiscott P. Stromal Cell Derived Factor (SDF) 1 Alpha / CXCR4 Axis Components in Excised Choroidal Neovascular Membranes. Invest Ophthalmol Vis Sci 2006;47:1427-.
23. Crane IJ, Wallace CA, McKillop-Smith S, Forrester JV. CXCR4 receptor expression on human retinal pigment epithelial cells from the blood-retina barrier leads to chemokine secretion and migration in response to stromal cell-derived factor 1 alpha. J Immunol 2000;165:4372-4378.
24. Friedman DS, O'Colmain BJ, Munoz B, et al. Prevalence of age-related macular degeneration in the United States. Arch Ophthalmol 2004;122:564-572.
25. Cohen SY, Laroche A, Leguen Y, Soubrane G, Coscas GJ. Etiology of choroidal neovascularization in young patients. Ophthalmology 1996;103:1241-1244.
26. Lubin JR, Gragoudas ES, Albert DM. Choroidal neovascularization associated with malignant melanoma: a case report. Acta Ophthalmol (Copenh) 1982;60:412-418.
27. Guerin E, Hiscott P, Damato B. Choroidal neovascular membrane in a series of cases of malignant melanoma of the choroid. Acta Ophthalmol Scand 2006;84:323-327.
28.王雨生,朱洁.脉络膜新生血管性疾病的治疗现状.第四军医大学学报2006;27:481-483.
29.侯慧媛,王雨生.骨髓来源细胞与脉络膜新生血管.中华眼底病杂志2006;22:280-282.
30. Vinores SA, Xiao WH, Aslam S, et al. Implication of the hypoxia response element of theVegf promoter in mouse models of retinal and choroidal neovascularization, but not retinal vascular development. J Cell Physiol 2006;206:749-758.
31. Zhang P, Wang Y, Hui Y, et al. Inhibition of VEGF expression by targeting HIF-1 alpha with small interference RNA in human RPE cells. Ophthalmologica 2007;221:411-417.
32. Yang XM, Wang YS, Zhang J, et al. Role of PI3K/Akt and MEK/ERK in Mediating Hypoxia-Induced Expression of Hypoxia-Inducible Factor 1alpha and Vascular Endothelial Growth Factor in Laser-Induced Rat Choroidal Neovascularization. Invest Ophthalmol Vis Sci 2008.
33. Tomita M, Yamada H, Adachi Y, et al. Choroidal Neovascularization Is Provided by Bone Marrow Cells. Stem Cells 2004;22:21-26.
34. Zhang SX, Ma J-x. Ocular neovascularization: Implication of endogenous angiogenic inhibitors and potential therapy. Progress in Retinal and Eye Research 2007;26:1-37.
35. Patan S. Vasculogenesis and angiogenesis. Cancer Treat Res 2004;117:3-32.
36. Hughes S, Yang H, Chan-Ling T. Vascularization of the human fetal retina: roles of vasculogenesis and angiogenesis. Invest Ophthalmol Vis Sci 2000;41:1217-1228.
37. Chan-Ling T, McLeod DS, Hughes S, et al. Astrocyte-endothelial cell relationships during human retinal vascular development. Invest Ophthalmol Vis Sci 2004;45:2020-2032.
38. Csaky KG, Baffi JZ, Byrnes GA, et al. Recruitment of marrow-derived endothelial cells to experimental choroidal neovascularization by local expression of vascular endothelial growth factor. Exp Eye Res 2004;78:1107-1116.
39. Rafii S, Lyden D. Therapeutic stem and progenitor cell transplantation for organ vascularization and regeneration. Nat Med 2003;9:702-712.
40. Asahara T, Murohara T, Sullivan A, et al. Isolation of putative progenitor endothelial cells for angiogenesis. Science 1997;275:964-967.
41. Lin Y, Weisdorf DJ, Solovey A, Hebbel RP. Origins of circulating endothelial cells and endothelial outgrowth from blood. J Clin Invest 2000;105:71-77.
42. Gehling UM, Ergun S, Schumacher U, et al. In vitro differentiation of endothelial cells from AC133-positive progenitor cells. Blood 2000;95:3106-3112.
43. Yoder MC, Mead LE, Prater D, et al. Redefining endothelial progenitor cells via clonal analysis and hematopoietic stem/progenitor cell principals. Blood 2007;109:1801-1809.
44. Lopez PF, Grossniklaus HE, Lambert HM, et al. Pathologic features of surgically excised subretinal neovascular membranes in age-related macular degeneration. Am J Ophthalmol 1991;112:647-656.
45. Takahashi T, Kalka C, Masuda H, et al. Ischemia- and cytokine-induced mobilization of bone marrow-derived endothelial progenitor cells for neovascularization. Nat Med 1999;5:434-438.
46. Hou HY, Wang YS, Xu JF, Wang YC, Liu JP. The dynamic conduct of bone marrow-derived cells in the choroidal neovascularization microenvironment. Curr Eye Res 2006;31:1051-1061.
47. Persons DA, Allay JA, Riberdy JM, et al. Use of the green fluorescent protein as a marker to identify and track genetically modified hematopoietic cells. Nat Med 1998;4:1201-1205.
48. Gill M, Dias S, Hattori K, et al. Vascular trauma induces rapid but transient mobilization of VEGFR2(+)AC133(+) endothelial precursor cells. Circ Res 2001;88:167-174.
49.杨秀梅,王雨生,惠延年.外源性脉络膜新生血管抑制剂的研究进展. 2005 2005;25:75-78.
50. Asahara T, Takahashi T, Masuda H, et al. VEGF contributes to postnatal neovascularization by mobilizing bone marrow-derived endothelial progenitor cells. EMBO J 1999;18:3964-3972.
51. Shalaby F, Rossant J, Yamaguchi TP, et al. Failure of blood-island formation and vasculogenesis in Flk-1-deficient mice. Nature 1995;376:62-66.
52. Reinoso MA, Caicedo A, Espinosa-Heidmann DG, Csaky KH, Cousins SW. Blood-derived macrophages and Bone Marrow Derived Progenitor Cells contribute to intrachoroidal vascular changes in experimental Choroidal Neovascularization (CNV). Invest Ophthalmol Vis Sci 2004;45:3386-.
53. Hattori K, Heissig B, Tashiro K, et al. Plasma elevation of stromal cell-derived factor-1 induces mobilization of mature and immature hematopoietic progenitor and stem cells. Blood 2001;97:3354-3360.
54. Togel F, Isaac J, Hu Z, Weiss K, Westenfelder C. Renal SDF-1 signals mobilization and homing of CXCR4-positive cells to the kidney after ischemic injury. Kidney Int 2005;67:1772-1784.
55. Askari AT, Unzek S, Popovic ZB, et al. Effect of stromal-cell-derived factor 1 on stem-cell homing and tissue regeneration in ischaemic cardiomyopathy. Lancet 2003;362:697-703.
56. Nishikawa S, Ogawa M, Kunisada T, Kodama H. B lymphopoiesis on stromal cell clone: stromal cell clones acting on different stages of B cell differentiation. Eur J Immunol 1988;18:1767-1771.
57. Nagasawa T, Hirota S, Tachibana K, et al. Defects of B-cell lymphopoiesis and bone-marrow myelopoiesis in mice lacking the CXC chemokine PBSF/SDF-1. Nature 1996;382:635-638.
58. Pablos JL, Amara A, Bouloc A, et al. Stromal-cell derived factor is expressed by dendritic cells and endothelium in human skin. Am J Pathol 1999;155:1577-1586.
59. Kollet O, Shivtiel S, Chen YQ, et al. HGF, SDF-1, and MMP-9 are involved in stress-induced human CD34+ stem cell recruitment to the liver. J Clin Invest 2003;112:160-169.
60. Stumm RK, Rummel J, Junker V, et al. A dual role for the SDF-1/CXCR4 chemokine receptor system in adult brain: isoform-selective regulation of SDF-1 expression modulates CXCR4-dependent neuronal plasticity and cerebral leukocyte recruitment after focal ischemia. J Neurosci 2002;22:5865-5878.
61. Tachibana K, Hirota S, Iizasa H, et al. The chemokine receptor CXCR4 is essential for vascularization of the gastrointestinal tract. Nature 1998;393:591-594.
62. Wojakowski W, Tendera M, Michalowska A, et al. Mobilization of CD34/CXCR4+,CD34/CD117+, c-met+ stem cells, and mononuclear cells expressing early cardiac, muscle, and endothelial markers into peripheral blood in patients with acute myocardial infarction. Circulation 2004;110:3213-3220.
63. Ratajczak MZ, Majka M, Kucia M, et al. Expression of functional CXCR4 by muscle satellite cells and secretion of SDF-1 by muscle-derived fibroblasts is associated with the presence of both muscle progenitors in bone marrow and hematopoietic stem/progenitor cells in muscles. Stem Cells 2003;21:363-371.
64. Schrader AJ, Lechner O, Templin M, et al. CXCR4/CXCL12 expression and signalling in kidney cancer. Br J Cancer 2002;86:1250-1256.
65. Ponomaryov T, Peled A, Petit I, et al. Induction of the chemokine stromal-derived factor-1 following DNA damage improves human stem cell function. J Clin Invest 2000;106:1331-1339.
66. Wistow G, Bernstein SL, Ray S, et al. Expressed sequence tag analysis of adult human iris for the NEIBank Project: steroid-response factors and similarities with retinal pigment epithelium. Mol Vis 2002;8:185-195.
67. Bonaldo MF, Lennon G, Soares MB. Normalization and subtraction: two approaches to facilitate gene discovery. Genome Res 1996;6:791-806.
68. Bleul CC, Farzan M, Choe H, et al. The lymphocyte chemoattractant SDF-1 is a ligand for LESTR/fusin and blocks HIV-1 entry. Nature 1996;382:829-833.
69. Oberlin E, Amara A, Bachelerie F, et al. The CXC chemokine SDF-1 is the ligand for LESTR/fusin and prevents infection by T-cell-line-adapted HIV-1. Nature 1996;382:833-835.
70. Balabanian K, Lagane B, Infantino S, et al. The Chemokine SDF-1/CXCL12 Binds to and Signals through the Orphan Receptor RDC1 in T Lymphocytes. J Biol Chem 2005;280:35760-35766.
71. Burns JM, Summers BC, Wang Y, et al. A novel chemokine receptor for SDF-1 and I-TAC involved in cell survival, cell adhesion, and tumor development. J Exp Med 2006;203:2201-2213.
72. Proost P, Mortier A, Loos T, et al. Proteolytic processing of CXCL11 by CD13/aminopeptidase N impairs CXCR3 and CXCR7 binding and signaling and reduces lymphocyte and endothelial cell migration. Blood 2007;110:37-44.
73. Dar A, Goichberg P, Shinder V, et al. Chemokine receptor CXCR4-dependent internalization and resecretion of functional chemokine SDF-1 by bone marrow endothelial and stromal cells. Nat Immunol 2005;6:1038-1046.
74. Heissig B, Hattori K, Dias S, et al. Recruitment of stem and progenitor cells from the bone marrow niche requires MMP-9 mediated release of kit-ligand. Cell 2002;109:625-637.
75. Aicher A, Heeschen C, Mildner-Rihm C, et al. Essential role of endothelial nitric oxide synthase for mobilization of stem and progenitor cells. Nat Med 2003;9:1370-1376.
76. Kollet O, Dar A, Shivtiel S, et al. Osteoclasts degrade endosteal components and promote mobilization of hematopoietic progenitor cells. Nat Med 2006;12:657-664.
77. Urbich C, Heeschen C, Aicher A, et al. Cathepsin L is required for endothelial progenitor-91-cell-induced neovascularization. Nat Med 2005;11:206-213.
78. Agnihotri AK, Recanati MA, White JK, et al. Continuous addition of adenosine with a micropump system improves warm whole blood cardioplegia. Heart Surg Forum 2003;6:264-272.
79. Avigdor A, Goichberg P, Shivtiel S, et al. CD44 and hyaluronic acid cooperate with SDF-1 in the trafficking of human CD34+ stem/progenitor cells to bone marrow. Blood 2004;103:2981-2989.
80. Wysoczynski M, Reca R, Ratajczak J, et al. Incorporation of CXCR4 into membrane lipid rafts primes homing-related responses of hematopoietic stem/progenitor cells to an SDF-1 gradient. Blood 2005;105:40-48.
81. Vajkoczy P, Blum S, Lamparter M, et al. Multistep nature of microvascular recruitment of ex vivo-expanded embryonic endothelial progenitor cells during tumor angiogenesis. J Exp Med 2003;197:1755-1765.
82. Yao L, Salvucci O, Cardones AR, et al. Selective expression of stromal-derived factor-1 in the capillary vascular endothelium plays a role in Kaposi sarcoma pathogenesis. Blood 2003;102:3900-3905.
83. Chavakis E, Aicher A, Heeschen C, et al. Role of beta2-integrins for homing and neovascularization capacity of endothelial progenitor cells. J Exp Med 2005;201:63-72.
84. Wu Y, Ip JE, Huang J, et al. Essential role of ICAM-1/CD18 in mediating EPC recruitment, angiogenesis, and repair to the infarcted myocardium. Circ Res 2006;99:315-322.
85. Duan H, Cheng L, Sun X, et al. LFA-1 and VLA-4 involved in human high proliferative potential-endothelial progenitor cells homing to ischemic tissue. Thromb Haemost 2006;96:807-815.
86. Massberg S, Konrad I, Schurzinger K, et al. Platelets secrete stromal cell-derived factor 1alpha and recruit bone marrow-derived progenitor cells to arterial thrombi in vivo. J Exp Med 2006;203:1221-1233.
87. Jin DK, Shido K, Kopp H-G, et al. Cytokine-mediated deployment of SDF-1 induces revascularization through recruitment of CXCR4+ hemangiocytes. Nat Med 2006;12:557-567.
88. Yamaguchi J-i, Kusano KF, Masuo O, et al. Stromal Cell-Derived Factor-1 Effects on Ex Vivo Expanded Endothelial Progenitor Cell Recruitment for Ischemic Neovascularization. Circulation 2003;107:1322-1328.
89. Li Y, Atmaca-Sonmez P, Schanie CL, Ildstad ST, Kaplan HJ, Enzmann V. Endogenous Bone Marrow Derived Cells Express Retinal Pigment Epithelium Cell Markers and Migrate to Focal Areas of RPE Damage. Invest Ophthalmol Vis Sci 2007;48:4321-4327.
90. Stellos K, Langer H, Daub K, et al. Platelet-Derived Stromal Cell-Derived Factor-1 Regulates Adhesion and Promotes Differentiation of Human CD34+ Cells to Endothelial Progenitor Cells. Circulation 2008;117:206-215.
91. De Falco E, Avitabile D, Totta P, et al. Altered Sdf-1-Mediated Diffferentiation of Bone Marrow-Derived Endothelial Progenitor Cells in Diabetes Mellitus. J Cell Mol Med 2009.
92. Salvucci O, Yao L, Villalba S, Sajewicz A, Pittaluga S, Tosato G. Regulation of endothelial cell branching morphogenesis by endogenous chemokine stromal-derived factor-1. Blood 2002;99:2703-2711.
93. Kanda S, Mochizuki Y, Kanetake H. Stromal Cell-derived Factor-1alpha Induces Tube-like Structure Formation of Endothelial Cells through Phosphoinositide 3-Kinase. J Biol Chem 2003;278:257-262.
94. Salcedo R, Wasserman K, Young HA, et al. Vascular Endothelial Growth Factor and Basic Fibroblast Growth Factor Induce Expression of CXCR4 on Human Endothelial Cells : In Vivo Neovascularization Induced byStromal-Derived Factor-1{alpha}. Am J Pathol 1999;154:1125-1135.
95. Mirshahi F, Pourtau J, Li H, et al. SDF-1 activity on microvascular endothelial cells: consequences on angiogenesis in in vitro and in vivo models. Thromb Res 2000;99:587-594.
96. Deshane J, Chen S, Caballero S, et al. Stromal cell-derived factor 1 promotes angiogenesis via a heme oxygenase 1-dependent mechanism. J Exp Med 2007;204:605-618.
97. Aghi M, Cohen KS, Klein RJ, Scadden DT, Chiocca EA. Tumor Stromal-Derived Factor-1 Recruits Vascular Progenitors to Mitotic Neovasculature, where Microenvironment Influences Their Differentiated Phenotypes. Cancer Res 2006;66:9054-9064.
98. Kryczek I, Lange A, Mottram P, et al. CXCL12 and Vascular Endothelial Growth Factor Synergistically Induce Neoangiogenesis in Human Ovarian Cancers. Cancer Res 2005;65:465-472.
99.王雨生.脉络膜新生血管性疾病.人民卫生出版社2007;142.
100. Folkman J, Haudenschild C. Angiogenesis in vitro. Nature 1980;288:551-556.
101. Ruger BM, Breuss J, Hollemann D, et al. Vascular morphogenesis by adult bone marrow progenitor cells in three-dimensional fibrin matrices. Differentiation 2008;76:772-783.
102. Rookmaaker MB, Verhaar MC, Loomans CJM, et al. CD34+ Cells Home, Proliferate, and Participate in Capillary Formation, and in Combination With CD34- Cells Enhance Tube Formation in a 3-Dimensional Matrix. Arterioscler Thromb Vasc Biol 2005;25:1843-1850.
103. Koolwijk P, van Erck MG, de Vree WJ, et al. Cooperative effect of TNFalpha, bFGF, and VEGF on the formation of tubular structures of human microvascular endothelial cells in a fibrin matrix. Role of urokinase activity. J Cell Biol 1996;132:1177-1188.
104.王雨生,严密,杨抚华.视网膜色素上皮细胞培养技术及其应用.中华眼底病杂志1994;10:124-128.
105. Schmetterer L, Weigert G, Luksch A, et al. Reduced Choroidal Blood Flow Parameters Are a Risk Factor for the Development of Neovascular AMD. Invest Ophthalmol Vis Sci 2007;48:5108-.
106. Wang GL, Jiang BH, Semenza GL. Effect of altered redox states on expression and DNA-binding activity of hypoxia-inducible factor 1. Biochem Biophys Res Commun 1995;212:550-556.
107. Zhu J, Wang Y-S, Zhang J, et al. Focal adhesion kinase signaling pathway participates inthe formation of choroidal neovascularization and regulates the proliferation and migration of choroidal microvascular endothelial cells by acting through HIF-1 and VEGF expression in RPE cells. Experimental Eye Research 2008;In Press, Corrected Proof.
108. Arden GB, Wolf JE. Differential Effects of Light and Alcohol on the Electro-oculographic Responses of Patients with Age-Related Macular Disease. Invest Ophthalmol Vis Sci 2003;44:3226-3232.
109. Asahara T, Masuda H, Takahashi T, et al. Bone marrow origin of endothelial progenitor cells responsible for postnatal vasculogenesis in physiological and pathological neovascularization. Circ Res 1999;85:221-228.
110. Kawamoto A, Gwon HC, Iwaguro H, et al. Therapeutic potential of ex vivo expanded endothelial progenitor cells for myocardial ischemia. Circulation 2001;103:634-637.
111. Shintani S, Murohara T, Ikeda H, et al. Mobilization of endothelial progenitor cells in patients with acute myocardial infarction. Circulation 2001;103:2776-2779.
112. Shirakawa K, Shibuya M, Heike Y, et al. Tumor-infiltrating endothelial cells and endothelial precursor cells in inflammatory breast cancer. Int J Cancer 2002;99:344-351.
113. Grant MB, May WS, Caballero S, et al. Adult hematopoietic stem cells provide functional hemangioblast activity during retinal neovascularization. Nat Med 2002;8:607-612.
114. Mohle R, Bautz F, Rafii S, Moore MAS, Brugger W, Kanz L. The Chemokine Receptor CXCR-4 Is Expressed on CD34+ Hematopoietic Progenitors and Leukemic Cells and Mediates Transendothelial Migration Induced by Stromal Cell-Derived Factor-1. Blood 1998;91:4523-4530.
115. Wright DE, Bowman EP, Wagers AJ, Butcher EC, Weissman IL. Hematopoietic Stem Cells Are Uniquely Selective in Their Migratory Response to Chemokines. J Exp Med 2002;195:1145-1154.
116. Shen H, Cheng T, Olszak I, et al. CXCR-4 Desensitization Is Associated with Tissue Localization of Hemopoietic Progenitor Cells. J Immunol 2001;166:5027-5033.
117. Ruger BM, Breuss J, Hollemann D, et al. Vascular morphogenesis by adult bone marrow progenitor cells in three-dimensional fibrin matrices. Differentiation 2007;0.
118. Xiao Q, Kiechl S, Patel S, et al. Endothelial progenitor cells, cardiovascular risk factors, cytokine levels and atherosclerosis--results from a large population-based study. PLoS ONE 2007;2:e975.