人外周血平滑肌祖细胞与成熟血管平滑肌细胞生物学特性比较
|
摘要
目的：探讨成人外周血来源的平滑肌祖细胞(smooth muscle progenitor cells,SPCs)与成熟血管平滑肌细胞(smooth muscle cells, SMCs)细胞形态、增殖潜能等方面的异同点,为血管组织工程提供可靠的种子细胞来源。
方法：1、密度梯度离心法分离成人外周血中单个核细胞,于用纤粘连蛋白包被的含血小板衍生生长因子BB (Platelet derived growth factor BB, PDGF-BB)和EGM-2培养基的培养瓶中诱导培养,观察细胞形态变化,常规换液、传代、冻存及复苏。
2、通过手术取得实验用大隐静脉3cm,采用胶原蛋白酶消化、差异贴壁法原代培养,含PDGF-BB的EGM-2培养基培养,观察细胞形态,获得平滑肌细胞。
3、平滑肌祖细胞及平滑肌细胞分别进行细胞爬片,应用细胞免疫荧光检测平滑肌肌动蛋白(a-SMA)和调宁蛋白(calponinl)的表达。
4、用RT-PCR检测平滑肌肌动蛋白.(alpha-smooth muscle actin, a-sma)和调宁蛋白(calponin 1, cap)的mRNA表达。
5、MTT(四唑盐比色实验)分别检测第6代平滑肌祖细胞与平滑肌细胞的增殖能力。
结果：1、SPCs传代细胞培养达80-90%融合时,呈螺旋状排列,并可呈典型“峰、谷”样形态,符合平滑肌细胞形态学特征。
2、平滑肌祖细胞可表达平滑肌细胞特异性标记α-SMA和calponinl。
3、第6代的SPCs(n=10, A=0.43±0.02)增殖能力明显强于第6代的SMCs(n=10, A=0.21±0.02, P<0.01)。
结论：1、外周血中单个核细胞在PDGF-BB的刺激下可以分化为具有平滑肌细胞特性的平滑肌祖细胞。
2、平滑肌祖细胞较平滑肌细胞增殖旺盛,培养3个月未见老化,具有发展为-种理想的血管组织工程种子细胞来源的可能性。
Objective:Compared the cell morphology and proliferation potential of smooth muscle progenitor cells(SPCs) derived from human peripheral blood with mature vessel wall smooth muscle cells(SMCs) to provide a reliable source of seed cells.
Methods:1. Used Ficoll density gradient centrifugation to separate the mononuclear cells from adult peripheral blood. Cultured in the fibronectin-coated culture flask,and add EGM-2 medium which contails platelet-derived growth factor(PDGF-BB).Observed the morphological changes, liquid passage, cryopreservation and resuscitation by conventional methods.
2. Obtained experimental vena saphena magna 3cm through the surgery, uses the collagen proteinase digestion, the difference to paste the wall law original generation of raise, raise in the EGM-2 culture medium including PDGF-BB. observation the change of cellular form, obtains the smooth muscle cell.
3. Smooth muscle progenitor cells and smooth muscle cells were cell-climb filmed to detect the expression of a-SMA and calponinl by immunofluorescence.
4. Reverse transcription-polymerase chain reaction detected the mRNA expression of smooth muscle actin (a-SMA) and calponinl.
5. MTT detected the sixth generation proliferation of the smooth muscle progenitor cells and smooth muscle cell.
Results:1. SPCs culture passage up to 80-90% confluence, appearing spiral arrangement,and presenting typical "peak, valley" form, sign smooth muscle cell morphology.
2. Smooth muscle progenitor cells can express smooth muscle cell specific marker a-SMA and calponinl.
3. The sixth generation of SPCs (n=10, A=0.43±0.02) proliferation was significantly faster than the same generations of SMCs (n=10, A=0.21±0.02, P <0 .01).
Conclusion:1.Peripheral blood mononuclear cells in the stimulus of platelet-derived growth factor can differentiate into smooth muscle progenitor cells.
2.Compared to smooth muscle cells, smooth muscle progenitor cells proliferate better, and not to see the aging in 3 months cultivation. Smooth muscle progenitor cells has the development of the possibility which one kind of ideal blood vessel organization project seed cell originates.
方法：1、密度梯度离心法分离成人外周血中单个核细胞,于用纤粘连蛋白包被的含血小板衍生生长因子BB (Platelet derived growth factor BB, PDGF-BB)和EGM-2培养基的培养瓶中诱导培养,观察细胞形态变化,常规换液、传代、冻存及复苏。
2、通过手术取得实验用大隐静脉3cm,采用胶原蛋白酶消化、差异贴壁法原代培养,含PDGF-BB的EGM-2培养基培养,观察细胞形态,获得平滑肌细胞。
3、平滑肌祖细胞及平滑肌细胞分别进行细胞爬片,应用细胞免疫荧光检测平滑肌肌动蛋白(a-SMA)和调宁蛋白(calponinl)的表达。
4、用RT-PCR检测平滑肌肌动蛋白.(alpha-smooth muscle actin, a-sma)和调宁蛋白(calponin 1, cap)的mRNA表达。
5、MTT(四唑盐比色实验)分别检测第6代平滑肌祖细胞与平滑肌细胞的增殖能力。
结果：1、SPCs传代细胞培养达80-90%融合时,呈螺旋状排列,并可呈典型“峰、谷”样形态,符合平滑肌细胞形态学特征。
2、平滑肌祖细胞可表达平滑肌细胞特异性标记α-SMA和calponinl。
3、第6代的SPCs(n=10, A=0.43±0.02)增殖能力明显强于第6代的SMCs(n=10, A=0.21±0.02, P<0.01)。
结论：1、外周血中单个核细胞在PDGF-BB的刺激下可以分化为具有平滑肌细胞特性的平滑肌祖细胞。
2、平滑肌祖细胞较平滑肌细胞增殖旺盛,培养3个月未见老化,具有发展为-种理想的血管组织工程种子细胞来源的可能性。
Objective:Compared the cell morphology and proliferation potential of smooth muscle progenitor cells(SPCs) derived from human peripheral blood with mature vessel wall smooth muscle cells(SMCs) to provide a reliable source of seed cells.
Methods:1. Used Ficoll density gradient centrifugation to separate the mononuclear cells from adult peripheral blood. Cultured in the fibronectin-coated culture flask,and add EGM-2 medium which contails platelet-derived growth factor(PDGF-BB).Observed the morphological changes, liquid passage, cryopreservation and resuscitation by conventional methods.
2. Obtained experimental vena saphena magna 3cm through the surgery, uses the collagen proteinase digestion, the difference to paste the wall law original generation of raise, raise in the EGM-2 culture medium including PDGF-BB. observation the change of cellular form, obtains the smooth muscle cell.
3. Smooth muscle progenitor cells and smooth muscle cells were cell-climb filmed to detect the expression of a-SMA and calponinl by immunofluorescence.
4. Reverse transcription-polymerase chain reaction detected the mRNA expression of smooth muscle actin (a-SMA) and calponinl.
5. MTT detected the sixth generation proliferation of the smooth muscle progenitor cells and smooth muscle cell.
Results:1. SPCs culture passage up to 80-90% confluence, appearing spiral arrangement,and presenting typical "peak, valley" form, sign smooth muscle cell morphology.
2. Smooth muscle progenitor cells can express smooth muscle cell specific marker a-SMA and calponinl.
3. The sixth generation of SPCs (n=10, A=0.43±0.02) proliferation was significantly faster than the same generations of SMCs (n=10, A=0.21±0.02, P <0 .01).
Conclusion:1.Peripheral blood mononuclear cells in the stimulus of platelet-derived growth factor can differentiate into smooth muscle progenitor cells.
2.Compared to smooth muscle cells, smooth muscle progenitor cells proliferate better, and not to see the aging in 3 months cultivation. Smooth muscle progenitor cells has the development of the possibility which one kind of ideal blood vessel organization project seed cell originates.
引文
[1]黄铿,齐威力.组织工程化软骨种子细胞的研究与展望[J].中国临床康复,2005；9(10)：172-174.
[2]吴莹琛,崔磊,曹谊林.组织工程血管研究进展[J].组织工程与重建外科杂志,2006；2(5)：288-290.
[3]Yeh ET, Zhang S, Wu HD, et al. Transdifferentiation of human peripheral blood CD34+ enriched cell population into cardiomyocytes, endothelial cells, and smooth muscle cells in vivo[J]. Circulation,2003; 108(17):2070-3.
[4]Simper D, Stalboerger PG, Panetta CJ, et al. Smooth muscle progenitor cells in human blood[J]. Circulation,2002;106(10):1199-204.
[5]Kashlwakura Y,Katoh Y,Tamayose K,etal.Isolation of bone marrow stromal cell-derived smooth muscle cells by a human SM22a promoter:in vitro diferentiation of putative smooth muscle progenitor cells of bone marrow[J]. Circulation,2003,107:2078-2081.
[6]Beltrami S,Golub JS,AmitM,etal.Endothelial cells derived from human emb ryonic stern cells[J].PNAS,2002,99:4391-4396.
[7]Caplice NM, Bunch TJ, Stalboerger PG, et al. Smooth muscle cells in human coronary atherosclerosis can originate from cells administered at marrow transplantation[J]. Proc Natl Acad Sci USA,2003;100(8):4754-9.
[8]Sata M. Role of circulating vascular progenitors in angiogenesis, vascular healing, and pulmonary hypertension:lessons from animal models[J]. Arterioscler Thromb Vasc Biol,2006;26(5):1008-14.
[9]Liu C, Nath KA, Katusic ZS, et al. Smooth muscle progenitor cells in vascular disease [J].Trends Cardiovasc Med,2004;14(7):288-93.
[10]Zoll J, Fontaine V, Gourdy P, et al. Role of human smooth muscle cell progenitors in atherosclerotic plaque development and composition[J]. Cardiovasc Res,2008;77(3): 471-80.
[11]Satoh K, Berk BC. Circulating smooth muscle progenitor cells:novel players in plaquestability[J]. Cardiovasc Res,2008;77(3):445-7.
[12]Lee OK, Coathup MJ, Goodship AE, et al.Use of mesenchymal stem cells to facilitate bone regeneration in normal andchemotherapy treated rats[J].Tissue Eng 2005; 11(11-12): 1727-1735
[13]Defoul A, Perkins GL, Hal J DJ, et al.Glucocorticoids promote chondrogenic diferentiation of adult human mesenchymal stem cells by enhancing expression of cartilage extracellular matrix genes [J].Stem Cells 2006;24(6):1487-1495.
[14]吴少峰,黄惠民.组织工程血管研究的新进展[J].生物医学与临床,2004；8(1)：49-52.
[15]唐新华,黄玲,夏宁等.血管组织工程种子细胞原代培养技术的改良[J].中国组织工程研究与临床康复,009；12(42),8277-8280.
[16]Heylingers JM, Arts CH,Verhangen HJ,etal.Improving small diameter vascular grafts:from applicationg of an endothelial cell lining to the construction of a tissue engineered blood vessel [J]. Ann Vasc Surg.2005; 19:448-456.
[17]Lu SJ,Ivanovs Y,Feng Q,etal. Hemangioblasts from human embryonic stem cells generate multilayered blood vessels with functional smooth muscle cells[J].Reqen Med,2009; 4(1):37-47.
[18]Yamashita J, Itoh H, HirashimaM, et al. Flkl-positive cells derived from embryonic stem cells serve as vascular progenitors [J].Nature,2000;408(6808):92-96.
[19]Kyba M,Perlingeiro RC,Hoover RR,etal.Enhanced fematopoietic differentiation of embryonic stem cells conditionally expressing Stat5[J].Proc Natl Sci USA,2003; 100(7); 11904-11910.
[20]郭泽君,边云飞,薛君,等.体外培养成人脂肪间充质干细胞及向平滑肌细胞诱导分化[J].中国动脉硬化杂志,2009；17(3)：189-192.
[21]Gajrowthdusek CM,Luo Z,Mayberg MR.Basic fibroblast growth fator and transforming growth factor β1:Synergistic mediators of angiogenesia in vitro[J].Cell Physiol,1993; 157(12):133-149.
[22]Cairrao E, Santos-Silva AJ, Alvarez E,etal.Isolation and culture of human umbilical artery smooth muscle cells expressing functional calcium channels[J]. In Vitro Cell Dev Biol Anim.2009,45(3-4):175-84.
[23]Hoch R V,Soriano P. Roles of PDGF in animal development[J]. Development,2003; 130:4679284.
[24]Dandre F,Owens G K. Platelet-derived growth factor-BB and Ets-1 transcription factor negatively regulate transcription of multiple smooth muscle cell differentiation marker genes [J].Am J Physiol Heart Circ Physiol,2004;286:20422-2051.
[25]McKinnon R D,Waldron S, Kiel M E. PDGF alpha-receptor signal strength controls an RTK rheostat that integrates phosphoinositol 3'-kinase and paospholipase Cgamma pathways during oligodendrocyte maturation [J]. J Neurosci,2005;25(14):3499-3508.
[26]王云雅,汪静,邓敬兰,等.放射性PDGFR反义寡核苷酸对血管平滑肌细胞增殖和凋亡的影响[J].心脏杂志,2006；18(4)：388-390.
[27]Werth C, Stuhlmann D, Cat B,et al. Stromal resistance of fibroblasts against oxidative damage:involvement of tumorcell-secreted platelet derived growth factor (PDGF) and phosphoinositide 32kinase (PI3K) activation [J]. Carcinogenesis,2008;3 [Epub ahead of print].
[28]Francy J M,Nag A,Concroy E J,et al. Sphingosine kinase 1 expression is regulated by signaling through PI3K, AKT2 and mTOR in human coronary artery smooth muscle cells [J]. Biochim Biophys Acta,2007;1769(4):253-265.
[29]李英,段惠军.Raf/MEK/ERK信号转导通路在糖尿病肾病发生发展中的作用[J].国际 内分泌代谢杂志,2007；27(7)：269-2711.
[30]Nakata S, Fujita N,Kitagawa Y,et al. Regulation of platelet2derived growth factor receptor activation by afadin through SHP-2:implications for cellular morphology [J]. J Biol Chem,2007;282 (52):37815-37825.
[31]Sandilands E,Cans C, Fincham V J,et al. RhoB and actin polymerization coordinate Src activation with endosome mediated delivery to the membrane[J]. Dev Cell,2004; 7(6):855-869.
[32]Kojima N, Hori M, Murata T, et al. Different profiles of Ca2+responses to endothelin-1 and PDGF in liver myofibroblasts during the process of cell differentiation[J].Br J Pharmacol,2007;151 (6):816-827.
[33]Egan G G,Wainwright C L,Wadsworth R W,et al. PDGF induced signaling in proliferating and differentiated vascular smooth muscle:effects of altered intracellular Ca(2+) regulation[J]. Cardiovasc Res,2005;67(2):308-316.
[34]Ducasse E, Cosset J M, Eschwege F, et al. Hyperplasia of the arterial intima due to smooth muscle cell proliferation. Current data,experimental treatments and perspectives [J]. M al Vasc,2003;28(3):130-44.
[35]Fukuda D, Sata M, Tanaka K, et al. Potent inhibitory effect of sirolimus on circulating vascular progenitor cells [J]·Circulation,2005;111 (7):926-931.
[36]Caplice NM, Doyle B. Vascular progenitor cells:origin and mechanisms of mobilization, differentiation integration, and vasculogenesis[J]. Stem Cells Dev.2005;14(2):122-139.
[37]Hayashi S,W atanabe N,Nakazawa K,etal.Roles of Pselectin in inflammation. neointimal formation and vascular remodeling in balloon-injured rat carotid arteries[J]. Circulation, 2000; 102(14):1710-1717.
[38]Grimm PC, Nickerson P, Jeffery J, etal. Neointimal and tubulolnterstitial infiltration by recipient mesenchymal cells in chronic renal-allograft rejection[J]. N Engl J Med,2001; 345(2):93-97.
[39]Saiura A.Sata M,Hirata Y,et al.Circulating smooth muscle progenitor cells contribute to atherosclerosis[J].Nat Med,2001;7(4):382-383.
[40]Schober A, Hoffmann R, Opree N, et al. Peripheral CD34+cells and the risk of in-stent restenosis in patients with coronary heart disease. Am J Cardiol.2005;96(8):1116-1122.
[41]Tanaka K, Sata M, Hirata Y, et al. Diverse contribution of bonemarrow cells to neointimal hyperplasis after mechanical vascular Injuries[J]. Circ Res.2003;93(8):783-790.
[42]Law RE, Meehan WP, Xi XP, et al. Troglitazone inhibits vascular smooth muscle cell growth and intimal hyperp lasia[J]. J Clin Invest,1996;98 (8):1897-1905.
[1]David Simper, Paul G.,Stalboerger,etal.Smooth Muscle Progenitor Cells in Human Blood[J]. Circulation 2002; 106:1199-1204;
[2]Sata M, Saiura A, Kunisato A, Tojo A, etal. Hematopoietic stem cells differentiateinto vascular cells that participate in thepathogenesis of atherosclerosis[J].Nat Med.2002;8:403-409.
[3]Ducasse E, Cosset J M, Eschwege F, etal. Hyperplasia of the arterial intima due to smooth muscle cell proliferation. Current data,experimental treatments and perspectives [J]. M al Vase,2003,28(3):130-44.
[4]Nie L,Wise M L,Peterson D M, etal. Avenanthramide, a polyphenol from oats, inhibits vascular smooth muscle cell proliferation and enhances nitric oxide production[J] Atherosclerosis,2006,186(2):260-266.
[5]Sugiyama S, Kugiyama K, Nakamura S, et al. Characterization of smooth muscle-like cells in circulating human peripheral blood[J]. Atherosclerosis.2006,187(2):351-62.
[6]Keith A. Hruska.Vascular Smooth Muscle Cells in the Pathogenesis of Vascular Calcification[J]. Circulation Research.2009; 104:710-711
[7]Sata M. Role of circulating vascular progenitors in angiogenesis, vascular healing, and pulmonary hypertension:lessons from animal models[J]. Arterioscler Thromb Vase Biol, 2006,26(5):1008-14.
[8]Satoh K, Berk BC. Circulating smooth muscle progenitor cells:novel players in plaque stability[J]. Cardiovasc Res JT-Cardiovascular research,2008,77(3):445-7.
[9]SataM, SaiuraA, NagaiR, eta.l. Hematopoietic stem cells differentiate into vascular cells that participate in the pathogenesis of atherosclerosis[J]. NatMed,2002,8:403-409.
[10]SimperD,StalboergerPG,CapliceNM,etal.Smoothmuscle progenitor cells in human blood[J]. Circulation,2002,106:1199-1204.
[11]Kashlwakura Y,Katoh Y,Tamayose K,etal.Isolation of bone marrow stromal cell-derived smooth muscle cells by a human SM22a promoter:in vitro diferentiation of putative smooth muscle progenitor cells of bone marrow[J]. Circulation,2003,107:2078-2081.
[12]Bentzon JF, Weile C, Falk E, etal. Smoothmuscle cells in atherosclerosis originate from the local vessel wall and not circulating progenitor cells in ApoE knockoutmice[J]. ArteriosclerThromb Vasc Biol,2006,26:2696-2702.
[13]Hu Y,Davison F,Xu Q,etal.Smooth muscle cells in transplant atherosclerotic lesions are originated from recipients, but not bone marrow progenitor cells[J]. Circulation,2002,106: 1834-1839.
[14]Beltrami S,Golub JS,AmitM,etal.Endothelial cells derived from human emb ryonic stern cells[J].PNAS,2002,99:4391-4396.
[15]SataM, SaiuraA, NagaiR, etal. Hematopoietic stem cells differentiate into vascular cells that participate in the pathogenesis of atherosclerosis[J]. NatMed,2002,8:403-409.
[16]Yeh ET, Zhang S, Wu HD, et al. Trans differentiation of human peripheral blood CD34+ enriched cell population into cardiomyocytes,endothelial cells,and smooth muscle cel in vivo [J]. Circulation,2003,108(17),2070-207.
[17]Hayashi S,W atanabe N,Nakazawa K,etal.Roles of Pselectin in inflammation. neointimal formation and vascular remodeling in balloon-injured rat carotid arteries[J]. Circulation, 2000,102(14):1710-1717.
[18]Grimm PC, Nickerson P, Jeffery J, etal. Neointimal and tubulolnterstitial infiltration by recipient mesenchymal cells in chronic renal-allograft rejection[J]. N Engl J Med,2001, 345(2):93-97.
[19]Saiura A,Sata M,Hirata Y,et al.Circulating smooth muscle progenitor cells contribute to atherosclerosis[J].Nat Med,2001,7(4):382-383.
[20]Tanaka K,Sata M,Hirata Y,et al.Diverse contribution of bone marrow cells to neointlmal hyperplasia after mechanical vascular Injuries[J]. Circ Res,2003,93(8):783-790.
[21]Sasaguri Y, Wang KY,Tanimoto A,etal.Role of histamine produced by bone marrow derived vascular cells in pathogenesis of atherosclerosis[J].Circ Res,2005,96(9):974-981.
[22]Miller AM, McPhaden AR,Wadsworth RM,etal.Inhibition by leukocyte depletion of neointima formation after balloon angioplasty in a rabbit model of restenosis[J].Cardio vase Res,2001,49(4):838-850.
[23]Furukawa Y, Matsumori A, Ohashi N,etal.Anti-mono-cyte chemoattractant protein-1/ monocyte chemotactic and activating factor antibody inhibits neointimal hyperplasia in injured rat carotid arteries[J].Circ Res,1999,84(3):306-314.
[24]Caplice N M, Bunch T J, Stalboerger PG, et al. Smooth muscle cells in human coronary atherosclerosis can originate from cells administered at marrow transplantation[J]. Proc Natl Acad Sci USA,2003;100(8):4754-9.
[2]吴莹琛,崔磊,曹谊林.组织工程血管研究进展[J].组织工程与重建外科杂志,2006；2(5)：288-290.
[3]Yeh ET, Zhang S, Wu HD, et al. Transdifferentiation of human peripheral blood CD34+ enriched cell population into cardiomyocytes, endothelial cells, and smooth muscle cells in vivo[J]. Circulation,2003; 108(17):2070-3.
[4]Simper D, Stalboerger PG, Panetta CJ, et al. Smooth muscle progenitor cells in human blood[J]. Circulation,2002;106(10):1199-204.
[5]Kashlwakura Y,Katoh Y,Tamayose K,etal.Isolation of bone marrow stromal cell-derived smooth muscle cells by a human SM22a promoter:in vitro diferentiation of putative smooth muscle progenitor cells of bone marrow[J]. Circulation,2003,107:2078-2081.
[6]Beltrami S,Golub JS,AmitM,etal.Endothelial cells derived from human emb ryonic stern cells[J].PNAS,2002,99:4391-4396.
[7]Caplice NM, Bunch TJ, Stalboerger PG, et al. Smooth muscle cells in human coronary atherosclerosis can originate from cells administered at marrow transplantation[J]. Proc Natl Acad Sci USA,2003;100(8):4754-9.
[8]Sata M. Role of circulating vascular progenitors in angiogenesis, vascular healing, and pulmonary hypertension:lessons from animal models[J]. Arterioscler Thromb Vasc Biol,2006;26(5):1008-14.
[9]Liu C, Nath KA, Katusic ZS, et al. Smooth muscle progenitor cells in vascular disease [J].Trends Cardiovasc Med,2004;14(7):288-93.
[10]Zoll J, Fontaine V, Gourdy P, et al. Role of human smooth muscle cell progenitors in atherosclerotic plaque development and composition[J]. Cardiovasc Res,2008;77(3): 471-80.
[11]Satoh K, Berk BC. Circulating smooth muscle progenitor cells:novel players in plaquestability[J]. Cardiovasc Res,2008;77(3):445-7.
[12]Lee OK, Coathup MJ, Goodship AE, et al.Use of mesenchymal stem cells to facilitate bone regeneration in normal andchemotherapy treated rats[J].Tissue Eng 2005; 11(11-12): 1727-1735
[13]Defoul A, Perkins GL, Hal J DJ, et al.Glucocorticoids promote chondrogenic diferentiation of adult human mesenchymal stem cells by enhancing expression of cartilage extracellular matrix genes [J].Stem Cells 2006;24(6):1487-1495.
[14]吴少峰,黄惠民.组织工程血管研究的新进展[J].生物医学与临床,2004；8(1)：49-52.
[15]唐新华,黄玲,夏宁等.血管组织工程种子细胞原代培养技术的改良[J].中国组织工程研究与临床康复,009；12(42),8277-8280.
[16]Heylingers JM, Arts CH,Verhangen HJ,etal.Improving small diameter vascular grafts:from applicationg of an endothelial cell lining to the construction of a tissue engineered blood vessel [J]. Ann Vasc Surg.2005; 19:448-456.
[17]Lu SJ,Ivanovs Y,Feng Q,etal. Hemangioblasts from human embryonic stem cells generate multilayered blood vessels with functional smooth muscle cells[J].Reqen Med,2009; 4(1):37-47.
[18]Yamashita J, Itoh H, HirashimaM, et al. Flkl-positive cells derived from embryonic stem cells serve as vascular progenitors [J].Nature,2000;408(6808):92-96.
[19]Kyba M,Perlingeiro RC,Hoover RR,etal.Enhanced fematopoietic differentiation of embryonic stem cells conditionally expressing Stat5[J].Proc Natl Sci USA,2003; 100(7); 11904-11910.
[20]郭泽君,边云飞,薛君,等.体外培养成人脂肪间充质干细胞及向平滑肌细胞诱导分化[J].中国动脉硬化杂志,2009；17(3)：189-192.
[21]Gajrowthdusek CM,Luo Z,Mayberg MR.Basic fibroblast growth fator and transforming growth factor β1:Synergistic mediators of angiogenesia in vitro[J].Cell Physiol,1993; 157(12):133-149.
[22]Cairrao E, Santos-Silva AJ, Alvarez E,etal.Isolation and culture of human umbilical artery smooth muscle cells expressing functional calcium channels[J]. In Vitro Cell Dev Biol Anim.2009,45(3-4):175-84.
[23]Hoch R V,Soriano P. Roles of PDGF in animal development[J]. Development,2003; 130:4679284.
[24]Dandre F,Owens G K. Platelet-derived growth factor-BB and Ets-1 transcription factor negatively regulate transcription of multiple smooth muscle cell differentiation marker genes [J].Am J Physiol Heart Circ Physiol,2004;286:20422-2051.
[25]McKinnon R D,Waldron S, Kiel M E. PDGF alpha-receptor signal strength controls an RTK rheostat that integrates phosphoinositol 3'-kinase and paospholipase Cgamma pathways during oligodendrocyte maturation [J]. J Neurosci,2005;25(14):3499-3508.
[26]王云雅,汪静,邓敬兰,等.放射性PDGFR反义寡核苷酸对血管平滑肌细胞增殖和凋亡的影响[J].心脏杂志,2006；18(4)：388-390.
[27]Werth C, Stuhlmann D, Cat B,et al. Stromal resistance of fibroblasts against oxidative damage:involvement of tumorcell-secreted platelet derived growth factor (PDGF) and phosphoinositide 32kinase (PI3K) activation [J]. Carcinogenesis,2008;3 [Epub ahead of print].
[28]Francy J M,Nag A,Concroy E J,et al. Sphingosine kinase 1 expression is regulated by signaling through PI3K, AKT2 and mTOR in human coronary artery smooth muscle cells [J]. Biochim Biophys Acta,2007;1769(4):253-265.
[29]李英,段惠军.Raf/MEK/ERK信号转导通路在糖尿病肾病发生发展中的作用[J].国际 内分泌代谢杂志,2007；27(7)：269-2711.
[30]Nakata S, Fujita N,Kitagawa Y,et al. Regulation of platelet2derived growth factor receptor activation by afadin through SHP-2:implications for cellular morphology [J]. J Biol Chem,2007;282 (52):37815-37825.
[31]Sandilands E,Cans C, Fincham V J,et al. RhoB and actin polymerization coordinate Src activation with endosome mediated delivery to the membrane[J]. Dev Cell,2004; 7(6):855-869.
[32]Kojima N, Hori M, Murata T, et al. Different profiles of Ca2+responses to endothelin-1 and PDGF in liver myofibroblasts during the process of cell differentiation[J].Br J Pharmacol,2007;151 (6):816-827.
[33]Egan G G,Wainwright C L,Wadsworth R W,et al. PDGF induced signaling in proliferating and differentiated vascular smooth muscle:effects of altered intracellular Ca(2+) regulation[J]. Cardiovasc Res,2005;67(2):308-316.
[34]Ducasse E, Cosset J M, Eschwege F, et al. Hyperplasia of the arterial intima due to smooth muscle cell proliferation. Current data,experimental treatments and perspectives [J]. M al Vasc,2003;28(3):130-44.
[35]Fukuda D, Sata M, Tanaka K, et al. Potent inhibitory effect of sirolimus on circulating vascular progenitor cells [J]·Circulation,2005;111 (7):926-931.
[36]Caplice NM, Doyle B. Vascular progenitor cells:origin and mechanisms of mobilization, differentiation integration, and vasculogenesis[J]. Stem Cells Dev.2005;14(2):122-139.
[37]Hayashi S,W atanabe N,Nakazawa K,etal.Roles of Pselectin in inflammation. neointimal formation and vascular remodeling in balloon-injured rat carotid arteries[J]. Circulation, 2000; 102(14):1710-1717.
[38]Grimm PC, Nickerson P, Jeffery J, etal. Neointimal and tubulolnterstitial infiltration by recipient mesenchymal cells in chronic renal-allograft rejection[J]. N Engl J Med,2001; 345(2):93-97.
[39]Saiura A.Sata M,Hirata Y,et al.Circulating smooth muscle progenitor cells contribute to atherosclerosis[J].Nat Med,2001;7(4):382-383.
[40]Schober A, Hoffmann R, Opree N, et al. Peripheral CD34+cells and the risk of in-stent restenosis in patients with coronary heart disease. Am J Cardiol.2005;96(8):1116-1122.
[41]Tanaka K, Sata M, Hirata Y, et al. Diverse contribution of bonemarrow cells to neointimal hyperplasis after mechanical vascular Injuries[J]. Circ Res.2003;93(8):783-790.
[42]Law RE, Meehan WP, Xi XP, et al. Troglitazone inhibits vascular smooth muscle cell growth and intimal hyperp lasia[J]. J Clin Invest,1996;98 (8):1897-1905.
[1]David Simper, Paul G.,Stalboerger,etal.Smooth Muscle Progenitor Cells in Human Blood[J]. Circulation 2002; 106:1199-1204;
[2]Sata M, Saiura A, Kunisato A, Tojo A, etal. Hematopoietic stem cells differentiateinto vascular cells that participate in thepathogenesis of atherosclerosis[J].Nat Med.2002;8:403-409.
[3]Ducasse E, Cosset J M, Eschwege F, etal. Hyperplasia of the arterial intima due to smooth muscle cell proliferation. Current data,experimental treatments and perspectives [J]. M al Vase,2003,28(3):130-44.
[4]Nie L,Wise M L,Peterson D M, etal. Avenanthramide, a polyphenol from oats, inhibits vascular smooth muscle cell proliferation and enhances nitric oxide production[J] Atherosclerosis,2006,186(2):260-266.
[5]Sugiyama S, Kugiyama K, Nakamura S, et al. Characterization of smooth muscle-like cells in circulating human peripheral blood[J]. Atherosclerosis.2006,187(2):351-62.
[6]Keith A. Hruska.Vascular Smooth Muscle Cells in the Pathogenesis of Vascular Calcification[J]. Circulation Research.2009; 104:710-711
[7]Sata M. Role of circulating vascular progenitors in angiogenesis, vascular healing, and pulmonary hypertension:lessons from animal models[J]. Arterioscler Thromb Vase Biol, 2006,26(5):1008-14.
[8]Satoh K, Berk BC. Circulating smooth muscle progenitor cells:novel players in plaque stability[J]. Cardiovasc Res JT-Cardiovascular research,2008,77(3):445-7.
[9]SataM, SaiuraA, NagaiR, eta.l. Hematopoietic stem cells differentiate into vascular cells that participate in the pathogenesis of atherosclerosis[J]. NatMed,2002,8:403-409.
[10]SimperD,StalboergerPG,CapliceNM,etal.Smoothmuscle progenitor cells in human blood[J]. Circulation,2002,106:1199-1204.
[11]Kashlwakura Y,Katoh Y,Tamayose K,etal.Isolation of bone marrow stromal cell-derived smooth muscle cells by a human SM22a promoter:in vitro diferentiation of putative smooth muscle progenitor cells of bone marrow[J]. Circulation,2003,107:2078-2081.
[12]Bentzon JF, Weile C, Falk E, etal. Smoothmuscle cells in atherosclerosis originate from the local vessel wall and not circulating progenitor cells in ApoE knockoutmice[J]. ArteriosclerThromb Vasc Biol,2006,26:2696-2702.
[13]Hu Y,Davison F,Xu Q,etal.Smooth muscle cells in transplant atherosclerotic lesions are originated from recipients, but not bone marrow progenitor cells[J]. Circulation,2002,106: 1834-1839.
[14]Beltrami S,Golub JS,AmitM,etal.Endothelial cells derived from human emb ryonic stern cells[J].PNAS,2002,99:4391-4396.
[15]SataM, SaiuraA, NagaiR, etal. Hematopoietic stem cells differentiate into vascular cells that participate in the pathogenesis of atherosclerosis[J]. NatMed,2002,8:403-409.
[16]Yeh ET, Zhang S, Wu HD, et al. Trans differentiation of human peripheral blood CD34+ enriched cell population into cardiomyocytes,endothelial cells,and smooth muscle cel in vivo [J]. Circulation,2003,108(17),2070-207.
[17]Hayashi S,W atanabe N,Nakazawa K,etal.Roles of Pselectin in inflammation. neointimal formation and vascular remodeling in balloon-injured rat carotid arteries[J]. Circulation, 2000,102(14):1710-1717.
[18]Grimm PC, Nickerson P, Jeffery J, etal. Neointimal and tubulolnterstitial infiltration by recipient mesenchymal cells in chronic renal-allograft rejection[J]. N Engl J Med,2001, 345(2):93-97.
[19]Saiura A,Sata M,Hirata Y,et al.Circulating smooth muscle progenitor cells contribute to atherosclerosis[J].Nat Med,2001,7(4):382-383.
[20]Tanaka K,Sata M,Hirata Y,et al.Diverse contribution of bone marrow cells to neointlmal hyperplasia after mechanical vascular Injuries[J]. Circ Res,2003,93(8):783-790.
[21]Sasaguri Y, Wang KY,Tanimoto A,etal.Role of histamine produced by bone marrow derived vascular cells in pathogenesis of atherosclerosis[J].Circ Res,2005,96(9):974-981.
[22]Miller AM, McPhaden AR,Wadsworth RM,etal.Inhibition by leukocyte depletion of neointima formation after balloon angioplasty in a rabbit model of restenosis[J].Cardio vase Res,2001,49(4):838-850.
[23]Furukawa Y, Matsumori A, Ohashi N,etal.Anti-mono-cyte chemoattractant protein-1/ monocyte chemotactic and activating factor antibody inhibits neointimal hyperplasia in injured rat carotid arteries[J].Circ Res,1999,84(3):306-314.
[24]Caplice N M, Bunch T J, Stalboerger PG, et al. Smooth muscle cells in human coronary atherosclerosis can originate from cells administered at marrow transplantation[J]. Proc Natl Acad Sci USA,2003;100(8):4754-9.