ATRA诱导下MSCs在损伤肺组织的招募及对MMP-9 mRNA的影响
摘要
目的:对全反式维甲酸(All-trans retinoic acid,ATRA)诱导骨髓间充质干细胞(Mesenchymal stem cells,MSCs)在损伤肺组织的招募情况同组织中基质金属蛋白酶9信使核糖核酸(Matrix metalloproteinase 9 mRNA,MMP-9 mRNA)的表达情况进行相关性的研究。
方法:对实验动物随机给予MSCs及ATRA的方式,分别于2周,3周,4周后取肺组织切片,激光共聚焦显微镜下观察植入细胞的招募情况,RT-PCR法检测组织中MMP-9 mRNA的表达变化。
结果:随着时间的延长,在各时间点:在实验组,可见较多经过标记后的MSCs分布于肺泡表面,与MSCs移植组,ATRA干预组及病例对照组相比较其积分光密度(Integral optical density,IOD)明显增强,差异有统计学意义(P<0.05)。同时MMP-9 mRNA在病例对照组及ATRA干预组较高水平表达,而MSCs移植组中随着MSCs移植后时间延长,其表达水平随之降低,比较差异有统计学意义(P<0.05),而实验组MMP-9 mRNA在2周,3周,4周表达又较MSCs移植组低,两者比较差异有统计学意义(P<0.05)。
结论:ATRA对肺损伤组织中移植MSCs的招募作用的影响与组织中MMP-9 mRNA水平的降低有关。
Objective:To research the correlation between the effect of all-trans retinoic acid(ATRA) on recruiting MSCs and MMP-9 mRNA expression in damage tissue.
Method:MSCs and ATRA were separately used in experimental SD rats.At 2 weeks,3 weeks and 4 weeks after the treatment,RT-PCR assay was used to determine expressions of MMP-9 mRNA in lung tissues.Laser scanning microscope was used to observe the recruitment of implanted MSCs.
Result:We found that,at each time point,experimental group had more implanted MSCs in the surface of alveoli and increased IOD than other groups.There was significant difference in these groups(P<0.05).At the same time,in control group and ATRA intervention group,there were overexpression of MMP-9 mRNA,but in MSCs implanted group the expression of MMP-9 mRNA decreased slowly,as compared with the above two groups(P<0.05).In experimental group,the drop of MMP-9 mRNA expression was clear,as compared with the above three groups(P<0.05).
Conclusion:The effect of ATRA on recruitiment of MSCs had relationship with the drop of MMP-9 mRNA expression in damage tissue.
方法:对实验动物随机给予MSCs及ATRA的方式,分别于2周,3周,4周后取肺组织切片,激光共聚焦显微镜下观察植入细胞的招募情况,RT-PCR法检测组织中MMP-9 mRNA的表达变化。
结果:随着时间的延长,在各时间点:在实验组,可见较多经过标记后的MSCs分布于肺泡表面,与MSCs移植组,ATRA干预组及病例对照组相比较其积分光密度(Integral optical density,IOD)明显增强,差异有统计学意义(P<0.05)。同时MMP-9 mRNA在病例对照组及ATRA干预组较高水平表达,而MSCs移植组中随着MSCs移植后时间延长,其表达水平随之降低,比较差异有统计学意义(P<0.05),而实验组MMP-9 mRNA在2周,3周,4周表达又较MSCs移植组低,两者比较差异有统计学意义(P<0.05)。
结论:ATRA对肺损伤组织中移植MSCs的招募作用的影响与组织中MMP-9 mRNA水平的降低有关。
Objective:To research the correlation between the effect of all-trans retinoic acid(ATRA) on recruiting MSCs and MMP-9 mRNA expression in damage tissue.
Method:MSCs and ATRA were separately used in experimental SD rats.At 2 weeks,3 weeks and 4 weeks after the treatment,RT-PCR assay was used to determine expressions of MMP-9 mRNA in lung tissues.Laser scanning microscope was used to observe the recruitment of implanted MSCs.
Result:We found that,at each time point,experimental group had more implanted MSCs in the surface of alveoli and increased IOD than other groups.There was significant difference in these groups(P<0.05).At the same time,in control group and ATRA intervention group,there were overexpression of MMP-9 mRNA,but in MSCs implanted group the expression of MMP-9 mRNA decreased slowly,as compared with the above two groups(P<0.05).In experimental group,the drop of MMP-9 mRNA expression was clear,as compared with the above three groups(P<0.05).
Conclusion:The effect of ATRA on recruitiment of MSCs had relationship with the drop of MMP-9 mRNA expression in damage tissue.
引文
[1]Massova I,Kotra LP,Fridman R,e1 a1.Matrix metalloproteinases:Structure,evolution, and diversification [J].FASEB J,1998:12(12):1075-1095
[2]Woessner J F.Matrix metalloproteinase and their inhibitors in connective tissue remodeling[J].FASEB J,200l:5(8):2l45-2154
[3] Arthur Bank.Hematopoietic stem cell gene therapy:selecting only the best[J].J Clin Invest,2003:112(10):1478-1480
[4]Doherty MM,Liu J,randell SH,et al.Phenotype and differentiation potential of a novel rat tracheal epithelial cell line[J].Am J Respir Cell Mol Biol,1995: 12(4): 385-395
[5] A Delplanque,C Coraux,R Tirouvanziam,et al.Epithelial stem cell mediated development of the human respiratory mucosa in SCID mice[J].Journal Cell Science, 2000:113(5):767-778
[6] RJ Mason,Mc Williams,HL Moses,et al.Stem cells in lung devdopment,disease,and therapy[J].Am J Respir Cell Mol Biol,1997:16(4):355-363
[7]Otto WR.Lung epithelial stem cells [J].J Pathol,2002:197(4):527-535
[8]Juan R,Sanchez-Ramos.Neural cells derived from adult bone marrow and umbilical cord blood [J].Journal of Neuroscicnce Research,2002:69(6):880- 893
[9]Jiang Y,Vaessen B,Lenvik T,et al.Multipotent progenitor cells can be isolated from postnatal murine bone narrow, muscle, and brain [J].Exp Hematol,2002:30(8):896-904
[10]Ratajczak MZ,Kucia M,Reca R,et al.Stem cell plasticity revisited: CXCR4-positive cells expressing mRNA for early muscle, liver and neural cells‘hide out’in the bone marrow[J].Leukemia,2004:18:29-40
[11]Ceradini DJ,Kulkarni AR,Callaghan MJ,et al.Progenitor cell trafficking is regulated by hypoxic gradients through HIF-1 induction of SDF-1[J].Nature Medicine, 2004: 10:858-864
[12]Yano T,Deterding R,Simonet WS,et al.Keratinocyte growth factor reduces lungdamage due to acid instillation in rats[J].Am J Respir Cell Mol Biol,1996:15(4):433-442
[13]Ulrike Bartram,Christian P Speer,The role of transforming growth factorβi?n lung development and disease[J].Chest,2004:125(2):754-765
[14]Tremain N,Korkko J,Ibberson D,et al.MicroSAGE analysis OF 2.353 expressed genes in a single cell-derived colony of undifferentiated human mesenchymal stem cells reveals mRNAs of multiple cell lineages[J].Stem Cells,2001:19(5):408-418
[15]C.C.Yen,S.H.Yang,C.Y.Lin,et al.Stem cells in the lung parenchyma and prospects for lung injury therapy[J].European Journal of Clinical Investigation,2006:36(5):310-319
[16]Massaro GD,Chanbon P.Retinoic acid receptor alpha regulates pulmonary alveolus formation in mice after but not during the perinatal period[J].Am J Physiol, 2003: 284(2):431-433
[17] Wouter G,Mailkel P,Maria W,et al.Neuronal differentiation of embryonic stem cells [J].Molecular Cell Research,1996:12(5):21-26
[18]Tepper J,Juergen p,Melinda A,et al.Can retinoic acid ameliorate the physidogic and morphologyic effects of elastase of elastase instillation in the rat [J]. Chest, 2000: 117(5):242-244
[19]Gross P,Pfitzer E A,Tolker E,et al.Exprimental amphysema:its production with papain in normal and slicotic rats[J].Arch Environ Health,1965 Jul,11(1):50-58
[20]李红梅,崔德健,佟欣,等.熏香烟加气管注内毒素和单纯熏香烟法建立大鼠COPD模型[J].中国病理生理杂志.2002,18(7):808-812
[21] Barnes PJ.Mechanisms in C0PD:differences from asthma[J].Chest Feb,2000: 117(2):1O-14
[22]Claire Legrand,Christine Gilles,Jean-Marie Zahm,et al.Airway epithelial cell migration dynamics:MMP-9 role in cell-extracellular matrix remodeling[J].Cell Biol, 1999:146(2):517-522
[23]Krause DS,Theise ND,Collector MI,et al.Multi-organ,multi-lineage engraftemnt by a single bone marrow-derived stem cell[J].Cell 2001:105:369-377
[24]Ishizawa K,Kubo H,Yamada M,et al.Bone marrow-derived cells contribute to lung regeneration after elastase-induced pulmonary emphysema[J].FEBS Letters, 2004V: 556:249-252
[25]Tepper J,Juergen P,Melinda A,et al.Can retinoic acid ameliorate the physidogic and morphologyic effects of elastase instillation in the rat[J].Chest May,2000:117:242-244
[26]付小兵,孙晓庆,孙同柱,等.表皮细胞生长因子治疗创面出现的干细胞岛现象[J].中华医学杂志.2001:81(12):733-736
[27]Cuenca-López MD, Zamora-Navas P, García-Herrera JM, et al.Adult stem cells applied to tissue engineering and regenerative medicine[J]. Cell Mol Biol, 2008; 54(1): 40-51
[28]Egi K,Conrad NE,Kwan J,el a1.Inhibition of inducible nitric oxide synthase and superoxide production reduces matrix metalloproteinase-9 activity and restores coronary vasomotor function in rat cardiac allografts[J].Eur J Cardiothorac Surg.2004:26(2):262-269
[29]RenóF,Baj G,Surico N,et a1.Exogenous pmstaglandin E2 inhibits TPA induced matrix metalloproteinase-9 production in MCF-7 cells[J].Prostaglandins other lipid mediat,2004: 73(3-4):237-2473
[30]0kada Y,Watanabe S,Nakanashi I,et al Znactivation of tissue inhibitor of metalloproteinaes by neutrophil elastase and other serine proteinases[J].FEBSlett,1988,229(1):157～60
[1]魏宁,陈昶,高文等.干细胞诱导分化肺组织的研究进展[J] .中华结核和呼吸杂志,2006:29(4):270-272
[2]C.C.Yen,S.H.Yang,C.Y.Lin,et al.Stem cells in the lung parenchyma and prospects for lung injury therapy[J].European Journal of Clinical Investigation,2006:36(5):310-319
[3]Doherty MM,Liu J,randell SH,et al.Phenotype and differentiation potential of a novel rat tracheal epithelial cell line[J].Am J Respir Cell Mol Biol,1995:12(4):385-395
[4] A Delplanque,C Coraux,R Tirouvanziam,et al.Epithelial stem cell mediated development of the human respiratory mucosa in SCID mice[J].Journal Cell Science, 2000:113(5):767-778
[5] RJ Mason,Mc Williams,HL Moses,et al.Stem cells in lung devdopment,disease,and therapy[J].Am J Respir Cell Mol Biol,1997:16(4):355-363
[6]Otto WR.Lung epithelial stem cells [J].J Pathol,2002:197(4):527-535
[7] SI Danto,JM Shannon,Z Borok,et al.Reversible transdifferentiation of alveolar epithelial cells[J]. Am J Respir Cell Mol Biol,1995:12(5):497-502
[8] J.M.Shannon,S.D.Jennings,L.D.Nielsen.Modulation of alveolar type II cell differentiated function in vitro[J].Am J Physiol Lung Cell Mol Physiol,1992:262: 427-436
[9]郭春宝,孙波.干细胞与肺[J].国外医学儿科学分册,2003:30(6):283-285
[10] Arthur Bank.Hematopoietic stem cell gene therapy:selecting only the best[J].J Clin Invest,2003:112(10):1478-1480
[11]Juan R,Sanchez-Ramos.Neural cells derived from adult bone marrow and umbilical cord blood [J].Journal of Neuroscicnce Research,2002:69(6):880- 893
[12]Jiang Y,Vaessen B,Lenvik T,et al.Multipotent progenitor cells can be isolated from postnatal murine bone narrow, muscle, and brain [J].Exp Hematol,2002:30(8):896-904
[13]Tremain N,Korkko J,Ibberson D,et al.MicroSAGE analysis OF 2.353 expressed genes in a single cell-derived colony of undifferentiated human mesenchymal stem cells reveals mRNAs of multiple cell lineages[J].Stem Cells,2001:19(5):408-418
[14] Hess DC,Hill WD,Carroll JE,et al.Do bone marrow cells generate neurons ?[J]. Archives of Neurology,2004:61(4):483-485.
[15]Jori FP,Napolitano MA,Melone MA,et al.Role of RB and RB2/P130 genes in marrow stromal stem cells plasticity [J].J Cell Physiol,2004:200(2): 201-212
[16]林群,雷立华,曾邦雄等.骨髓间充质干细胞介导大鼠肺内基因转移的可行性[J] .中华麻醉学杂志,2006: 26(3) : 230-233
[17]Neuringer P,Randel S.Stem cell and repair of lung injuries[J].Respir Res,2004:5:6
[18]Ratajczak MZ,Kucia M,Reca R,et al.Stem cell plasticity revisited: CXCR4-positive cells expressing mRNA for early muscle, liver and neural cells‘hide out’in the bone marrow[J].Leukemia,2004:18:29-40
[19]Ceradini DJ,Kulkarni AR,Callaghan MJ,et al.Progenitor cell trafficking is regulated by hypoxic gradients through HIF-1 induction of SDF-1[J].Nature Medicine, 2004: 10:858-864
[20]Yano T,Deterding R,Simonet WS,et al.Keratinocyte growth factor reduces lung damage due to acid instillation in rats[J].Am J Respir Cell Mol Biol,1996:15(4):433-442
[21]Ulrike Bartram,Christian P Speer,The role of transforming growth factorβi?n lung development and disease[J].Chest,2004:125(2):754-765
[22] Wouter G,Mailkel P,,Maria W,et al.Neuronal differentiation of embryonic stem cells [J].Molecular Cell Research,1996:12(5):21-26
[23]Massaro GD,Chanbon P.Retinoic acid receptor alpha regulates pulmonary alveolus formation in mice after but not during the perinatal period[J].Am J Physiol, 2003: 284(2):431-433
[24]Tepper J,Juergen p,Melinda A,et al.Can retinoic acid ameliorate the physidogic and morphologyic effects of elastase of elastase instillation in the rat[J].Chest, 2000: 117(5):242-244
[25]Daniel Rubio,Javier Garcia-Castro,Mar′?a C,et al.Spontaneous Human Adult Stem Cell Transformation[J].Cancer Research,2005:65(8):3035-3039
[26]Maria Ester Bernardo,Nadia Zaffaroni,Francesca Novara,et al.Human Bone Marrow-Derived Mesenchymal Stem Cells Do Not Undergo Transformation after Long-term In vitro Culture and Do Not Exhibit Telomere Maintenance Mechanisms[J]. Cancer Research,2007:67:9142-9149
[27]Bernardo ME,Avanzini MA,Perotti C,et al.Optimization of in vitro expansion of human multipotent mesenchymal stromal cells for cell-therapy approaches:further insights in the search for a fetal calf serum substitute[J].J Cell Physiol,2007: 211(1): 121-130
[28]Engelhardt JF,Yankaskas JR,Ernst SA,et al.Submucosal glands are the predominant site of CFTR expression in the human bronchus[J].Nature Genetics,1992:2:240-248
[2]Woessner J F.Matrix metalloproteinase and their inhibitors in connective tissue remodeling[J].FASEB J,200l:5(8):2l45-2154
[3] Arthur Bank.Hematopoietic stem cell gene therapy:selecting only the best[J].J Clin Invest,2003:112(10):1478-1480
[4]Doherty MM,Liu J,randell SH,et al.Phenotype and differentiation potential of a novel rat tracheal epithelial cell line[J].Am J Respir Cell Mol Biol,1995: 12(4): 385-395
[5] A Delplanque,C Coraux,R Tirouvanziam,et al.Epithelial stem cell mediated development of the human respiratory mucosa in SCID mice[J].Journal Cell Science, 2000:113(5):767-778
[6] RJ Mason,Mc Williams,HL Moses,et al.Stem cells in lung devdopment,disease,and therapy[J].Am J Respir Cell Mol Biol,1997:16(4):355-363
[7]Otto WR.Lung epithelial stem cells [J].J Pathol,2002:197(4):527-535
[8]Juan R,Sanchez-Ramos.Neural cells derived from adult bone marrow and umbilical cord blood [J].Journal of Neuroscicnce Research,2002:69(6):880- 893
[9]Jiang Y,Vaessen B,Lenvik T,et al.Multipotent progenitor cells can be isolated from postnatal murine bone narrow, muscle, and brain [J].Exp Hematol,2002:30(8):896-904
[10]Ratajczak MZ,Kucia M,Reca R,et al.Stem cell plasticity revisited: CXCR4-positive cells expressing mRNA for early muscle, liver and neural cells‘hide out’in the bone marrow[J].Leukemia,2004:18:29-40
[11]Ceradini DJ,Kulkarni AR,Callaghan MJ,et al.Progenitor cell trafficking is regulated by hypoxic gradients through HIF-1 induction of SDF-1[J].Nature Medicine, 2004: 10:858-864
[12]Yano T,Deterding R,Simonet WS,et al.Keratinocyte growth factor reduces lungdamage due to acid instillation in rats[J].Am J Respir Cell Mol Biol,1996:15(4):433-442
[13]Ulrike Bartram,Christian P Speer,The role of transforming growth factorβi?n lung development and disease[J].Chest,2004:125(2):754-765
[14]Tremain N,Korkko J,Ibberson D,et al.MicroSAGE analysis OF 2.353 expressed genes in a single cell-derived colony of undifferentiated human mesenchymal stem cells reveals mRNAs of multiple cell lineages[J].Stem Cells,2001:19(5):408-418
[15]C.C.Yen,S.H.Yang,C.Y.Lin,et al.Stem cells in the lung parenchyma and prospects for lung injury therapy[J].European Journal of Clinical Investigation,2006:36(5):310-319
[16]Massaro GD,Chanbon P.Retinoic acid receptor alpha regulates pulmonary alveolus formation in mice after but not during the perinatal period[J].Am J Physiol, 2003: 284(2):431-433
[17] Wouter G,Mailkel P,Maria W,et al.Neuronal differentiation of embryonic stem cells [J].Molecular Cell Research,1996:12(5):21-26
[18]Tepper J,Juergen p,Melinda A,et al.Can retinoic acid ameliorate the physidogic and morphologyic effects of elastase of elastase instillation in the rat [J]. Chest, 2000: 117(5):242-244
[19]Gross P,Pfitzer E A,Tolker E,et al.Exprimental amphysema:its production with papain in normal and slicotic rats[J].Arch Environ Health,1965 Jul,11(1):50-58
[20]李红梅,崔德健,佟欣,等.熏香烟加气管注内毒素和单纯熏香烟法建立大鼠COPD模型[J].中国病理生理杂志.2002,18(7):808-812
[21] Barnes PJ.Mechanisms in C0PD:differences from asthma[J].Chest Feb,2000: 117(2):1O-14
[22]Claire Legrand,Christine Gilles,Jean-Marie Zahm,et al.Airway epithelial cell migration dynamics:MMP-9 role in cell-extracellular matrix remodeling[J].Cell Biol, 1999:146(2):517-522
[23]Krause DS,Theise ND,Collector MI,et al.Multi-organ,multi-lineage engraftemnt by a single bone marrow-derived stem cell[J].Cell 2001:105:369-377
[24]Ishizawa K,Kubo H,Yamada M,et al.Bone marrow-derived cells contribute to lung regeneration after elastase-induced pulmonary emphysema[J].FEBS Letters, 2004V: 556:249-252
[25]Tepper J,Juergen P,Melinda A,et al.Can retinoic acid ameliorate the physidogic and morphologyic effects of elastase instillation in the rat[J].Chest May,2000:117:242-244
[26]付小兵,孙晓庆,孙同柱,等.表皮细胞生长因子治疗创面出现的干细胞岛现象[J].中华医学杂志.2001:81(12):733-736
[27]Cuenca-López MD, Zamora-Navas P, García-Herrera JM, et al.Adult stem cells applied to tissue engineering and regenerative medicine[J]. Cell Mol Biol, 2008; 54(1): 40-51
[28]Egi K,Conrad NE,Kwan J,el a1.Inhibition of inducible nitric oxide synthase and superoxide production reduces matrix metalloproteinase-9 activity and restores coronary vasomotor function in rat cardiac allografts[J].Eur J Cardiothorac Surg.2004:26(2):262-269
[29]RenóF,Baj G,Surico N,et a1.Exogenous pmstaglandin E2 inhibits TPA induced matrix metalloproteinase-9 production in MCF-7 cells[J].Prostaglandins other lipid mediat,2004: 73(3-4):237-2473
[30]0kada Y,Watanabe S,Nakanashi I,et al Znactivation of tissue inhibitor of metalloproteinaes by neutrophil elastase and other serine proteinases[J].FEBSlett,1988,229(1):157～60
[1]魏宁,陈昶,高文等.干细胞诱导分化肺组织的研究进展[J] .中华结核和呼吸杂志,2006:29(4):270-272
[2]C.C.Yen,S.H.Yang,C.Y.Lin,et al.Stem cells in the lung parenchyma and prospects for lung injury therapy[J].European Journal of Clinical Investigation,2006:36(5):310-319
[3]Doherty MM,Liu J,randell SH,et al.Phenotype and differentiation potential of a novel rat tracheal epithelial cell line[J].Am J Respir Cell Mol Biol,1995:12(4):385-395
[4] A Delplanque,C Coraux,R Tirouvanziam,et al.Epithelial stem cell mediated development of the human respiratory mucosa in SCID mice[J].Journal Cell Science, 2000:113(5):767-778
[5] RJ Mason,Mc Williams,HL Moses,et al.Stem cells in lung devdopment,disease,and therapy[J].Am J Respir Cell Mol Biol,1997:16(4):355-363
[6]Otto WR.Lung epithelial stem cells [J].J Pathol,2002:197(4):527-535
[7] SI Danto,JM Shannon,Z Borok,et al.Reversible transdifferentiation of alveolar epithelial cells[J]. Am J Respir Cell Mol Biol,1995:12(5):497-502
[8] J.M.Shannon,S.D.Jennings,L.D.Nielsen.Modulation of alveolar type II cell differentiated function in vitro[J].Am J Physiol Lung Cell Mol Physiol,1992:262: 427-436
[9]郭春宝,孙波.干细胞与肺[J].国外医学儿科学分册,2003:30(6):283-285
[10] Arthur Bank.Hematopoietic stem cell gene therapy:selecting only the best[J].J Clin Invest,2003:112(10):1478-1480
[11]Juan R,Sanchez-Ramos.Neural cells derived from adult bone marrow and umbilical cord blood [J].Journal of Neuroscicnce Research,2002:69(6):880- 893
[12]Jiang Y,Vaessen B,Lenvik T,et al.Multipotent progenitor cells can be isolated from postnatal murine bone narrow, muscle, and brain [J].Exp Hematol,2002:30(8):896-904
[13]Tremain N,Korkko J,Ibberson D,et al.MicroSAGE analysis OF 2.353 expressed genes in a single cell-derived colony of undifferentiated human mesenchymal stem cells reveals mRNAs of multiple cell lineages[J].Stem Cells,2001:19(5):408-418
[14] Hess DC,Hill WD,Carroll JE,et al.Do bone marrow cells generate neurons ?[J]. Archives of Neurology,2004:61(4):483-485.
[15]Jori FP,Napolitano MA,Melone MA,et al.Role of RB and RB2/P130 genes in marrow stromal stem cells plasticity [J].J Cell Physiol,2004:200(2): 201-212
[16]林群,雷立华,曾邦雄等.骨髓间充质干细胞介导大鼠肺内基因转移的可行性[J] .中华麻醉学杂志,2006: 26(3) : 230-233
[17]Neuringer P,Randel S.Stem cell and repair of lung injuries[J].Respir Res,2004:5:6
[18]Ratajczak MZ,Kucia M,Reca R,et al.Stem cell plasticity revisited: CXCR4-positive cells expressing mRNA for early muscle, liver and neural cells‘hide out’in the bone marrow[J].Leukemia,2004:18:29-40
[19]Ceradini DJ,Kulkarni AR,Callaghan MJ,et al.Progenitor cell trafficking is regulated by hypoxic gradients through HIF-1 induction of SDF-1[J].Nature Medicine, 2004: 10:858-864
[20]Yano T,Deterding R,Simonet WS,et al.Keratinocyte growth factor reduces lung damage due to acid instillation in rats[J].Am J Respir Cell Mol Biol,1996:15(4):433-442
[21]Ulrike Bartram,Christian P Speer,The role of transforming growth factorβi?n lung development and disease[J].Chest,2004:125(2):754-765
[22] Wouter G,Mailkel P,,Maria W,et al.Neuronal differentiation of embryonic stem cells [J].Molecular Cell Research,1996:12(5):21-26
[23]Massaro GD,Chanbon P.Retinoic acid receptor alpha regulates pulmonary alveolus formation in mice after but not during the perinatal period[J].Am J Physiol, 2003: 284(2):431-433
[24]Tepper J,Juergen p,Melinda A,et al.Can retinoic acid ameliorate the physidogic and morphologyic effects of elastase of elastase instillation in the rat[J].Chest, 2000: 117(5):242-244
[25]Daniel Rubio,Javier Garcia-Castro,Mar′?a C,et al.Spontaneous Human Adult Stem Cell Transformation[J].Cancer Research,2005:65(8):3035-3039
[26]Maria Ester Bernardo,Nadia Zaffaroni,Francesca Novara,et al.Human Bone Marrow-Derived Mesenchymal Stem Cells Do Not Undergo Transformation after Long-term In vitro Culture and Do Not Exhibit Telomere Maintenance Mechanisms[J]. Cancer Research,2007:67:9142-9149
[27]Bernardo ME,Avanzini MA,Perotti C,et al.Optimization of in vitro expansion of human multipotent mesenchymal stromal cells for cell-therapy approaches:further insights in the search for a fetal calf serum substitute[J].J Cell Physiol,2007: 211(1): 121-130
[28]Engelhardt JF,Yankaskas JR,Ernst SA,et al.Submucosal glands are the predominant site of CFTR expression in the human bronchus[J].Nature Genetics,1992:2:240-248