当归多糖对小鼠骨髓基质细胞黏附分子表达及其黏附功能的影响
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摘要
骨髓基质细胞(bone marrow stromal cell,BMSC)是造血微环境的主要组成部分,一方面通过分泌体液因子调控造血;另一方面,通过细胞间的直接接触作用于造血干/祖细胞(hematopoietic progenitor/stem cell,HSPC)的各个阶段,BMSC与造血细胞之间的直接接触是通过其表面的粘附分子来实现的,对造血细胞的增殖与分化起直接支持作用,并且参与HSPC的动员和归巢,其结构和功能的完整性对于保护机体造血的稳定性具有十分重要的作用。粘附分子分为整合素家族、选择素家族及免疫球蛋白基因超家族等多种。其中细胞间粘附分子-1 (intercellular adhesion molecule-1,ICAM-1,CD54)和血管细胞粘附分子-1 (vascular cell adhesion molecule-1,VCAM-1,CD106)是免疫球蛋白基因超家族中最常表达的两种粘附分子,在介导BMSC对造血细胞的粘附及二者间的信息传递方面起重要作用。VCAM-1和ICAM-1有多种功能:能协助造血细胞粘附定位于利于发育的微环境;参与造血细胞与基质细胞形成多聚体;参与HSPC的动员。近年来,VCAM-1、ICAM-1等黏附分子已成为研究的热点。1
当归作为中医“补血、活血”要药已有数千年的临床应用历史。研究表明:当归多糖(Angelica polysaccharides,APS)是当归中促进造血的有效成分,对正常或贫血小鼠的髓系造血祖细胞(BFU-E、CFU-E、CFU-GM和CFU-MK)的增殖分化有显著的促进作用;同时APS在体外可促进人红系、粒单系及混合系造血祖细胞的增殖与分化。
前期实验已经证实:APS对小鼠HSPC有一定的动员作用,与重组人粒细胞集落刺激因子(recombinate human granulocyte colony- stimulating factor,rhG-CSF)联合动员的效果好于单用APS或单用rhG-CSF的动员效果,并且APS动员的小鼠HSPC能够重建造血衰竭小鼠长期造血。目前关于BMSC在造血干细胞(hematopoietic stem cell, HSC)动员中的作用已有很多报道。但是APS的动员机制还未见报道,其动员作用是否是通过改变粘附分子如ICAM-1和VCAM-1的表达及其黏附功能来实现有待进一步深入探讨。这一研究结果将为改善骨髓造血微环境,促进HSPC的动员提供新的思路。
本研究以BALB/c小鼠为实验动物模型,分为两个部分进行实验研究:1.细胞密度和血清对小鼠BMSC体外生长的影响。2. APS对小鼠BMSC生长、黏附分子表达及其黏附功能的影响。本研究旨在探讨APS促进造血以及参与HSPC动员的可能机制,其深入研究将为进一步开发和利用当归提供理论与实验依据。
1.细胞密度和血清对小鼠BMSC体外生长的影响
目的:研究不同细胞接种密度、血清浓度及种类对小鼠BMSC体外生长的影响。
方法:设置不同细胞密度、不同种类血清及血清量的培养体系,观察小鼠BMSC的贴壁情况,细胞80%贴壁时间,用台盼蓝排斥试验检测活细胞率,MTT法检测细胞增殖情况,HE染色光镜下观察细胞形态。
结果:
1.原代全血培养的小鼠BMSC适宜接种密度为(1～4)×106/ mL。
2.细胞最适生长的血清浓度为10%～20%。
3.与胎牛血清相比马血清对小鼠BMSC的促增殖作用更好(P<0.05)。
4.不同种类血清培养的小鼠BMSC类型有一定差异(P<0.05)。
结论:不同细胞接种密度及血清浓度和血清种类对小鼠BMSC体外生长有一定影响。
2.当归多糖对小鼠BMSC黏附分子表达和黏附功能的影响目的:研究APS对小鼠BMSC增殖、粘附分子ICAM-1和VCAM-1的表达及其对骨髓单个核细胞黏附功能的影响。
方法:观察APS对小鼠BMSC消化时间的影响;MTT法检测APS对小鼠BMSC增殖及对骨髓单个核细胞黏附功能的影响;流式细胞术检测APS体内外对小鼠BMSC表面黏附分子ICAM-1和VCAM-1表达的影响。
结果:
1. 100,200 mg/L APS使BMSC达80%贴壁后消化时间明显缩短(P<0.05)。
2. 50,100 mg/L APS组BMSC增殖能力较强(P<0.05)。
3.体外培养中加入200,300 mg/L APS能使BMSC表面ICAM-1表达降低;加入50,100,200,300 mg/L APS均可使VCAM-1表达降低(P<0.05)。小鼠体内腹腔注射1,2,4,6 mg/kg APS均可使ICAM-1、VCAM-1表达比对照组降低(P<0.05)。
4. 100,200,300 mg/L APS使小鼠BMSC对骨髓单个核细胞黏附率减弱(P<0.05)。
结论:APS能促进小鼠BMSC增殖,降低其表面黏附分子ICAM-1和VCAM-1的表达,进而使小鼠BMSC达80%贴壁后消化时间明显缩短,对骨髓单个核细胞的黏附率减弱。
综上所述,APS能促进小鼠BMSC增殖,降低黏附分子ICAM-1和VCAM-1的表达进而改变BMSC的黏附能力,进一步研究其在改善造血微环境及HSC动员中的作用将为开发和利用当归并为寻找新的动员剂提供实验依据。
Bone marrow stromal cell (BMSC) is an important component of bone marrow microenvironment.It can secrete various cytokines and adhere to hematopoietic cells directly to regulate their proliferation,differentiation mobilization and homing. BMSC’s superficial adhesion molecule involve in direct contactation between BMSC and hematopoietic cell.It is important that their integrality of construction and function protect haematopoietic stability.Adhesion molecule conclude integrin family,selectin family, immunoglobulin gene super family and so on.Intercellular adhesion molecule-1(ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) are the main adhesion molecules of immunoglobulin gene super family.They mediate adhesion between BMSC and hematopoietic cell and assist hematopoietic cell to locate microenvironment.They can also involve in forming multimer of hematopoietic cell and stromal cell and mobilizing hematopoietic stem progenitor cells (HSPC) to peripheral blood. Recently,adhesion molecules of ICAM-1 and VCAM-1are hot research spots.
There are thousands of years that Angelica is an important Chinese crude drug to nourish blood and promote blood flow.Angelica polysaccharides(APS) is one of the main chemical composition in it.The past researches discovered that APS could obviously promote the proliferation and differentiation of BFU-E,CFU-E,CFU-GM and CFU-MK in health and anaemic mice. APS also could promote the proliferation and differentiation of BFU-E,CFU-E,CFU-GM and CFU-Mix in the human being.APS could mobilize HSPC from bone marrow into peripheral blood. And APS in combination with rhG-CSF may cooperate with each other. HSPC mobilized by APS can reconstruct long-term hematopoiesis in hematopoiesis failure mice. Now there are many reports about the effect of BMSC in mobilization of HSPC,but now there is no report about the effect of APS on the expression of ICAM-1 and VCAM-1 in murine BMSC and adhesive rate of murine bone marrow mononuclear cell to BMSC in vitro. This study provide a new idea for improving bone marrow microenvironment and promoting mobilization of HSPC.
In this research experimental animal model is BALB/c mice.There are two parts in this research:
1. Effect of different cell culture density and serums on growth of mice BMSC in vitro.
2. Effect of APS on cell proliferation,the expression of ICAM-1 and VCAM-1 in murine BMSC and adhesive rate of murine bone marrow mononuclear cell to BMSC in vitro.
Our research mainly study that APS improve hematopoietic microenvironment and participate in mobilizable mechanism of HSPC.This findings will provide theoretical and experimental evidence for exploiting and utilizing Angelica.
1. Study on the effect of different cell culture density and serums on growth of mice BMSC in vitro
Objective: To study the effect of different cell culture density and serums on growth of mice BMSC in vitro.
Methods: BMSC were cultured in DMEM/F12 containing different cell density and serums. The rate of adhered cells, cell viability by trypan blue test,proliferation of BMSC by MTT and cell morphological structure under light microscope were observed.
Results: The cultured BMSC proliferated well in DMEM/F12 containing 10%~20% serums and the density of (1~4)×106/mL.It proliferated better in horse serum than fetal bovine serums (P<0.05).Its types were also different in bovine serum,fetal bovine serums and horse serum (P<0.05).
Conclusion: There is influence on the growth of mice BMSC cultured in different cell density and serums.
2. Study on effects of APS on adhesion molecules expression and adhesive function in murine BMSC
Objective: To explore the effect of APS on BMSC’s proliferation,the expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule -1 (VCAM-1) in murine BMSC and adhesive rate of murine bone marrow mononuclear cell to BMSC.
Methods: Observe the effect of APS in digestion time of the murine BMSC under inverted microscope,cell proliferation and the binding capacity of murine bone marrow mononuclear cell to BMSC by MTT assay and the expressions of ICAM-1 and VCAM-l of murine BMSC by flow cytometry.
Results: 1) With 100, 200 mg/L APS, the digestion time of murine BMSC has obviously become shorter compared with control group. 2) The cultured cells proliferated well in DMEM/F12 containing 50 mg/L and 100 mg/L APS. 3) APS of 200 mg/L and 300 mg/L decreased the expression of ICAM-l, APS of 50,100,200 and 300 mg/L APS decreased the expression of VCAM-1.The expression of ICAM-l and VCAM-1 also decreased by intraperitoneal injection of APS.4) With 100,200 and 300 mg/L of APS, the adhesive rate was inferior than control group.
Conclusion: APS can promote BMSC’s proliferation, but it decreases the digestion time of murine BMSC,the expression of ICAM-l and VCAM-1 and the adhesive rate of murine bone marrow mononuclear cell to BMSC.
To sum up, APS can promote BMSC’s proliferation and decreases the expression of ICAM-l and VCAM-1. It will provide experimental evidence for exploiting and utilizing Angelica that APS is further researched in improving bone marrow microenvironment and promoting mobilization of HSPC.
当归作为中医“补血、活血”要药已有数千年的临床应用历史。研究表明:当归多糖(Angelica polysaccharides,APS)是当归中促进造血的有效成分,对正常或贫血小鼠的髓系造血祖细胞(BFU-E、CFU-E、CFU-GM和CFU-MK)的增殖分化有显著的促进作用;同时APS在体外可促进人红系、粒单系及混合系造血祖细胞的增殖与分化。
前期实验已经证实:APS对小鼠HSPC有一定的动员作用,与重组人粒细胞集落刺激因子(recombinate human granulocyte colony- stimulating factor,rhG-CSF)联合动员的效果好于单用APS或单用rhG-CSF的动员效果,并且APS动员的小鼠HSPC能够重建造血衰竭小鼠长期造血。目前关于BMSC在造血干细胞(hematopoietic stem cell, HSC)动员中的作用已有很多报道。但是APS的动员机制还未见报道,其动员作用是否是通过改变粘附分子如ICAM-1和VCAM-1的表达及其黏附功能来实现有待进一步深入探讨。这一研究结果将为改善骨髓造血微环境,促进HSPC的动员提供新的思路。
本研究以BALB/c小鼠为实验动物模型,分为两个部分进行实验研究:1.细胞密度和血清对小鼠BMSC体外生长的影响。2. APS对小鼠BMSC生长、黏附分子表达及其黏附功能的影响。本研究旨在探讨APS促进造血以及参与HSPC动员的可能机制,其深入研究将为进一步开发和利用当归提供理论与实验依据。
1.细胞密度和血清对小鼠BMSC体外生长的影响
目的:研究不同细胞接种密度、血清浓度及种类对小鼠BMSC体外生长的影响。
方法:设置不同细胞密度、不同种类血清及血清量的培养体系,观察小鼠BMSC的贴壁情况,细胞80%贴壁时间,用台盼蓝排斥试验检测活细胞率,MTT法检测细胞增殖情况,HE染色光镜下观察细胞形态。
结果:
1.原代全血培养的小鼠BMSC适宜接种密度为(1～4)×106/ mL。
2.细胞最适生长的血清浓度为10%～20%。
3.与胎牛血清相比马血清对小鼠BMSC的促增殖作用更好(P<0.05)。
4.不同种类血清培养的小鼠BMSC类型有一定差异(P<0.05)。
结论:不同细胞接种密度及血清浓度和血清种类对小鼠BMSC体外生长有一定影响。
2.当归多糖对小鼠BMSC黏附分子表达和黏附功能的影响目的:研究APS对小鼠BMSC增殖、粘附分子ICAM-1和VCAM-1的表达及其对骨髓单个核细胞黏附功能的影响。
方法:观察APS对小鼠BMSC消化时间的影响;MTT法检测APS对小鼠BMSC增殖及对骨髓单个核细胞黏附功能的影响;流式细胞术检测APS体内外对小鼠BMSC表面黏附分子ICAM-1和VCAM-1表达的影响。
结果:
1. 100,200 mg/L APS使BMSC达80%贴壁后消化时间明显缩短(P<0.05)。
2. 50,100 mg/L APS组BMSC增殖能力较强(P<0.05)。
3.体外培养中加入200,300 mg/L APS能使BMSC表面ICAM-1表达降低;加入50,100,200,300 mg/L APS均可使VCAM-1表达降低(P<0.05)。小鼠体内腹腔注射1,2,4,6 mg/kg APS均可使ICAM-1、VCAM-1表达比对照组降低(P<0.05)。
4. 100,200,300 mg/L APS使小鼠BMSC对骨髓单个核细胞黏附率减弱(P<0.05)。
结论:APS能促进小鼠BMSC增殖,降低其表面黏附分子ICAM-1和VCAM-1的表达,进而使小鼠BMSC达80%贴壁后消化时间明显缩短,对骨髓单个核细胞的黏附率减弱。
综上所述,APS能促进小鼠BMSC增殖,降低黏附分子ICAM-1和VCAM-1的表达进而改变BMSC的黏附能力,进一步研究其在改善造血微环境及HSC动员中的作用将为开发和利用当归并为寻找新的动员剂提供实验依据。
Bone marrow stromal cell (BMSC) is an important component of bone marrow microenvironment.It can secrete various cytokines and adhere to hematopoietic cells directly to regulate their proliferation,differentiation mobilization and homing. BMSC’s superficial adhesion molecule involve in direct contactation between BMSC and hematopoietic cell.It is important that their integrality of construction and function protect haematopoietic stability.Adhesion molecule conclude integrin family,selectin family, immunoglobulin gene super family and so on.Intercellular adhesion molecule-1(ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) are the main adhesion molecules of immunoglobulin gene super family.They mediate adhesion between BMSC and hematopoietic cell and assist hematopoietic cell to locate microenvironment.They can also involve in forming multimer of hematopoietic cell and stromal cell and mobilizing hematopoietic stem progenitor cells (HSPC) to peripheral blood. Recently,adhesion molecules of ICAM-1 and VCAM-1are hot research spots.
There are thousands of years that Angelica is an important Chinese crude drug to nourish blood and promote blood flow.Angelica polysaccharides(APS) is one of the main chemical composition in it.The past researches discovered that APS could obviously promote the proliferation and differentiation of BFU-E,CFU-E,CFU-GM and CFU-MK in health and anaemic mice. APS also could promote the proliferation and differentiation of BFU-E,CFU-E,CFU-GM and CFU-Mix in the human being.APS could mobilize HSPC from bone marrow into peripheral blood. And APS in combination with rhG-CSF may cooperate with each other. HSPC mobilized by APS can reconstruct long-term hematopoiesis in hematopoiesis failure mice. Now there are many reports about the effect of BMSC in mobilization of HSPC,but now there is no report about the effect of APS on the expression of ICAM-1 and VCAM-1 in murine BMSC and adhesive rate of murine bone marrow mononuclear cell to BMSC in vitro. This study provide a new idea for improving bone marrow microenvironment and promoting mobilization of HSPC.
In this research experimental animal model is BALB/c mice.There are two parts in this research:
1. Effect of different cell culture density and serums on growth of mice BMSC in vitro.
2. Effect of APS on cell proliferation,the expression of ICAM-1 and VCAM-1 in murine BMSC and adhesive rate of murine bone marrow mononuclear cell to BMSC in vitro.
Our research mainly study that APS improve hematopoietic microenvironment and participate in mobilizable mechanism of HSPC.This findings will provide theoretical and experimental evidence for exploiting and utilizing Angelica.
1. Study on the effect of different cell culture density and serums on growth of mice BMSC in vitro
Objective: To study the effect of different cell culture density and serums on growth of mice BMSC in vitro.
Methods: BMSC were cultured in DMEM/F12 containing different cell density and serums. The rate of adhered cells, cell viability by trypan blue test,proliferation of BMSC by MTT and cell morphological structure under light microscope were observed.
Results: The cultured BMSC proliferated well in DMEM/F12 containing 10%~20% serums and the density of (1~4)×106/mL.It proliferated better in horse serum than fetal bovine serums (P<0.05).Its types were also different in bovine serum,fetal bovine serums and horse serum (P<0.05).
Conclusion: There is influence on the growth of mice BMSC cultured in different cell density and serums.
2. Study on effects of APS on adhesion molecules expression and adhesive function in murine BMSC
Objective: To explore the effect of APS on BMSC’s proliferation,the expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule -1 (VCAM-1) in murine BMSC and adhesive rate of murine bone marrow mononuclear cell to BMSC.
Methods: Observe the effect of APS in digestion time of the murine BMSC under inverted microscope,cell proliferation and the binding capacity of murine bone marrow mononuclear cell to BMSC by MTT assay and the expressions of ICAM-1 and VCAM-l of murine BMSC by flow cytometry.
Results: 1) With 100, 200 mg/L APS, the digestion time of murine BMSC has obviously become shorter compared with control group. 2) The cultured cells proliferated well in DMEM/F12 containing 50 mg/L and 100 mg/L APS. 3) APS of 200 mg/L and 300 mg/L decreased the expression of ICAM-l, APS of 50,100,200 and 300 mg/L APS decreased the expression of VCAM-1.The expression of ICAM-l and VCAM-1 also decreased by intraperitoneal injection of APS.4) With 100,200 and 300 mg/L of APS, the adhesive rate was inferior than control group.
Conclusion: APS can promote BMSC’s proliferation, but it decreases the digestion time of murine BMSC,the expression of ICAM-l and VCAM-1 and the adhesive rate of murine bone marrow mononuclear cell to BMSC.
To sum up, APS can promote BMSC’s proliferation and decreases the expression of ICAM-l and VCAM-1. It will provide experimental evidence for exploiting and utilizing Angelica that APS is further researched in improving bone marrow microenvironment and promoting mobilization of HSPC.
引文
[1] 川芎嗪对同基因骨髓移植小鼠中 VCAM-1/VLA-4 表达的影响[J].中国实验血液学杂志,2005,13(1):20-24.
[2] 三羟异黄酮对辐损伤小鼠骨髓基质细胞的保护作用[J].中国实验血液学杂志,2005,27(6):471-473.
[3] Yoder MC,Williams DA.Matrix molecule interactions with hematopoietic stem cells [J]. Exp-Hematol,1995;23(9):961-967.
[4] Kikuta T,Shimazaki C,Ashihara E,et a1.Mobilization of hematopoietic primitive and committed progenitor cells into blood in mice by anti-vascular adhesion molecule-1 antibody alone or in combination with granulocyte colony-stimulating factor[J].Exp Hemato1,2000;28(3):311-317.
[5] Vermeulen M,Le-Pesteur F,Gagnerault MC,et a1.Role of adhesion molecules in the homing and mobilization of murine hematopoietic stem and progenitor cells[J]. Blood, 1998;92(3):894-900.
[6] Papayannopoulou T,Priestley GV,Nakamoto B,et a1.Molecular pathways in bone marrow homing:dominant role of alpha(4)beta(1)over beta(2)-integrins and selectins[J]. Blood, 2001;98(8):2403-2411.
[7] Montesano R,Schaller G,and Orci L.Induction of epithelial tubular morphogenesis in Vitro by fibroblast-derived soluble Factors[J].Cell,1991; 66(4):697-711.
[8] Van-der-Sluijs JP,Van-den-Bos C,Baert MR,et al.Loss of long-term repopulating ability in long-term bone marrow culture[J].Leukemia, 1993;7(5):725-732.
[9] Boudreau N,Sympson CJ,Werb et al.Suppression of ICE and apoptosis in mammary epithelial cells by extracellular matrix[J].Science, 1995;267(5199):891-893.
[10] Bauer EA. Cell culture density as a modulator of collagenase expression in normal human fibroblast cultures[J].Exp Cell Res,1977;107(2):269-276.
[11] Siczkowski M,Ng LL.Culture density and the activity,abundance and phosphoryla -tion of the Na+/H+ exchanger isoform in human fibroblasts[J]. Biochem Biophys Res Commun,1995;209(1):191-197.
[12] CIemmons DR,Elgin RG and Jams PE.Somatomedin-C binding to cultured human fibroblasts is dependent on donor age and culture density[J].JClin Endocrinol Metab,1986;63(4),996-1001.
[1] Levesque JP,Lli F,Simmons PJ,et al.Characterization of hematopoietic progenitor mobilization in protease-deficient mice[J].Blood,2004,104 (1):65-72.
[2] Gigant C,Latger-Cannard V,Bensoussan et a1.Quantitative expression of the adhesion receptors VLA-4, VLA-5, L-selectin, MAC-1, and ICAM-1 on the surface of CD34+ cells[J]. Transfus Clin Biol,2001,8(6):453-459.
[3] Wysoczynski M,Reca R,Ratajczak J.Incorporation of CXCR4 into membrane lipid rafts primes homing-related responses of hematopoietic stem/progenitor cells to an SDF-1 gradient[J]. Blood,2005,105(1):40-48.
[4] Huang YM,Kucia M,Rezzoug F,et al.Flt3-ligand-mobilized peripheral blood, but not Flt3-ligand-expanded bone marrow, facilitating cells promote establishment of chimerism and tolerance[J].Stem Cells,2006,24 (4):936-948.
[5] 胡晶,吴宏.当归多糖对小鼠外周血造血干细胞动员作用的研究[J].中草药,2006,37(12):1835-1838.
[6] 王改琴,杜小丽,吴宏.细胞密度和血清对小鼠骨髓基质细胞体外生长的影响[J].重庆医科大学学报, 2007,32 (11),1178-1182.
[7] 吴宏,姜蓉等.人参多糖和当归多糖诱导人内皮细胞表达造血生长因子的实验研究[J].中国中西医结合杂志,2002,22(9):687-690.
[8] 李静,王亚平.当归多糖对骨髓巨噬细胞的影响及其与造血调控的关系[J].中草药,2005,36(1):69-72.
[9] Levesque JP,Lli F,Simmons PJ,et al.Characterization of hematopoietic progenitor mobilization in protease-deficient mice[J].Blood,2004,104 (1):65-72.
[1] Seshi B,Kumar S,Sellers D.Human bone marrow stromal cell: coexpression of markers specific for multiple mesenchymal cell lineages[J].Blood Cells Mol Dis, 2000,26(3):234-246.
[2] 唐可京,谢茂灿,李幼姬.细胞间粘附分子-1和血管细胞间粘附分子-1的结构与功能[J].细胞与分子免疫学杂志.2002,18(2):193-195.
[3] Jun CD,Shimaoka M,Carman CV,et a1.Dimerization and the effectiveness ofICAM-1 in mediating LFA-1-dependent adhesion[J].Proc Natl Acad Sci USA. 2001,98(12):6830-6835.
[4] Carion A,Domenech J,Herault O,et a1.Decreased stroma adhesion capacity of CD34+ progenitor cells from mobilized peripheral blood is not lineage-or stage-specific and is associated with low beta 1 and beta 2 integrin expression [J].Hematother Stem Cell Res,2002,11(3):491-500.
[5] Levesque JP,Takamatsu Y,Nilsson SK.Vascular cell adhesion molecule-1 (CD106) is cleaved by neutrophil proteases in the bone marrow following hematopoietic progenitor cell mobilization by granulocyte colony- stimulating factor[J].Blood, 2001,98(5): 1289-1297.
[6] Levesque JP,Lli F,Simmons PJ,et al.Characterization of hematopoietic progenitor mobilization in protease-deficient mice[J].Blood,2004,104 (1):65-72.
[7] Kikuta T,Shimazaki C,Ashihara E,et a1. Mobilization of hematopoietic primitive and committed progenitor cells into blood in mice by anti- vascular adhesion molecule-1 antibody alone or in combination with granulocyte colony-stimulating factor[J].ExpHemato1,2000,28(3)311- 317.
[8] Peled A,Kollet O,Ponomaryov T el a1.The chemokine SDF-1 activates the integrins LFA-1,VLA-4,and VLA-5 on immature human CD34(+) cells: role in transendothelial/stromal migration and engraftment of NOD/SCID mice [J].Blood, 2000,95(11):3289-3296.
[9] Gigant C,Latger-Cannard V,Bensoussan et a1.Quantitative expression of the adhesion receptors VLA-4, VLA-5, L-selectin, MAC-1, and ICAM-1 on the surface of CD34+ cells[J].Transfus Clin Biol,2001,8(6):453-459.
[10] Papayannopoulou T,Priestley GV,Nakamoto B,et al.Molecular pathways in bone marrow homing:dominant role of alpha(4)beta(1) over beta(2)-integrins and selectins.Blood,2001,98(8):2403-2411.
[11] Wysoczynski M,Reca R,Ratajczak J.Incorporation of CXCR4 into membrane hpid rafts primes homing-related responses of hematopoietic stem/ progenitor cells to an SDF-1 gradient [J].Blood,2005,105(1):40-48.
[12] Huang YM,Kucia M,Rezzoug F,et al.Flt3-ligand-mobilized peripheral blood, but not Flt3-ligand-expanded bone marrow, facilitating cells promote establishment of chimerism and tolerance[J].Stem Cells,2006,24 (4): 936-948.
[13] Jack Gold, Helen M.Valinski.Adhesion receptor expression by CD34+ cells from peripheral blood or bone marrow grafts: Correlation with time to engraftment[J]. Experimental Hematology, 2006,34(5):680-687.
[14] 孙岚,刘文励,孙汉英 et a1.川芎嗪对放射损伤小鼠骨髓微环境修复的信号传导机理研究[J].中华放射医学与防护杂志,2003,23(2):93-95.
[15] Gaugler MH, Squiban C, Mouthon MA, et al.Irradiation enhances the support of haemopoietic cell transmigration, proliferation and differentiation by endothelial cells[J].Br J Haematol.2001 113(4):940-950.
[16] Pozo M,Nicolas R,Egido J,et al.Simvastatin inhibits the migration and adhesion of monocytic cells and disorganizes the cytoskeleton of activated endothelial cells[J]. European Journal Of Pharmacology, 2006, 548(24):53-63.
[17] Z. Zhou, D.J. MacEwan.Induction of ICAM-1 and VCAM-1 adhesion molecules in endothelial cells by TNF receptor 1 through the NF-kB1 pathway[J].Vascular Pharmacology,Abstracts,2006,45(3):87-87.
[18] Lupher ML Jr, Harris EA, Beals CR, et a1. Cellular activation of leukocyte function-associated antigen-1 and its affinity are regulated at the I domain allosteric site[J].Immunol,2001, 167(3):1431-1439.
[19] Tatiana Ulyanova,Linda M,Scott,et al.VCAM-1 expression in adult hematopoietic and nonhematopoietic cells is controlled by tissue- inductive signals and reflects their developmental origin[J].Blood, 2005,106(1):86-94.
[20] Fuste B,Escolar G,Marin P.G-CSF increases the expression of VCAM-1 on stromal cells promoting the adhesion of CD34(+) hematopoietic cells: Studies under flow conditions[J]. Experimental Hematology, 2004, 32(8): 765-772.
[21] 黄文荣,王立生.重组人粒细胞集落刺激因子动员对 CD4+ T 淋巴细胞迁移和黏附功能的影响[J].中华血液学杂志,2006,27(2):87-90.
[22] Jenks SA,Miller J.Inhibition of IL-4 responses after T cell priming in the contextof LFA-1 costimulation is not reversed by restimulation in the presence of CD28 costimulation[J].Immunol, 2000,164(1):72-78.
[23] 黄文荣,王立生达万明.T细胞LFA-1/ICAM-1协同刺激信号的研究进展[J].中国实验血液学杂志,2004,12:(4)533-537.
[24] Smits HH,de Jong EC,Schuitemaker JH,et al.Intercellular adhesion molecule-1 /LFA-1 ligation favors human Th1 development[J].Immunol,2002,168(4):1710 -1716.
[2] 三羟异黄酮对辐损伤小鼠骨髓基质细胞的保护作用[J].中国实验血液学杂志,2005,27(6):471-473.
[3] Yoder MC,Williams DA.Matrix molecule interactions with hematopoietic stem cells [J]. Exp-Hematol,1995;23(9):961-967.
[4] Kikuta T,Shimazaki C,Ashihara E,et a1.Mobilization of hematopoietic primitive and committed progenitor cells into blood in mice by anti-vascular adhesion molecule-1 antibody alone or in combination with granulocyte colony-stimulating factor[J].Exp Hemato1,2000;28(3):311-317.
[5] Vermeulen M,Le-Pesteur F,Gagnerault MC,et a1.Role of adhesion molecules in the homing and mobilization of murine hematopoietic stem and progenitor cells[J]. Blood, 1998;92(3):894-900.
[6] Papayannopoulou T,Priestley GV,Nakamoto B,et a1.Molecular pathways in bone marrow homing:dominant role of alpha(4)beta(1)over beta(2)-integrins and selectins[J]. Blood, 2001;98(8):2403-2411.
[7] Montesano R,Schaller G,and Orci L.Induction of epithelial tubular morphogenesis in Vitro by fibroblast-derived soluble Factors[J].Cell,1991; 66(4):697-711.
[8] Van-der-Sluijs JP,Van-den-Bos C,Baert MR,et al.Loss of long-term repopulating ability in long-term bone marrow culture[J].Leukemia, 1993;7(5):725-732.
[9] Boudreau N,Sympson CJ,Werb et al.Suppression of ICE and apoptosis in mammary epithelial cells by extracellular matrix[J].Science, 1995;267(5199):891-893.
[10] Bauer EA. Cell culture density as a modulator of collagenase expression in normal human fibroblast cultures[J].Exp Cell Res,1977;107(2):269-276.
[11] Siczkowski M,Ng LL.Culture density and the activity,abundance and phosphoryla -tion of the Na+/H+ exchanger isoform in human fibroblasts[J]. Biochem Biophys Res Commun,1995;209(1):191-197.
[12] CIemmons DR,Elgin RG and Jams PE.Somatomedin-C binding to cultured human fibroblasts is dependent on donor age and culture density[J].JClin Endocrinol Metab,1986;63(4),996-1001.
[1] Levesque JP,Lli F,Simmons PJ,et al.Characterization of hematopoietic progenitor mobilization in protease-deficient mice[J].Blood,2004,104 (1):65-72.
[2] Gigant C,Latger-Cannard V,Bensoussan et a1.Quantitative expression of the adhesion receptors VLA-4, VLA-5, L-selectin, MAC-1, and ICAM-1 on the surface of CD34+ cells[J]. Transfus Clin Biol,2001,8(6):453-459.
[3] Wysoczynski M,Reca R,Ratajczak J.Incorporation of CXCR4 into membrane lipid rafts primes homing-related responses of hematopoietic stem/progenitor cells to an SDF-1 gradient[J]. Blood,2005,105(1):40-48.
[4] Huang YM,Kucia M,Rezzoug F,et al.Flt3-ligand-mobilized peripheral blood, but not Flt3-ligand-expanded bone marrow, facilitating cells promote establishment of chimerism and tolerance[J].Stem Cells,2006,24 (4):936-948.
[5] 胡晶,吴宏.当归多糖对小鼠外周血造血干细胞动员作用的研究[J].中草药,2006,37(12):1835-1838.
[6] 王改琴,杜小丽,吴宏.细胞密度和血清对小鼠骨髓基质细胞体外生长的影响[J].重庆医科大学学报, 2007,32 (11),1178-1182.
[7] 吴宏,姜蓉等.人参多糖和当归多糖诱导人内皮细胞表达造血生长因子的实验研究[J].中国中西医结合杂志,2002,22(9):687-690.
[8] 李静,王亚平.当归多糖对骨髓巨噬细胞的影响及其与造血调控的关系[J].中草药,2005,36(1):69-72.
[9] Levesque JP,Lli F,Simmons PJ,et al.Characterization of hematopoietic progenitor mobilization in protease-deficient mice[J].Blood,2004,104 (1):65-72.
[1] Seshi B,Kumar S,Sellers D.Human bone marrow stromal cell: coexpression of markers specific for multiple mesenchymal cell lineages[J].Blood Cells Mol Dis, 2000,26(3):234-246.
[2] 唐可京,谢茂灿,李幼姬.细胞间粘附分子-1和血管细胞间粘附分子-1的结构与功能[J].细胞与分子免疫学杂志.2002,18(2):193-195.
[3] Jun CD,Shimaoka M,Carman CV,et a1.Dimerization and the effectiveness ofICAM-1 in mediating LFA-1-dependent adhesion[J].Proc Natl Acad Sci USA. 2001,98(12):6830-6835.
[4] Carion A,Domenech J,Herault O,et a1.Decreased stroma adhesion capacity of CD34+ progenitor cells from mobilized peripheral blood is not lineage-or stage-specific and is associated with low beta 1 and beta 2 integrin expression [J].Hematother Stem Cell Res,2002,11(3):491-500.
[5] Levesque JP,Takamatsu Y,Nilsson SK.Vascular cell adhesion molecule-1 (CD106) is cleaved by neutrophil proteases in the bone marrow following hematopoietic progenitor cell mobilization by granulocyte colony- stimulating factor[J].Blood, 2001,98(5): 1289-1297.
[6] Levesque JP,Lli F,Simmons PJ,et al.Characterization of hematopoietic progenitor mobilization in protease-deficient mice[J].Blood,2004,104 (1):65-72.
[7] Kikuta T,Shimazaki C,Ashihara E,et a1. Mobilization of hematopoietic primitive and committed progenitor cells into blood in mice by anti- vascular adhesion molecule-1 antibody alone or in combination with granulocyte colony-stimulating factor[J].ExpHemato1,2000,28(3)311- 317.
[8] Peled A,Kollet O,Ponomaryov T el a1.The chemokine SDF-1 activates the integrins LFA-1,VLA-4,and VLA-5 on immature human CD34(+) cells: role in transendothelial/stromal migration and engraftment of NOD/SCID mice [J].Blood, 2000,95(11):3289-3296.
[9] Gigant C,Latger-Cannard V,Bensoussan et a1.Quantitative expression of the adhesion receptors VLA-4, VLA-5, L-selectin, MAC-1, and ICAM-1 on the surface of CD34+ cells[J].Transfus Clin Biol,2001,8(6):453-459.
[10] Papayannopoulou T,Priestley GV,Nakamoto B,et al.Molecular pathways in bone marrow homing:dominant role of alpha(4)beta(1) over beta(2)-integrins and selectins.Blood,2001,98(8):2403-2411.
[11] Wysoczynski M,Reca R,Ratajczak J.Incorporation of CXCR4 into membrane hpid rafts primes homing-related responses of hematopoietic stem/ progenitor cells to an SDF-1 gradient [J].Blood,2005,105(1):40-48.
[12] Huang YM,Kucia M,Rezzoug F,et al.Flt3-ligand-mobilized peripheral blood, but not Flt3-ligand-expanded bone marrow, facilitating cells promote establishment of chimerism and tolerance[J].Stem Cells,2006,24 (4): 936-948.
[13] Jack Gold, Helen M.Valinski.Adhesion receptor expression by CD34+ cells from peripheral blood or bone marrow grafts: Correlation with time to engraftment[J]. Experimental Hematology, 2006,34(5):680-687.
[14] 孙岚,刘文励,孙汉英 et a1.川芎嗪对放射损伤小鼠骨髓微环境修复的信号传导机理研究[J].中华放射医学与防护杂志,2003,23(2):93-95.
[15] Gaugler MH, Squiban C, Mouthon MA, et al.Irradiation enhances the support of haemopoietic cell transmigration, proliferation and differentiation by endothelial cells[J].Br J Haematol.2001 113(4):940-950.
[16] Pozo M,Nicolas R,Egido J,et al.Simvastatin inhibits the migration and adhesion of monocytic cells and disorganizes the cytoskeleton of activated endothelial cells[J]. European Journal Of Pharmacology, 2006, 548(24):53-63.
[17] Z. Zhou, D.J. MacEwan.Induction of ICAM-1 and VCAM-1 adhesion molecules in endothelial cells by TNF receptor 1 through the NF-kB1 pathway[J].Vascular Pharmacology,Abstracts,2006,45(3):87-87.
[18] Lupher ML Jr, Harris EA, Beals CR, et a1. Cellular activation of leukocyte function-associated antigen-1 and its affinity are regulated at the I domain allosteric site[J].Immunol,2001, 167(3):1431-1439.
[19] Tatiana Ulyanova,Linda M,Scott,et al.VCAM-1 expression in adult hematopoietic and nonhematopoietic cells is controlled by tissue- inductive signals and reflects their developmental origin[J].Blood, 2005,106(1):86-94.
[20] Fuste B,Escolar G,Marin P.G-CSF increases the expression of VCAM-1 on stromal cells promoting the adhesion of CD34(+) hematopoietic cells: Studies under flow conditions[J]. Experimental Hematology, 2004, 32(8): 765-772.
[21] 黄文荣,王立生.重组人粒细胞集落刺激因子动员对 CD4+ T 淋巴细胞迁移和黏附功能的影响[J].中华血液学杂志,2006,27(2):87-90.
[22] Jenks SA,Miller J.Inhibition of IL-4 responses after T cell priming in the contextof LFA-1 costimulation is not reversed by restimulation in the presence of CD28 costimulation[J].Immunol, 2000,164(1):72-78.
[23] 黄文荣,王立生达万明.T细胞LFA-1/ICAM-1协同刺激信号的研究进展[J].中国实验血液学杂志,2004,12:(4)533-537.
[24] Smits HH,de Jong EC,Schuitemaker JH,et al.Intercellular adhesion molecule-1 /LFA-1 ligation favors human Th1 development[J].Immunol,2002,168(4):1710 -1716.