补肾生血解毒方对免疫介导的再生障碍性贫血小鼠淋巴细胞分化及血液学参数的影响

目的探讨补肾生血解毒方对免疫介导的再生障碍性贫血(AA)小鼠淋巴细胞分化及血液学参数的影响。方法以~(60)Co-γ射线照射结合异源免疫细胞尾静脉注射致小鼠骨髓造血抑制模型,阳性对照药物免疫抑制剂环孢素A、中药补肾生血解毒方干预治疗,连续给药28 d。分别于第7、14、21、28天用血细胞计数仪检测外周血中血液学参数的变化;给药结束后用流式细胞仪检测小鼠外周血细胞中T细胞、B细胞、自然杀伤细胞(NK)比率的改变。结果与模型组相比,环孢素A组与补肾生血解毒方组在第21、28天,小鼠外周血中红细胞(RBC)、血红蛋白(HGB)、血小板(PLT)、白细胞(WBC)数量均升高(P0.05),差异有统计学意义。与模型组相比,补肾生血解毒方组大鼠外周血中CD3~+CD4~+细胞比率增多(P0.01),CD3~-CD19~+细胞和CD3~-NK1.1~+细胞比率降低(P0.01),差异有统计学意义。结论补肾生血解毒方能够通过调控AA小鼠淋巴细胞分化修复骨髓造血功能,达到治疗AA的目的。     

参芪扶正注射液对再生障碍性贫血小鼠脾脏T-bet表达的影响

目的观察参芪扶正注射液对再生障碍性贫血(以下简称再障)小鼠的疗效及其对Th1细胞转录因子T-bet的影响,探讨参芪扶正注射液在再障治疗中的免疫调节作用。方法将小鼠随机分成正常组、模型组及用药组,用药组给予参芪扶正注射液腹腔注射治疗,正常组及模型组给予生理盐水对照治疗,治疗第11天观察外周血、骨髓象及T-bet蛋白表达水平。结果与正常组比较,模型组小鼠外周血象计数、骨髓造血细胞明显降低,T-bet蛋白表达明显升高,差异有统计学意义(P<0.05);与模型组比较,用药组小鼠外周血象计数、骨髓造血细胞明显升高,T-bet表达明显降低,差异有统计学意义(P<0.05)。结论参芪扶正注射液能改善贫血症状,降低再障小鼠脾脏组织中转录因子T-bet的表达水平,逆转Th1型反应,调节免疫异常。     

巴桑母酥油丸对放射线-化学复合损伤小鼠造血干细胞移植后重建造血能力的影响简

目的 研究藏药巴桑母酥油丸对放射线-化学复合损伤后机体骨髓造血干细胞功能的影响,探究其“补益”作用机制.方法 将经60Co γ射线照射＋环磷酰胺腹腔注射方法得到的放射线-化学复合损伤小鼠分为巴桑母酥油丸组、生理盐水组、空白组,分别灌胃巴桑母酥油丸、生理盐水及自然恢复14d后,将其骨髓作为供体移植给受致死剂量γ射线照射的同种受体小鼠,再检测受体小鼠外源性脾集落、骨髓造血祖细胞集落产率、骨髓细胞造血干细胞抗原-1（Sca-1）阳性细胞率,并与灌胃.结果 巴桑母酥油丸组的外源性脾集落、骨髓早期红系祖细胞集落（BFU-E）、粒-巨噬系祖细胞集落（CFU-GM）产率均显著高于空白组和生理盐水组（P＜0.05）,而晚期红系祖细胞集落（CFU-E）、巨核系祖细胞集落（CFU-Meg）、骨髓细胞Sca-1＋细胞率并不离于自然恢复的空白组（P＞0.05）.结论 巴桑母酥油丸可以提高放射线-化学复合损伤小鼠骨髓造血干细胞移植后造血重建能力,提示巴桑母酥油丸可经改善骨髓造血干细胞功能发挥其对放射线-化学复合损伤机体“补益”作用.     

人脐带间充质干细胞对脐血CD34~+细胞在NOD/SCID小鼠体内造血重建的影响

目的 探讨人脐带间充质干细胞对脐血CD34~+细胞在NOD/SCID小鼠体内造血重建的影响.方法 将3.5×10~5个脐血CD34~+细胞单独(单移植组)或与5.0×10~6个人脐带间充质干细胞共同(共移植组)输入经~(137)Cs 3.0Gy照射后的NOD/SCID小鼠体内,观察移植后6周内小鼠外周血象的变化情况.于移植后第6周处死小鼠,采用流式细胞术检测小鼠骨髓、脾脏及外周血人源细胞(hCD45~+)含量,并分别检测小鼠骨髓中人源淋巴系(CD3/CD19)、粒系(CD33)、单核系(CD14)、血小板(CD61)、红系(CD235a)等各系血细胞比例,比较间充质干细胞共移植对CD34~+细胞植入率的影响.结果 移植后3周,两组小鼠外周血象开始有不同程度恢复;移植后6周,共移植组外周血白细胞和血小板计数均已达高峰,明显高于单移植组(P<0.05),两组小鼠的红细胞计数差异无统计学意义(P>0.05).移植后6周,共移植组骨髓及外周血中人源细胞hCD45~+CD34~+比例分别为(42.66±2.57)%和(4.74±1.02)%,明显高于单移植组的(25.27±1.67)%和(1.19±0.54)%(P=0.006).移植后6周,共移植组小鼠骨髓内的CD19~+、CD33~+、CD14~+、CD61~+和CD235a~+细胞比例均明显高于单移植组(P<0.05),CD3~+T淋巴细胞比例明显低于单移植组(P=0.003);CD19~+B淋巴细胞得到优势扩增,明显高于其他各系血细胞比例(P<0.05).结论 脐带间充质干细胞与脐血CD34~+细胞共移植可促进造血干细胞的植入,缩短CD34~+细胞移植后造血恢复时间.     

用胚胎干细胞诱导的造血干/祖细胞重建小鼠造血功能研究

目的 胚胎干细胞体外诱导分化出现CD34^ 造血干/祖细胞，然后将其注射入致死量照射小鼠体内，以研究其重建造血功能。方法 胚胎干细胞自由分化形成胚胎体后，加入造血刺激因子混合液以诱导产生CD34^ 造血干／祖细胞，将这些造血干／祖细胞注射入经致死剂量照射的小鼠，观察小鼠存活率的改变，并取存活2个月的小鼠骨髓和脾脏，PCR检测嵌合体形成。结果 体外分化第13日CD34^ 造血干／祖细胞比例可高达17．36％，将这些细胞注射入致死量照射小鼠后，可提高存活率达86．67％，存活小鼠的骨髓和脾脏均检测到供体来源的细胞。结论 胚胎干细胞体外分化获得CD34^ 造血干/祖细胞，且后者具有其相应的正常生物学功能。     

Effects of Panaxadiol Saponins Component as A New Chinese Patent Medicine on Proliferation, Differentiation and Corresponding Gene Expression Profile of Megakaryocytes

Objective:To investigate the effects of panaxadiol saponins component(PDS-C) isolated from total saponins of panax ginseng on proliferation,differentiation and corresponding gene expression profile of megakaryocytes.Methods:Bone marrow culture of colony forming assay of megakaryocytic progenitor cells(CFU-MK) was observed for the promoting proliferation mediated by PDS-C,and differentiation of megakaryocytic blasts caused by PDS-C was analyzed with flow cytometry in CHRF-288 and Meg-01 cells,as well as proliferation,differentiation-related genes expression profile and protein expression levels were detected by human gene expression microarray and western blot.Results:In response to PDS-C 10,20 and 50 mg/L,CFU-MK from 10 human bone marrow samples was increased by 28.9%±2.7%,41.0%±3.2%and 40.5%±2.6%over untreated control,respectively(P0.01,each).Flow cytometry analysis showed that PDS-C treated CHRF-288 cells and Meg-01 cells significantly increased in CD42 b,CD41,TSP and CD36 positive ratio,respectively.PDS-C induced29 genes up-regulated more than two-fold commonly in both cells detected by human expression microarray representing 4000 known genes.The protein expression levels of ZNF91,c-Fos,BTF3 a,GATA-1,RGS2,NDRG2 and RUNX1 were increased with western blot in correspond to microarray results.Conclusion:PDS-C as an effective component for hematopoiesis,play the role to enhance proliferation and differentiation of megakaryocytes,also up-regulated expression of proliferation,differentiation-related genes and proteins in vitro.     

Effects of glucocorticoid and cysteinyl leukotriene 1 receptor antagonist on CD34+ hematopoietic cells in bone marrow of asthmatic mice

Background Corticosteroids remain the most effective therapy available for asthma. They have widespread effects on asthmatic airway inflammation. However, little is known about the effects of corticosteroids on the production of bone marrow inflammatory cells in asthma. This study observed the effects of glucocorticoid and cysteinyl leukotriene 1 receptor antagonist on CD34+ hematopoietic cells, so as to explore the possible effectiveness of a bone marrow-targeted anti-inflammatory strategy. Methods Balb/c mice were sensitized and challenged with ovalbumin (OVA) to establish an asthmatic model. For two consecutive weeks, asthmatic mice were challenged with OVA while being given either prednisone, montelukast, prednisone plus montelukast, or sterile saline solution. The mice were killed 24 hours after the last challenge with OVA, and bronchoalveolar lavage fluid (BALF), peripheral blood, and bone marrow were collected. Eosinophils in peripheral blood and BALF, and nucleated cells in BALF, peripheral blood, and bone marrow were counted. The percentages of CD34+ cells, CD4+ T lymphocytes and CD8+ T lymphocytes among nucleated cells in peripheral blood and bone marrow were counted by flow cytometry. Immunocytochemistry and in situ hybridization were employed to detect expression of CD34 and interleukin (IL)-5Rα mRNA (CD34+ IL-5Rα mRNA+ cells) among bone marrow hematopoietic cells. Results Compared with the sterile saline solution group, the number of eosinophils in BALF and peripheral blood, CD34+ cells in peripheral blood and bone marrow, and CD34+ IL-5Rα mRNA+ cells in bone marrow of mice from the prednisone and prednisone plus montelukast groups were significantly lower （P&lt;0.01）. The number of eosinophils in BALF from the montelukast group was also significantly lower （P&lt;0.05）. Conclusions The results suggest that, in this asthmatic mouse model, prednisone probably inhibits proliferation, differentiation, and migration of CD34+ cells in bone marrow, blocks eosinophilopoiesis in bone marrow, and interferes with eosinophil migration into peripheral blood and subsequent recruitment in the airway. In addition, montelukast may suppress eosinophil infiltration into the lungs of asthmatic mice. However, a significant inhibitory effect of montelukast on the proliferation and migration of CD34+ cells and a cooperating effect with prednisone on bone marrow of asthmatic mice were not observed.     

rhBMP-2对再生障碍性贫血小鼠骨髓造血恢复的促进作用

探究了人重组骨形态发生蛋白-2(rhBMP-2)对再障小鼠的治疗作用。采用5-FU联合白消安建立小鼠再障模型,通过干预rhBMP-2进行治疗。考察了各实验组小鼠白细胞数、骨髓单核细胞数、体重、存活率、脾系数、粒系-巨系细胞集落(CFU-GM)数以及骨髓单核细胞中CD34⁺细胞比例,进行股骨、脾脏HE切片分析。结果表明,相较于再障对照组,rhBMP-2治疗组小鼠的白细胞数、存活率、CFU-GM集落数及骨髓单核细胞数显著提高,并且骨髓单核细胞中的CD34⁺细胞含量也提高,显著改善脾脏功能和缓解骨髓抑制,证实rhBMP-2能够促进再障小鼠造血损伤的修复。     

Low incidence of severe aGVHD and accelerating hemopoietic reconstitution in allo-BMT using lenograstim stimulated BM cells

Objectives To investigate the efficacy of accelerating hemopoietic reconstraction and reducing a graft versus host disease (GVHD) in Allo-BMT receiving lenogras tim stimulated donor marrow and to assess the preliminary biological mechanism .Methods The donors for thirty patients (study group) with leukemia were given lenograsti m 3-4 μg·kg(-1) ·d(-1) for seven days prior to marrow harvest. The results of subsequent engraftment in the recipients was compared w ith fifteen donors without G-CSF (control group). Five donors themselves were studied t o assess the effects of lenograstion on hematopoietic progenitor cells and lym phocyte subsets in BM.Results The stimulated bone marrow contained a higher number of nucleated cells, CFU-GM and CD34(+) cells (P&lt;0.01). The hematopoetic reconstitution was accelerat ed. Until granulocyte counts exceeded 0.5×10(9) /L and plalete counts exceeded 20×10(9) /L, the days were 16.7±3.2 and 18.4±3.0 days as compared with t hose of the control group (22.5±5.1 and 26.3±5.9 days respectively, P &lt; 0.01). The incidence of grade Ⅱ-Ⅳ aGVHD was very low, only one case with g rade Ⅱ aGVHD on the skin in the study group. Four out of fifteen patients (2 6.7%) in the control group had grade Ⅱ-Ⅳ aGVHD (P&lt;0.05) . The number of T lymphocyte subsets in the harvested BM stimulated by G-CSF changed. In comp arison with the control group, CD4(+) decreased and CD8(+) increased significan tly (P&lt;0.01). The changes of progenitor cells and T lymphocyte subsets in BM from pre- to post- G-CSF stimulation indicated that the percentage o f CD4(+) cells reduced (P&lt;0.05), that of CD8(+) cells, and that of CD34(+) i ncreased (P&lt;0.01). The incidence of chronic GVHD and relapse of leukemia were not different significantly between both groups.Conclusions Allogenic bone marrow transplant (Allo-BMT) donors given G-CSF can accelerate engraftment and minimize the incidence of sever e aGVHD. There is a trend in favour of improved transplant-related complicatians.     

Expression of adherent molecule and cyclin by ligustrazine in bone marrow of mice with immune-induced aplastic anemia

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骨髓干细胞输注修复放疗免疫抑制的动物实验

目的 探讨骨髓干细胞 (bone marrowstem cell, BMSC) 对放疗所致免疫抑制小鼠的修复作用, 为干细胞用于临床修复放化疗后免疫抑制的患者提供一定的理论依据.方法 选择雌性BALB/C小鼠, 随机分为实验组、对照组和空白组, 实验组和对照组的每只小鼠经照射剂量为8 Gy的6 MV直线加速器X线照射, 随后实验组小鼠通过尾静脉输注小鼠骨髓干细胞1×107个, 0.2 m L, 对照组小鼠通过尾静脉输入0.2 m L生理盐水, 空白组小鼠不做任何处理.于移植后第15、25、35天用血细胞分析仪检测外周血血常规, 流式细胞仪检测T淋巴细胞亚群 (CD3+、CD4+、CD8+) 、ELISA试剂盒检测血清免疫球蛋白Ig M变化情况, 并观察生存状态和存活率.结果经放疗所致免疫抑制的小鼠经骨髓干细胞输注后35 d存活率为80%, 未经输注的小鼠35 d存活率为10%;放疗后小鼠经骨髓干细胞输注后外周血白细胞数、淋巴细胞比例及CD3+T细胞亚群较未经输注干细胞的小鼠高 (P<0.05) ;在骨髓干细胞移植后第15天, CD4+%、CD8+%和免疫球蛋白Ig M最低, 随后逐渐升高 (P<0.05) ;骨髓干细胞移植后能降低免疫抑制小鼠外周血单核细胞比例及CD4+/CD8+比值 (P<0.05) .结论 骨髓干细胞的输注能够修复免疫抑制小鼠的免疫功能, 这为干细胞用于临床修复放化疗所致的免疫抑制的病人提供了一定的实验基础和理论依据.     

补肾解毒活血法对环磷酰胺所致小鼠骨髓抑制的预防作用研究

目的研究补肾解毒活血法对环磷酰胺所致骨髓抑制的预防作用及机制。方法昆明小鼠60只,随机分成空白组、模型组和中药预防组,中药预防组连续灌胃给药10 d,空白组和模型组给予等体积的生理盐水。造模后,各组随机抽取10只,于造模前1 d及造模后第1、3、5、7、10天检测外周血象,其余10只小鼠在造模后第1天检测骨髓有核细胞计数、骨髓细胞凋亡情况、造血生长因子及培养造血祖细胞集落。结果造模后外周血WBC、PLT,骨髓有核细胞计数,粒—巨噬系集落形成单位（colony forming unit-granulocyte macrophage,CFU-GM）,造血祖细胞集落形成单位及造血生长因子中药预防组均显著高于模型组（P〈0.01或P〈0.05）。中药预防组总凋亡率远低于模型组（P〈0.01）。结论补肾解毒活血法可以减轻环磷酰胺对小鼠的骨髓抑制毒副作用,明显促进骨髓抑制小鼠WBC、PLT、CFU-GM及骨髓有核细胞的增加,抑制环磷酰胺所致的细胞凋亡。     

参芪扶正注射液对化疗后小鼠造血功能影响的实验研究

目的 探讨参芪扶正注射液对化疗后小鼠造血功能的影响.方法 用5-氟尿嘧啶（5-FU）225 mg/kg腹腔注射复制骨髓抑制动物模型,治疗组给予参芪扶正注射液5 ml/kg,对照组给予同剂量生理盐水.治疗1周后,做血细胞计数、各系造血祖细胞集落（CFU-GM、CFU-E、CFU-MK）培养和骨髓病理检查.结果 参芪扶正注射液能升高外周血细胞计数：治疗组小鼠外周血的红细胞、白细胞和血小板计数均高于对照组（P〈0.05）.促进造血祖细胞增殖：治疗组小鼠的CFU-GM、CFU-E、CFU-MK集落数均大于对照组（P〈0.05）.改善骨髓抑制：骨髓病理检查对照组出现大片空白区,造血细胞稀少,而治疗组造血组织结构较完整,造血细胞量丰富.结论 化疗后小鼠在骨髓抑制、造血功能低下时,参芪扶正注射液能通过促进骨髓各系造血祖细胞的增殖,改善骨髓造血组织增生,从而促进血细胞的生成.     

哮喘小鼠CD34+祖细胞和嗜酸性粒细胞的动态变化及其与CCR3/eotaxin表达的关系

目的 观察不同时间点CD34+祖细胞和嗜酸性粒细胞在哮喘小鼠支气管肺泡灌洗液(BALF)、外周血、骨髓悬液中的动态变化过程,探讨CD34+祖细胞、嗜酸性粒细胞、CCR3/eotaxin与哮喘小鼠肺部炎症的关系。 方法 以C57BL/6小鼠为研究对象,以卵白蛋白作为抗原建立哮喘模型,于最后一次抗原激发后6、12、24、48 h检测BALF、外周血和骨髓中的嗜酸性粒细胞、CD34+祖细胞、eotaxin的变化,检测CD34+祖细胞上CCR3的表达;行肺组织病理切片观察嗜酸性粒细胞浸润;PCR检测肺组织CCR3和eotaxin mRNA水平。 结果 抗原激发后嗜酸性粒细胞数、CD34+细胞数、CD34+/CCR3+细胞数在BALF、外周血、骨髓悬液中有不同程度的增加。模型组BALF的eotaxin水平在抗原激发后6 h与对照组相比有明显增加(P<0.05),并一直持续到24 h。外周血和骨髓悬液中eotaxin水平与对照组相比差异无统计学意义(均P>0.05)。模型组各时间点肺组织eotaxin mRNA和CCR3 mRNA的表达与对照组相比均有明显增加。BALF中嗜酸性粒细胞数分别与骨髓悬液中嗜酸性粒细胞数、CD34+/CCR3+细胞数呈正相关,而与骨髓悬液中CD34+细胞数无明显相关性。 结论 ①哮喘小鼠气道局部嗜酸性粒细胞增多与骨髓CD34+祖细胞上CCR3表达上调有关。②CCR3/eotaxin参与CD34+祖细胞分化和趋化,与哮喘小鼠气道嗜酸性粒细胞浸润密切相关。      

Inducing effects of macrophage stimulating protein on the expansion of early hematopoietic progenitor cells in liquid culture

Background Macrophage stimulating protein （MSP） is produced by human bone marrow endothelial cells. In this study we sought to observe its effects on inducing the expansion of early hematopoietic progenitor cells which were cultured in a liquid culture system in the presence of the combination of stem cell factor （SCF）, interleukin 3 （IL-3）, interleukin 6 （IL-6）, granulocyte macrophage-colony stimulating factor （GM-CSF）, erythropoietin （EPO） （Cys） and MSP or of Cys and bone marrow endothelial cell conditioned medium （EC-CM）. Methods Human bone marrow CD34^＋ cells were separated and cultured in a liquid culture system for 6 days. Granulocyte-macrophage colony forming unit （CFU-GM） and colony forming unit-granulocyte, erythrocyte, macrophage, megakaryocyte （CFU-GEMM） were employed to assay the effects of different treatment on the proliferation of hematopoeitic stem/progenitor cells. The nitroblue tetrazolium （NBT） reductive test and hoechest 33258 staining were employed to reflect the differentiation and apoptosis of the cells respectively. Results MSP inhibited the proliferation of CFU-GM and CFU-GEMM in semi-solid culture and the inhibitory effect on CFU-GEMM was stronger than on CFU-GM. MSP inhibited the differentiation of early hematopoietic progenitor cells induced by hematopoietic stimulators. Bone marrow （BM） CFU-GEMM was 2.3-fold or 1.7-fold increase or significantly decreased in either Cys＋EC-CM, Cys＋MSP or Cys compared with 0 hour control in liquid culture system after 6 days. Conclusion MSP, a hematopoietic inhibitor, inhibits the differentiation of early hematopoietic progenitor cells induced by hematopoietic stimulators and makes the early hematopoietic progenitor cells expand in a liquid culture system.     

Effect of Ligustrazine on the Expression of LFA-1, ICAM-1 Following Bone Marrow Transplantation in Mice

Asablood activatingChinesedrug ,ligustrazinecanimprovethemicroenvironmentofbonemarrow ,increasetheadherentfunctionofstromalcellsandpromotethegrowthofhematopoieticcells[1] .Toin terprettherolesthatligustrazineplaysinbonemar rowhematopoieticreconstitutionan…     

肾母细胞瘤基因衍生肽诱导细胞毒性T淋巴细胞对白血病患者骨髓CD34+细胞的体外杀伤效应

目的 探讨肾母细胞瘤基因(WT1)衍生肽负载树突状细胞(DC)诱导细胞毒性T淋巴细胞(CTL)对白血病CD34+细胞的体外清除效应.方法 合成一段针对HLA-A*0201锚位的WT19聚肽,体外负载来源于HLA-A*0201*健康人的DC后,诱导产生WT1肽特异性CTL(A组),以噻唑盐(MTT)比色法观察其对WT1表达阳性白血病患者(HLA-A* 0201+者3例,HLA-A*0201-者3例)骨髓CD34+细胞、健康人(HLA-A*0201+者2例,HLA-A*0201-者1例)外周血CD34+细胞和白血病NB4、K562及U937细胞株的体外杀伤效应,粒细胞-巨噬细胞系集落形成试验观察其对白血病患者骨髓CD34+细胞和健康人外周血CD34+细胞粒细胞-巨噬细胞系集落形成单位(CFU-GM)形成的影响.设立单独DC诱导CTL(B组)和IL-2诱导T细胞(C组)作为对照.结果 在效靶比为20:1时,A组CTL对3例HLA-A*0201+白血病患者骨髓CD34+细胞和NB4细胞的杀伤活性(分别为55.3%±2.8%,67.1%±3.2%、49.4%±3.8%和55.0%±3.7%)明显高于对3例HLA-A*0201-白血病患者骨髓CD34+细胞、健康人外周血CD34+细胞及K562、U937细胞的杀伤活性(均<20%),并明显高于B组和C组CTL(均P＜0.01).2例HLA-A*0201+白血病患者骨髓CD34+细胞经A组CTL处理后CFU-GM集落相对形成率分别为17.8%±4.0%和20.8%±3.4%,明显低于经B组CTL处理后(分别为88.9%±3.4%和91.8%±5.7%,均P＜0.01);HLA-A*0201-白血病患者骨髓CD34+细胞、健康人外周血CD34+细胞经A组和B组CTL处理后CFU-GM集落相对形成率差异尤统计学意义.结论 WT1肽特异性CTL能够以HLA-1类抗原限制方式杀伤高表达WT1基因的白血病CD34+细胞,且能特异性抑制其CFU-GM集落形成,WT1基因的表达产物可以作为清除白血病CD34+细胞靶点.     

胚胎干细胞来源造血细胞的细胞表型和功能分析

【目的】 分析经卵黄囊(YS)、胎肝(FL)和骨髓来源(BM)的基质细胞条件培养液(SCCM)诱导产生的胚胎干细胞(ES)来源造血细胞的细胞表型与功能差异。【方法】 制备YS-SCCM、 FL-SCCM及BM-SCCM,将3种基质细胞条件培养液分别加入ESE 14.1细胞分化培养体系培养7 d,通过对分化ESE14.1细胞造血发育表面标志FLK-1、Integrin-α4(整联蛋白-α4、Sca-1(干细胞抗原-1)、CD34的检测、体外高增殖潜能集落形成细胞(HPP-CFC)分析及体内脾集落形成单位(CFU-S)检测,评价3种基质细胞条件培养液对ESE14.1细胞体外造血发育的调控作用。【结果】 经FL-SCCM诱导的EB细胞Flk-1、Integrinα4+ 和Sca-1+ 细胞均高于YS-SCCM和BMSC-CM诱导组,分别为3.03%、2.9%和13.74%;经BMSC-CM诱导产生的CD34+ 细胞比例最高,为1.07% 。经FLSC-CM或BMSC-CM诱导产生的造血细胞其HPP-CFC产率明显高于对照组,分别为7.4个/105细胞(P < 0.01)和5.8个/105细胞(P < 0.05);经FLSC-CM或BMSC-CM诱导产生的造血细胞其CFU-S产率亦明显高于对照组,分别为8.5个/5 × 105细胞和6.75个/5 × 105细胞(P < 0.001)。【结论】 YS-SCCM、 FL-SCCM及BM-SCCM均可诱导ESE14.1向造血细胞分化,FL-SCCM和BM-SCCM造血定向诱导效率较高,所产生的细胞具备造血细胞的正常功能,FL-SCCM诱导产生的造血细胞原始程度高于BM-SCCM诱导产生的造血细胞。     

肺动脉高压大鼠骨髓内皮祖细胞功能的研究

目的检测实验性肺动脉高压大鼠骨髓内皮祖细胞（EPC）的功能,探讨肺动脉高压发病机制。方法利用野百合碱诱导大鼠发生肺动脉高压,分离骨髓单个核细胞进行体外诱导培养以获得EPC集落,并对其骨髓内皮祖细胞的集落形成能力、增殖、黏附、迁移能力进行检测。结果骨髓单个核细胞在体外培养下能够获得EPC集落,与对照组比较,肺动脉高压实验组诱导生成的EPC数量少（P〈0.05）,CD34和FLK-1阴性比例下降（分别为19.33%±3.27%vs 31.17%±4.40%和33.67%±3.50%vs 44.50%±3.78%,P均〈0.01）,细胞增殖能力下降（0.43±0.08 vs 0.64±0.07,P〈0.01）,贴壁细胞减少（6.835个±1.605个vs 10.175个±1.945个,P〈0.01）,细胞迁移能力下降（7.83个±1.94个vs 11.83个±2.48个,P〈0.05）。结论肺动脉高压的发生与骨髓内皮祖细胞功能的异常存在明显的相关性。