Enhancing Effects of Ligustrazine on Expression of CD31 and Hematopoietic Reconstitution in Syngenic Bone Marrow Transplantation of Mice

Summary： The effect of ligustrazine on the expression of CD31 in syngenic bone marrow transplantation （BMT） mice was studied. Fifty-six Balb/c mice were divided into 3 groups： normal control group. BMT control group, and ligustrazine treated group. Syngenic BMT mouse models were established according to the literatures. In BMT control group and the ligustrazine treated group, the mice were given respecxively orally 0.2 mL saline and 2 mg ligustrazine twice a day. On the 7th, 14th, and 21st day after BMT, the mice were killed. The expression of CD31 on the surface of bone marrow nuclear cells （BMNC） was detected by flow cytometry. Peripheral blood leukocytes, platelets and BMNC were counted. Histological observation of bone marrow was made. The results showed thai in ligustrazine treated group the peripheral blood leukocylcs, platelets and BMNC counts, and the expression of CD31 on the day 7, 14, 21 after BMT were higher than in BMTcontrol group （P〈0.01 or P〈0.05）. In conclusion, ligustrazine could obviously enhance the CD31 expression on the surface of BMNC after syngcnic BMT in mice, which may be one of the mecha- nisms underlying the ligustrazine accelerating hematopoietic reconstitution in syngenic BMT.     

Effects of Ligustrazine on Expression of Bone Marrow Heparan Sulfates in Syngeneic Bone Marrow Transplantation Mice

Hematopoietic reconstitution after bone marrowtransplantation (BMT ) is a complicated process,which concerns notonly with the quality and quanti-ty of hematopoietic stem cells(HSC) ,but also withthe recovery of hematopoietic inducing microenviron-ment (HIM) .The contactof HSC and HIM,whichregulates the proliferation,differentiation and self-maintenance of HSC,is critical for hematopoietic re-constitution.Heparan sulfates (HS) are a kind of gly-cosaminoglycans macromolecules composed of…     

Study on the effect of ligustrazine on hematopoietic reconstitution in bone marrow transplantation mice

Hematopoietic reconstitution after bone marrowtransplantation is a complicated process CXCchemokines and their receptors play important rolesin this process. One of these receptors, CXCR4 expresses on hematopoietic stem cells, and is now takenas a "homing factor" in hematopoietic reconstitu--'tionll' 2]. Ligustrazine can protect the bone marrowmicroenvironment after irradiation or immune damage[', 4]. We established allogenic bone marrow transplantation mice models and explored the effects …     

Effect of ligustrazine on hematopoiesis in bone marrow transplantation mice

Hematopoietic reconstitution after bone marrowtransplantation is a complicated process,whichconcerns with the quality and quantity ofhematopoietic stem cell (HSC) ,and also with therecovery of hematopoietic inducing microenvironment(HIM) .The contact of HSC and HIM is critical forhematopoietic reconstitution.1　MATERIALSAND METHODS1 .1　Materials1 .1 .1　 Animals　　 BALB/c mice of both sexes(clean class,close copulatory series (H- 2 d,MLSb) )were 8— 1 0 weeks old,and 1 7— 2 …     

川芎嗪对骨髓移植小鼠造血细胞SDF-1表达水平的影响

为探讨川芎嗪对骨髓移植(BMT)后造血重建过程中基质细胞来源因子(SDF-1)的作用,建立了典型同基因BMT小鼠模型,胃饲川芎嗪注射液每次2 mg,2次/d,分别于BMT后第7、14 d计数外周血细胞骨髓单核细胞(BMNC)并测定骨髓组织切片中SDF-1的表达.结果表明川芎嗪组BMT后第7、14 d外周血细胞计数、BMNC计数及骨髓组织切片中SDF-1表达水平均高于BMT组.提示川芎嗪能在骨髓移植造血重建早期促进骨髓基质细胞表达SDF-1,可能是促进BMT后造血干细胞(HSC)回髓、加速造血重建的机制之一.     

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…     

Effects of Ligustrazine on Hematopoiesis in the Early Phase of Bone Marrow Transplantation Mice

the National NaturalScience Foundation(Serial No.3 9870 92 6)sacrificed on the1 st,3rd,5 th,7th,1 0 th,1 5 th,and 2 1 st day after BMT.The marrow from fe-murs was flushed into RPMI- 1 640 medium. BM-NCs from each femur were counted.1 .2 .7Carbon- Mold Preparation of Blood Vessels　　 A colloidal carbon solution( prepared Chineseink,0 .2 ml,2 0 mg/ml in saline) was injected1 -2 min before sacrifice on the1 st,3rd,5 th,7th,1 0 th,1 5 th,2 1 st day after BMT.One femur ofeach mouse was r…     

骨髓腔内移植对异基因骨髓移植小鼠造血的影响

目的:探讨骨髓腔内骨髓移植(Intrabonemarrowbonemarrowtransplantation,IBMBMT)对异基因骨髓移植小鼠造血功能的影响。方法:将供鼠C57BL/6(H2b)骨髓细胞经微量注射器直接注入到经致死量照射后的受鼠BalB/c(H2d)胫骨骨髓腔。在骨髓移植后1,7,14d分别检测受鼠外周血红细胞、白细胞、血小板;骨髓移植后7,14d分别检测受鼠骨髓中的CFUS。结果:IBMBMT组小鼠移植后7,14d的外周血红细胞、白细胞、血小板、骨髓脾集落形成单位(CFUS)均显著高于传统骨髓移植组(IVBMT)(P<0.01)。结论:IBMBMT较IVBMT更能促进异基因骨髓移植后造血功能重建。     

Study on the effects of Guiqi Oral Liquid in promoting recovery of hematopoiesis in acute irradiation injured mice

Objective: To explore the effects and possible mechanisms of Guiqi Oral Liquid (GQOL) on the recovery of hematopoiesis in acute irradiation injured mice.Methods: The acute irradiation injured mice were randomly divided into 2 groups: the treated group and the control group, and also a normal control group was set up with 6 mice in it receiving no treatment. After the mice in the former two groups were irradiated by 6.0 Gy⁶⁰Coγ-ray, every one of them was given 0. 4 ml GQOL or saline in equal volume through a gastric tube twice a day for 14 days. On the 4th, 8th and 14th day after irradiation, the bone marrow mononuclear cells (BMMNC) and megakaryocytes in bone marrow tissues of the mice were counted, the proportion of hematopoietic tissues (by area) was measured, and the expression of adhesion molecules, CD44 and CD54, in bone marrow were estimated by immunochemistry. The colony forming unit of spleen (CFU-S) in the mice were counted on the 8th day after irradiation.Results: On the 4th, 8th, 14th day after irradiation, the count of BMMNC and megakaryocyte, and the proportion of hematopoietic tissues in the treated group were higher than those in the control group (P<0.01 orP<0.05). CD44 and CD54 expression in the treated group were higher than those in the control group on the 4th and 8th day (P<0.01), but near normal on the 14th day (P<0.01). On the 8th day, CFU-S count in the treated group was higher than that in the control group (P<0.01).Conclusion: GQOL can regulate the expression of adhesion molecules, CD44 and CD54, in the bone marrow of the acute irradiation injured mice, which may be one of the mechanisms of GQOL in accelerating the early phase hematopoiesis recovery of mice. Supported by the National Natural Science Foundation of China (No. 39870926)     

Study on the Protective Effects of Compound Blood-activating Soup on Bone Marrow Hematopoietic Cells in Acute Radiation Injured Mice

ThisprojectwassupportedbyNationalScienceFoundationofChina(No.39670911)andTraditionalChineseMedicineSectionofthePublicHealthDepartmentofHubeiProvince.Ithasbeenknownthattheprominentmanifestationofbonemarrowintheearlyperiodofacuteradiationwasthathematopoieticcellsdecreasedrapidly,venoussinusexpanded,becominghyperemic,andbloodflowwasinstasisll].Onthebasisofthetheoryofblood-activatingandstasiseliminatingofChinesetraditionalmedicine,weusedablood--activatingandstasis--eliminatingChinesedrug,aco…     

组胺H_2-受体激动剂和H_1-受体激动剂对多能造血干细胞(CFU-S)细胞周期影响的拮抗作用

本实验显示组胺H_2受体激动剂4-甲基组胺可促使更多的多能造血干细胞进入细胞周期,而组胺H_1受体激动剂2-甲基组胺则能拮抗4-甲基组胺的这一作用。4-甲基组胺与2-甲基组胺对造血干细胞周期状态的影响之间又呈现相互竞争现象。文中讨论了上述实验事实的理论意义。     

外源性mFasL-cDNA基因干预后骨髓造血细胞功能的实验研究

目的:探讨FasL基因转移后造血细胞活性是否到影响,骨髓移植(BMT)后受者骨髓造血功能是否受改变。方法:FasL-cDNA经鉴定后,常规收集BalB/c小鼠和BAC小鼠股骨和胫骨骨髓单个核细胞(BMMNC),采用脂质体转移法将FasL基因转入BalB/c小鼠BMMNC,然后按1∶0.625体外与BAC鼠BMMNC混合培养,6d后实验组(第3组)尾静脉输注给经致死量照射的BalB/c小鼠,同时设立:第1组(空白对照组即未移植细胞);第2组(未转染基因BalB/c鼠BMMNC+BAC鼠BMMNC);第4组(同基因BMT组)。观察移植后受者鼠造血功能及细胞来源,移植物抗宿主病(GVHD)及生存率。结果:BMT后10,20d,第4组白细胞及血小板计数明显高于第2、3组(P<0.01);第2、3组间差异不显著(P>0.05);30d,各组间均无显著性差异(P>0.05);第1组小鼠相继于1周内死亡,死亡后取脾观察未见脾结节形成。第2组、第3组存活的11只雌性BalB/c小鼠BMMNC中,Y染色体出现率为(79.35±6.77)%。第2组、第3组、第4组2月生存率分别为30%、80%、100%。第3组生存率明显高于第2组(P<0.01),与第4组无显著差异(P>0.05)。第3组少数及第2组中大部分鼠移植30d出现GVHD表现,两组死亡鼠及第2组存活鼠组织因子改变符合Ⅱ-Ⅲ度GVHD病理变化,第3组存活7只鼠中有Ⅰ度GVHD病理变化。结论:转基因自体造血细胞与异基因供体BMMNC混合培养后移植,能使受者获给供体来源的造血重建,并明显降低GVHD发生。     

Effects of Ligustrazine on Bone Marrow Microvessel Systemin the Early Period of Acute Radiation Injury in Mice

Ithasbeenprovedbymanystudlesthatthedifferent1ationanddevelopmentofthehematopoieticcellsdependonthesup-portandregulationofbonemarr0wmi-croenvironment,inwhichthevascularsys-temisanimp0rtantpart.Kn0speetalre-portedthatafterhigh-doseirradiationde-stroyedthehematopoieticcellsandthemi-crovesselsysteminbonemarrow,there-coveryofvascularsystemwasessentialforthehematopoiesisrehabilitatioin[l.2].There-fore,itisofgreatsignificancetostudytheeffects0fLigustrazine,akind0fs0called"blood-activatingandstasis-el…     

NK细胞受体与MHC I类分子匹配程度对骨髓造血重建影响的初步研究

目的：探讨供鼠NK细胞受体与受鼠MHCⅠ类分子匹配程度对骨髓移植小鼠造血重建的影响。方法分离30只C57BL/c小鼠的骨髓细胞和NK细胞,输给经过预处理的BALB/c和CB6F1小鼠,受鼠分为单纯辐照组、输入骨髓细胞组、输入骨髓细胞和NK细胞组,6只/组,观察不同时间点血常规、生存时间、病理组织学等指标变化,进行组间比较。结果输入骨髓细胞、输入骨髓细胞和NK细胞组小鼠均长期存活,未观察到GVHD病理学改变。移植后第14天时,输入骨髓细胞组和输入骨髓细胞和NK细胞组的BALB/c小鼠（MHCⅠ类分子全不相合）外周血WBC分别为（1.35＆#177;0.33）＆#215;109/L和（1.80＆#177;0.18）＆#215;109/L,差异有统计学意义,血小板计数分别为（79＆#177;19）＆#215;109/L和（117＆#177;13）＆#215;109/L,差异有统计学意义;输入骨髓细胞组和输入骨髓细胞和NK细胞组的CB6F1小鼠（MHCⅠ类分子半相合）外周血 WBC 分别为（1.52＆#177;0.26）＆#215;109/L 和（1.85＆#177;0.34）＆#215;109/L,差异无统计学意义,血小板计数分别为（90＆#177;12）＆#215;109/L 和（113＆#177;15）＆#215;109/L,差异具有统计学意义;BALB/c小鼠白细胞的改善略优于CB6F1小鼠。结论异源反应性NK细胞可以促进骨髓移植小鼠造血功能的恢复,并且这种作用似与NK细胞受体与受鼠MHC I类分子的匹配程度相关。     

mFasL基因转移诱导异基因骨髓移植受者获得免疫赦免的实验研究

目的 :通过导入外源FasL cDNA ,诱导异基因骨髓移植物中针对受者MHC的淋巴细胞凋亡 ,从而防止移植物抗宿主病 (GVHD)的发生。方法 :pBillneo mFasL质粒经酶切鉴定后 ,采用脂质体转移法导入Balb c鼠骨髓单个核细胞 (BMNC) ,体外与BAC鼠骨髓移植物混合培养后 ,输注给经致死量照射的Balb c小鼠 ,通过观察受者存活率、GVHD的表现、肠、肝、皮肤病理组织学检查 ,以观察受者鼠GVHD的发生情况 ,同时观察受者鼠的脾结节以了解其造血功能重建。受者鼠骨髓细胞Y染色体分析 ,以查明供者来源的造血干细胞是否植入。结果 :pBillneo mFasL内mFasL cDNA酶切鉴定提示可以按顺序合成出功能蛋白质 ,采用脂质体法传导入BMNC后 ,Westen Blot显示出抗FasL抗体识别的条带。转基因BMNC与BAC鼠脾单个核细胞 (SMNC)混合培养后 ,经流式细胞仪检测 ,SMNC凋亡率明显高于对照组 [(14 .76 %± 0 .36 ) %vs(1.8± 0 .4 2 ) % ,t=3.0 1,P <0 .0 0 1]。骨髓移植 (BMT)后 ,受者GVHD的发生率实验组明显低于对照组 (4 0 %vs10 0 % ,χ2 =6 .0 0P <0 .0 5 )。Y染色体分析 ,提示有供者来源的造血干细胞植入并增殖活化。移植 10d后受者即有脾结节形成 ,2 0d达正常水平。结论 :采用脂质体转移法 ,能将mFasL cDNA转入鼠BMNC并能有效地表达 ,转基因     

Effect of Ligustrazine on Bone Marrow Microenvironment and Signal Transducti on Path in Irradiation Injured Mice

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The Effects of Ligustrazine on the Expression of bFGF and bFGFR in Bone Marrow in Radiation Injured Mice

Thebonemarrowhematopoiesisinductivemi croenvironmentismadeupofstromalcells ,extracel lularmatrixandmicrangiumsystem .Micrangiumsystemisanimportant partinthebonemarrowhematopoiesisinductivemicroenvironment .Bonemarrowisoneoforganswhicharesensitivetoradia tion .Basicfibroblastgrowthfactor (bFGF)isasin glechain peptide[1] ,whichincludes 15 4aminoacids[2 ] .bFGFbindstoitsspecificreceptor(bFGFR) ,whichstimulatestheproliferationandim migrationofendothelialcells .Itplaysanimportantroleintheembryo…     

扶正养营颗粒对免疫介导再生障碍性贫血小鼠骨髓增殖及bcl-2表达的影响

目的:观察扶正养营颗粒对免疫介导再生障碍性贫血小鼠骨髓增殖及bcl-2表达的影响.方法:BALB/c小鼠40只随机分为正常组、模型对照组、环孢霉素A对照组、扶正养营颗粒实验组,记数外周血象、骨髓有核细胞,测定骨髓造血组织容量及bcl-2表达.结果:与正常组比较,模型组小鼠外周血象记数、骨髓有核细胞数、造血组织容量及bcl-2表达显著下降,差异有统计学意义(P＜0.05);与模型组比较,环孢霉素A组、扶正养营颗粒组外周血象记数、骨髓有核细胞数、造血组织容量及bcl-2表达显著增加,差异有统计学意义(P＜0.05).结论:扶正养营颗粒可能是通过上调拮抗凋亡因子bcl-2的表达,改善骨髓增殖,发挥其治疗再生障碍性贫血的作用.     

当归注射液对环磷酰胺损伤小鼠骨髓细胞组织形态及超微结构的保护作用

目的探讨当归注射液对环磷酰胺损伤小鼠骨髓细胞组织形态及超微结构的保护作用。方法将30只Balb/c小鼠随机分为正常组、模型组和治疗组。于第7天断颈处死小鼠,取出尺骨和股骨,计数骨髓单个核细胞,并行尺骨组织学切片和超微结构观察。结果与正常组比较,模型组骨髓单个核细胞明显减少(P<0.01);而治疗组与模型组比较,骨髓单个核细胞明显增多(P<0.01)。骨髓超微结构观察显示模型组线粒体和内质网数量减少且明显受损,血窦减少、窦壁断裂。治疗组线粒体、内质网和血窦数量增加,形态基本正常。结论当归注射液能通过促进骨髓造血细胞增生,改善骨髓微环境而对受环磷酰胺损伤的小鼠进行保护。     

单倍体相合骨髓移植白血病患者造血重建的临床分析和实践

目的：观察单倍体相合骨髓移植治疗白血病的造血重建效果。方法选择36例白血病患者作为研究对象,均采取单倍体相合骨髓移植方案进行治疗,对造血重建指标进行观察评估。结果所有患者均在接受骨髓移植后的3~7 d 内白细胞下降至0,中性粒细胞〉0.5×109/ L 和血小板〉20.0×109/ L 的时间分别为（17.5±0.9） d 和（21.5±0.7）d。所有患者3系造血均得到重建。结论骨髓供者通过人粒细胞集落刺激因子预处理后进行骨髓采集,联合多种免疫制剂,能够有效抑制 HLA 多样性屏障,提高白血病患者的造血重建效果,值得重视。