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…     

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

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

Effects of adenovirus mediated vascular endothelial growth factor gene transfer on reconstitution of hematopoiesis in post-bone marrow transplantation mice

Background Bone marrow transplantation (BMT) conditioning procedure is considered as the cause of damage to bone marrow microvasculature and the delay of hematopoiesis recovery. However, hematopoiesis regulation post BMT by vascular endothelial growth factor (VEGF) has not yet been studied. In this study, adenovirus were used to investigate the effects of VEGF gene transfer on preventing damages to bone marrow microenvironment and its promotion of hematopoiesis in post-BMT mice.Methods Recombinant adenovirus (Ad)-enhanced green fluorescent protein (EGFP)/hVEGF165 was injected via tail vein into BALB/c mice undergoing syngeneic BMT. During the different phases post BMT, the distribution of adenovirus and the plasma levels of hVEGF were measured as well as the numbers of white blood cells (WBC), platelet (PLT) and red blood cells (RBC) in peripheral blood. At the same time, the mice were injected with Chinese ink via tail vein, following which the tibias were separated and were used for analysis of bone marrow microvasculature surface area and cellularity.Results Significant expression of EGFP and hVEGF was observed in multiple organs at different phases post BMT, and the plasma level of hVEGF was up to (866.67±97.13) pg/ml. The recovery of WBC, PLT and RBC of the group treated with recombinant adenovirus Ad-EGFP/hVEGF165 were significantly more rapid than those of other BMT groups （P<0.05, respectively）. At the 20th day post BMT, the percentage of bone marrow microvasculature surface area in group treated with VEGF ［(61.2±4.0)%］ returned to normal level ［(62.0±5.0)%, P>0.05］. The restoration of hematopoiesis was retarded more than that of microvasculature. The cellularity of bone marrow in each group was still lower than that of normal control ［(62.3±4.0)%, P<0.05］ at the 30th day post BMT, but the percentage in group treated with VEGF at the 20th and 30th days post BMT ［(46.5±5.0)% and (55.1±4.5)%］ exceeded those of other BMT groups (P<0.05, respectively).Conclusion VEGF gene transfer mediated by adenovirus may protect the hematopoietic microenvironment to promote the restoration of hematopoiesis in post-BMT mice.     

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…     

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.     

小鼠骨髓移植修复卵巢功能损伤

目的研究骨髓移植对化疗引起的小鼠卵巢功能损伤的修复作用。方法 48只小鼠随机分为3组:正常对照组(A组)、化疗并骨髓移植组(B组)和化疗组(C组)。B、C组小鼠每天给予环磷酰胺150 mg/kg腹腔注射,连续3 d。1周后,对B组小鼠进行骨髓移植。给药起第21、50天分两批处死小鼠,称量卵巢、子宫湿重,观察卵巢组织学变化。用Hoechest33342荧光染料标记骨髓细胞,移植入预先注射环磷酰胺的小鼠体内,2 d后取小鼠卵巢做冰冻病理切片,荧光显微镜下观察。结果给药第21、50天,B组小鼠的卵巢系数、各级卵泡数目明显高于C组而低于A组,均具有统计学意义(P<0.05)。受体鼠的卵巢内发现显示荧光的供体鼠骨髓细胞。结论骨髓移植对化疗药物引起的卵巢功能损伤具有修复作用。     

肝细胞生长因子对骨髓移植小鼠造血重建的促进作用

目的 观察肝细胞生长因子（HGF）对骨髓移植小鼠造血重建的促进作用。方法 用BALB／c小鼠建立同基因骨髓移植模型，随机将BALB／c小鼠分A、B两组，B组移植后叶7d皮下注射HGF，A组皮下注射PBS，移植后1、7、14、21和28d检测小鼠血红蛋白、白细胞、血小板、骨髓单个核细胞及骨髓造血细胞面积。结果 B组血红蛋白在移植后7、14d与同期A组无差异（P〉0．05），B组血红蛋白在移植后21d高于同期A组（P〈0．05）；B组血小板在移植后7、14及21d高于同期A组（P值分别小于0．01、0．01和0．05）：B组白细胞在移植后7、14、21及28d高于同期A组（P值分别小于0．01、0．01、0．05及0．05）；B组骨髓单个核细胞及骨髓造血细胞面积在移植后7、14、21和28d高于同期A组（P值分别小于0．05、0．01、0．01及0．05）。结论 HGF对骨髓移植小鼠造血重建有一定的促进作用。     

Effects of Platelet Factor 4 on Expression of Bone Marrow Heparan Sulfate in Syngenic Bone Marrow Transplantation Mice

Platelet factor 4 ( PF4) is a negativehematopoietic factor.It can inhibit the prolifera-tion of endothelial cells and hematopoietic stem/progenitor cells,particularly megakaryoryocyticcells,reversibly[1] ,inhibit DNA synthesis,blockcell cycle progression during S phase and reducethe sensibility of normal hematopoietic stem/pro-genitor cells,but not some cancer or leukemia celllines,to cytotoxic drugs and ionizing radia-tion[2 - 3] ,and it also can cause a population in-crease of the stem cel…     

单倍相合异基因骨髓移植前后骨髓像分析

目的观察单倍相合异基因骨髓移植前后骨髓像、造血能否重建。方法对22例患者造血干细胞移植前骨髓穿刺抽吸涂片镜检;骨髓移植成功出层流室首次骨髓穿刺抽吸涂片;每月定期复查骨髓穿刺抽吸涂片镜栓。同时观察外周血涂片和骨髓涂片2张或2张以上。结果22例患者单倍相合并基因造血干细胞移植后骨髓像全部达到正常,完全缓解。移植成功随访,其中6例患者随访中骨髓细胞出现少许病态造血现象,2例出现纯红细胞性再生障碍性贫血（PRCA）,余14例骨髓正常。结论单倍相合并基因骨髓移植能够使白血病和重型再障骨髓重建。造血重建恢复期可能继发骨髓增生异常综合症（MDS）和PRCA。     

小鼠不同细胞数骨髓移植后回巢与生存率观察

目的:探讨移植细胞数对小鼠骨髓移植回巢及生存率的影响。方法:采用不同细胞数移植、不同细胞数加不同时问间隔程序移植及流式细胞仪检测等方法,观察不同细胞数骨髓移植后小鼠骨髓内回巢效果和生存情况。结果:不同细胞数骨髓移植中5×105组的回巢细胞数与高细胞数组无差异,回巢率远远高于后者(P<0.01);生存率1×105组为95％,与高细胞数组无差异。不同细胞数加不同时间间隔程序移植以细胞总数在1×105、间隔24 h移植组存活率高,组间差异非常显著(P<0.01)。结论:在一定细胞数量范围内,分次移植可能提高移植干细胞的回巢效率。     

自然杀伤细胞对小鼠异基因骨髓移植中移植物抗宿主病效应的影响

目的:研究自然杀伤细胞(NK)在异基因骨髓移植中对移植物抗宿主病(GVHD)的影响.方法:以近交系小鼠C57BL/6(H-2b)为供鼠、BALB/c(H-2d)为受鼠,在移植物中增加供者的外周T细胞和(或)NK细胞进行异基因骨髓移植,根据临床表现和病理检查,比较不同移植组的存活率和GVHD发生情况.结果:增加1×106供鼠NK细胞的移植组中40%小鼠出现GVHD,增加2×106供鼠NK细胞的移植组中仅30%出现GVHD;与单纯异基因骨髓移植细(80%急性GVHD、20%慢性GVHD)和骨髓 T细胞组(100%急性GVHD)比较,GVHD发生率显著下降(P＜0.01),30、60和120 d存活率显著增高(P＜0.01).结论:在小鼠异基因骨髓移植中,同种异基因反应性NK细胞可抑制GVHD效应,延长存活期.     

骨髓间充质干细胞移植重建极重度放射损伤小鼠造血功能

目的:探讨小鼠骨髓间充质干细胞(mouse bone marrow mesenchymal stem cell,mMSC)单独输注对重度放射损伤小鼠造血功能重建的影响.方法:将体外培养扩增的雄性C57BL/6小鼠骨髓间充质干细胞输注给受致死剂量全身照射(8 Gy)的雌性C57BL/6小鼠,检测移植后外周血象、骨髓有核细胞数、病理变化、粒-单核细胞系祖细胞集落(CFU-GM)计数及性染色体比例,以未输注MSCs的小鼠作对照.结果:对照组小鼠(n=6)在照射后20 d内全部死于造血功能衰竭;MSCs组移植后血象明显下降,但2周后迅速恢复,28 d时恢复到照射前的60%左右,42 d外周血象基本恢复.MSC促进骨髓有核细胞数及CFU-GM快速恢复,有利于骨髓组织学的明显恢复改善.移植后42 d,仍可在受致死量照射的受体鼠内检测到供体MSC,但不能长期植入.结论:小鼠骨髓间充质干细胞单独输注可促进重度放射损伤小鼠的造血功能恢复,其植入时间长短可能与输注的细胞量有关.     

骨髓干细胞移植后mdx鼠抗肌萎缩蛋白的表达

目的研究骨髓干细胞移植治疗Duchenne型肌营养不良鼠(mdx鼠)后抗肌萎缩蛋白表达的动态变化。方法取6~8周龄C57BL/6鼠骨髓干细胞,以2×107个/只干细胞尾静脉移植到经7Gyγ射线预处理后的7~9周龄mdx鼠体内,分别于移植后5、8、12、16周应用免疫荧光、逆转录-聚合酶链反应、Western blot检测腓肠肌抗肌萎缩蛋白的表达情况。结果经γ射线放疗预处理的mdx鼠,在尾静脉移植骨髓干细胞5周后可见抗肌萎缩蛋白少量表达,随移植时间延长表达逐渐增多,16周时肌纤维抗肌萎缩蛋白的表达为12%,并可见肌细胞核中移现象较同龄mdx鼠对照组减少,边居增多。结论同种异体骨髓干细胞移植治疗mdx鼠后,骨髓干细胞通过血液循环趋向病变肌组织,并分化为肌细胞表达抗肌萎缩蛋白。随移植时间延长表达增多,提示骨髓干细胞移植可长久持续参与受损骨骼肌的修复与再生。     

NK细胞对小鼠H-2单倍体相合骨髓移植后GVHD及造血重建的影响

目的:研究自然杀伤(NK)细胞在单倍体相合骨髓移植中对移植物抗宿主病(GVHD)及造血重建的影响. 方法: 以近交系Balb/c(H-2d)×C57BL/6(H-2b)杂交F1代小鼠(H-2d/b)为供鼠、Balb/c(H-2d)为受鼠,在移植物中增加供者的外周T细胞和(或)NK细胞进行半相合骨髓移植,根据临床表现、病理检查及外周血白细胞(WBC)数,比较不同移植组GVHD发生及造血重建情况. 结果: 骨髓 T细胞组中90%的小鼠出现GVHD,外周血WBC平均在移植后4 wk恢复正常;而骨髓 T 低浓度NK细胞组中20%小鼠出现GVHD, 外周血WBC平均在移植后3 wk恢复正常;骨髓 T 高浓度NK细胞组中仅10%出现GVHD,外周血WBC也平均在移植后3 wk恢复正常. 后两组较骨髓 T细胞组GVHD发生率显著下降(P＜0.01),外周血WBC恢复时间显著提前(P＜0.01). 结论:在小鼠单倍体相合骨髓移植中,同种反应性NK细胞可降低GVHD发生率,促进造血重建.     

来氟米特预防小鼠异基因骨髓移植后aGVHD的实验研究

目的 评估来氟米特（LEF）对小鼠异基因骨髓移植（allo-BMT）术后急性移植物抗宿主病（aGVHD）的预防作用.方法 以SPF级雄性C57BL/6（H-2b）为供鼠,雌性BALB/C（H-2d）为受鼠,建立allo-BMT/aGVHD动物模型,并将大鼠随机分为：A组（模型对照组）、B组（环孢菌素A组,剂量为10 mg·kg^-1·d^-1）、C组（LEF组,剂量为10 mg·kg^-1·d^-1）,同时根据临床表现、病理组织学检查、平均生存时间评估各组aGVHD的严重程度.结果 B组和C组受鼠平均生存时间均较A组显著延长（均P〈0.01）,B组和C组间平均生存时间的差异均无统计学意义（均P〉0.05）;B组和C组受鼠的aGVHD临床表现和病理改变均较A组明显减轻.结论 LEF可有效预防小鼠allo-BMT术后出现的aGVHD,并可减轻其症状和病理损害程度,同时显著延长平均生存时间.     

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

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川芎嗪对大剂量化疗小鼠骨髓基质细胞损伤的保护作用

目的：观察川芎嗪对大剂量甲氨喋呤（MTX）化疗所致骨髓基质细胞（BMSC）损伤的保护作用及其量效关系。方法：建立大剂量MTX化疗损伤BMSC的小鼠模型后，按每千克体重腹腔注射川芎嗪0mg（对照组）、20mg（Ⅰ组）、40mg(Ⅱ组），1次／d,第21天采取骨髓细胞进行体外BMSC培养，成纤维细胞集落（CFU－F）计数，BMSC层粘附率测定和BMSC层上造血细胞混合集落培养，结果：BMSC生长，CFU－F计数，BMSC层粘附率，BMSC层上造血细胞混合集落数目均表现为Ⅱ组＞Ⅰ组＞对照组（P＜0．05，P＜0．01）。结论：川芎嗪对大剂量MTX引起的BMSC损伤有明显的保护作用，且呈剂量依赖性。     

营养干预对骨髓移植病人抗氧化力及脂质过氧化的影响

目的探讨外源性补充抗氧化性维生素对骨髓移植病人抗氧化力及脂质过氧化的影响。方法将19名骨髓移植病人随机分为实验和对照组,实验组在预处理前给予维生素C(300 mg/d)和维生素E(600 mg/d),对照组不予补充,其余处理两者相同。测定移植前后两组病人的血浆总抗氧化力、丙二醛、全血谷胱甘肽过氧化物酶及红细胞超氧化物歧化酶水平。结果(1)移植前后实验组血浆总抗氧化力、谷胱甘肽过氧化物酶和红细胞超氧化物歧化酶值一直处于正常或正常值附近,对照组则低于正常值,两组相比有显著差异;(2)移植后实验组丙二醛值很快降至正常,对照组则一直处于较高水平,两组有显著差异。结论移植前外源性补充抗氧化性维生素可提高骨髓移植病人机体的抗氧化力,减少了脂质过氧化物的产生,有效缓解移植前大剂量放、化疗对病人机体造成的高氧化胁迫压力。