骨形态发生蛋白激活自身肌肉间质细胞拯救骨髓衰竭

目的探讨骨形态发生蛋白(BMP)激活自身肌肉间质细胞以拯救骨髓造血衰竭的作用。方法采用5-氟脲嘧啶与白消安合用诱导大鼠致死性再生障碍性贫血模型,于诱导再障前3d肌肉内植入重组人-BMP-2(rh-BMP-2),并以肌肉内植入琼脂为对照。观察血象、病理形态等变化以及两组再障大鼠的死亡率。同时,给正常小鼠大腿肌肉内植入rh-BMP-2,动态观察植入后局部的形态变化、形成骨及骨髓的形态特点,脾集落形成单位以及基质细胞干细胞生长因子(SCF)的表达。结果小鼠在BMP肌肉内植入1周内,在BMP周围有大量间质细胞增殖,随后形成骨及骨髓,其骨髓基质细胞的SCF表达明显高于自体骨髓;BMP植入组的再障大鼠,不仅血象与对照相比有明显改善,而且死亡率也明显下降,有56.3%大鼠存活超过3个月,其血象完全恢复正常,骨髓和脾脏的组织形态学改变也恢复正常。对照大鼠除1只存活超过3个月外(占4.3%),其余均死于急性再生障碍性贫血。结论BMP肌肉内植入可拯救骨髓造血衰竭,其机制可能与BMP诱导成年自体肌肉内存在的干细胞向造血分化有关;本研究结果为应用成体自身干细胞进行细胞治疗提供了新的途径。     

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。     

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.     

无细胞胎肝液对大剂量化疗后小鼠骨髓造血细胞的影响

采用小鼠体内液体扩散盒及脾结节法观察了胎鼠和成年小鼠肝脏无细胞液对大剂量环磷酰胺化疗后小鼠骨髓造血细胞增殖与分化的影响。发现无细胞胎肝液能明显促进受抑制骨髓中多能造血干细胞(CFU—S)的集落形成率,但对扩散盒内粒系祖细胞(CFU—D)的增殖分化无促进作用。成年鼠肝无细胞液则对造血细胞的增殖无刺激活性,不能促进化疗后小鼠骨髓中造血干细胞恢复。认为无细胞胎肝液中存在的造血刺激因子能有效缓解细胞毒剂对骨髓多能造血干细胞的抑制。     

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。     

COLONY GROWTH AND GRANULOPOIETIC EFFECTS OF HUMAN BONE MARROW FIBR.OBL.AST COLONY-FORMING CELLS IN VITRO

Human bone marrow derived fibroblast colony- ;forming cells (F-CFU) and granulocytemacrophage colony-forming cells (GM-CFU) were simultaneously studied by liquid and semi-solid agar culture in 26 normal individuals and 33 hematological disorders iii 13 patients with aplastic anemia, 9 acute non lymphocytic leukemia, 6 acute Iymphocytic leukemia and 5 malignant lymphoma. After 2 weeks of liquid culture, the growth rates and morphological charac teristics of F-CFU from aplastic and leukemic mar rows did not differ markedly from the normal, except that patient groups show a wider range of colony number. The F-CFU incidence in patients with malignant lymphoma was lower than normal. Growth of GM-CFU from patients with aplastic anemia and Ieukemia was very poor. GM-CFU stimulation was not detected iii any supernatants of F-CFU cultures. Confluent mono layers of fibroblast cells grown from normal and aplastic marrow could directly stimulate the growth cf GM-CFU of normal marrow cells and little GM- CFU was observed in areas lacking fibroblast cells. Fibroblast monolayers derived from acute leukemic marrows were poor stimulators of GM-CFU, this abnormality in interactions between stromal cells and hematopoietic progenitors may be important in the pathogenesis and clinical manifestation of leukemia. This study suggests that bone marrow fibroblast cells may directly affect hematopoietic stem cell proliferation and differentiation and supports the hypothesis that marrow fibrosis observed in patients with leukemia is of non-ncoplastic origin and may be a reactive phenomenon. Analysis of the pathoge- nesis of aplastic patients with defective hematopoie tic microenvironments may be possible.     

Panaxdiol Saponins Component Promotes Hematopoiesis and Modulates T Lymphocyte Dysregulation in Aplastic Anemia Model Mice

Objective: To investigate the potential efficacy of panaxadiol saponins component (PDS-C) in the treatment of aplastic anemia (AA) model mice. Methods: Totally 70 mice were divided into 7 groups as follows: normal, model, low-, medium-, high-dose PDS-C (20, 40, 80 mg/kg, namely L-, M-, H-PDS-C), cyclosporine (40 mg/kg), and andriol (25 mg/kg) groups, respectively. An immune-mediated AA mouse model was established in BALB/c mice by exposing to 5.0 Gy total body irradiation at 1.0 Gy/min, and injecting with lymphocytes from DBA mice. On day 4 after establishment of AA model, all drugs were intragastrically administered daily for 15 days, respectively, while the mice in the normal and model groups were administered with saline solution. After treatment, the peripheral blood counts, bone marrow pathological examination, colony forming assay of bone marrow culture, T lymphocyte subpopulation analysis, as well as T-bet, GATA-3 and FoxP3 proteins were detected by flow cytometry and Western blot. Results: The peripheral blood of white blood cell (WBC), platelet, neutrophil counts and hemoglobin (Hb) concentration were significantly decreased in the model group compared with the normal group (all P<0.01). In response to 3 dose PDS-C treatment, the WBC, platelet, neutrophil counts were significantly increased at a dose-dependent manner compared with the model group (all P<0.01). The myelosuppression status of AA was significantly reduced in M-, H-PDS-C groups, and hematopoietic cell quantity of bone marrow was more abundant than the model group. The colony numbers of myeloid, erythroid and megakaryocytic progenitor cells in the model group were less than those of the normal mice in bone marrow culture, while, PDS-C therapy enhanced proliferation of hematopoietic progenitor cells by significantly increasing colony numbers (all P<0.01). Furthermore, PDS-C therapy increased peripheral blood CD3+ and CD3+CD4+ cells and reduced CD3+CD8+ cells (P<0.05 or P<0.01). Meanwhile, PDS-C treatment at medium- and high doses groups also increased CD4+CD25+FoxP3+ cells, downregulated T-bet protein expression, and upregulated GATA-3 and FoxP3 protein expressions in spleen cells (P<0.05). Conclusion: PDS-C possesses dual activities, promoting proliferation hematopoietic progenitor cells and modulating T lymphocyte immune functions in the treatment of AA model mice.     

Stimulating effect of catechin, an active component of Spatholobus suberectus Dunn, on bioactivity of hematopoietic growth factor

Background Hematopoietic growth factor （HGF） is indispensable to hematopoiesis in the body. The proliferation and differentiation of hematopoietic cells must rely on the existence and stimulation of HGF. This study investigated the effect of catechin, an active component extracted from Spatholobus suberectus Dunn （SSD）, on bioactivity of granulocyte-macrophage colony-stimulating activity （GM-CSA）, burst-promoting activity （BPA） and megakaryocyte colony-stimulating activity （MK-CSA） in spleen condition medium （SPCM） of mice to clarify the hematopoietic mechanism of catechin and SSD.
Methods Spleen cells of mice were separated and spleen condition medium （SPCM） was prepared from spleen cell culture. Bone marrow cells of mice were separated and cultured in a culture system including 10% （v/v） SPCM （induced by catechin in vivo or ex vivo） for 6 days. Granulocyte-macrophage colony forming units （CFU-GM）, erythrocyte burst-colony-forming units （BFU-E） and megakaryocyte colony-forming units （CFU-Meg） formation were employed to assay the effects of different treatment on the bioactivity of GM-CSA, BPA and MK-CSA in SPCM.
Results SPCM induced by 100 mg/L catechin ex vivo could promote the growth of CFU-GM, BFU-E and CFU-Meg, which indicated that catechin could stimulate the production of GM-CSA, BPA and MK-CSA in SPCM. SPCM prepared at the fourth day of spleen cell culture showed the best stimulating activity. The bioactivity of GM-CSA, BPA and MK-CSA in the SPCM prepared after intraperitoneally injecting catechin into mice was also increased. The number of CFU-GM, BFU-E and CFU-Meg gradually increased as the dose of catechin increased and the time of administration prolonged. CFU-GM, BFU-E and CFU-Meg of the high-dose catechin group were significantly higher than those of the control group （P〈0.01） and reached the maximum at the seventh day after administration.
Conclusions This study suggests that catechin extracted from the active acetic ether part of Spatholo     

The effects of total saponins of panax ginseng on hematopoietic progenitor cells in healthy humans and aplastic anemia patients

Ginseng is said to have beneficial effects on anemia. The proliferation effects of total saponins of Panax ginseng (TSPG) on hematopoietic progenitor cell in healthy individuals and 29 patients with aplastic anemia (AA) were observed through bone marrow cultures of burst forming unit-erythroid (BFU-E), colony forming unit-erythroid (CFU-E) and colony forming unit-granulocyte/macrophage (CFU-GM) in vitro compared with methyltestosterone (MT). The results suggest TSPG might prompt the proliferation of normal progenitor cells at a concentration of 20 μg/ml. The numbers of BFU-E,CFU-E and CFU-GM increased by 37.8±2.9%, 31.4±2.9% and 33.3±4.0% respectively overthe controls; furthermore TSPG was still useful to BFU-E, CFU-E growth without Epo in vitro, although the colony numbers were much lower. Otherwise MT was useless to CFU-GM. Of the 29 patients with AA, 14 who responded to MT showed sensitivity to TSPG in marrow culture (the rising rate of colony formation exceeded 30%), but immune-mediated AA (patient’s peripheral blood mononucleated cell suppressed normal hematopoiesis) and stem cell decreased AA (few of colonies were formed) showed almost no expression for TSPG activity because of the immunological suppression system and the absence of progenitors.     

黄芪注射液对贫血小鼠巨核系造血的作用及其机理的研究

OBJECTIVE: To assess the effect of Astragalus membranaceus injection (AMI) on megakaryocyte hematopoiesis in anemic mice and explore its mechanism. METHODS: Anemic models of mice were randomly divided into two groups: treatment group and anemic control group. Intraperitoneal doses of AMI (20 mg/(ml.20 g) q.d x 6) were given to the treatment group, and equal doses of physiological saline were given to the anemic control group. On days 8, 11 and 14 after treatment, blood platelet and bone marrow cells were determined, and the numbers of CFU-Meg (colony forming unit-megakaryocyte) and Meg-CSA (megakaryocyte colony-stimulating activity) were determined by using technique of hematopoietic progenitor cells culture in vitro. RESULTS: Serum Meg-CSA of the treatment group was significantly higher than that of the anemic control group. The abovementioned indices of the treatment group recovered to normal by day 11, which was markedly earlier than the day of recovery observed in the anemic control group. CONCLUSION: AMI can increase serum Meg-CSA of anemic mice and accelerate the recovery of megekaryocyte hematopoiesis after bone marrow suppression.     

骨形态发生蛋白诱导髓外造血的实验研究

目的:探讨骨形态发生蛋白(BMP)诱导髓外造血以拯救骨髓造血衰竭的作用。方法:采用60Co-γ+氯霉素(CH)+环磷酰胺(CY)的方法诱导小鼠致死性再生障碍性贫血模型,实验组于诱导再障前6d肌肉内植入重组人-BMP-4(rh-BMP-4),对照组小鼠肌肉内植入琼脂。观察血象、病理形态等变化以及两组再障小鼠的死亡率。同时,给正常小鼠大腿肌肉内植入rh-BMP-4,动态观察植入后局部的形态变化、形成骨及骨髓的形态特点,脾集落形成单位(CFU-S)以及基质细胞干细胞生长因子(SCF)的表达。结果:正常小鼠在BMP肌肉内植入1周内,在BMP周围有大量间质细胞增殖,2周后才出现软骨化骨和骨小结形成,其骨髓基质细胞的SCF表达明显高于自体骨髓;BMP植入组的再障小鼠,不仅血象与对照组相比有明显改善,而且死亡率也明显下降,有46.7%存活超过3个月,其血象完全恢复正常,骨髓和脾脏的组织形态学改变也恢复正常。对照小鼠除1只存活超过3个月外(占6.7%),其余均死于急性再生障碍性贫血。结论:肌肉内植入BMP-4可诱导髓外造血,其机制可能与BMP-4诱导成年自体肌肉内存在的干细胞向造血分化有关,这一结果将为干细胞治疗提供了一条新的思路。     

骨髓基质细胞支持CD34+细胞扩增的实验研究

目的体外模拟造血微环境，观察CD34^ 细胞在骨髓基质细胞支持下的扩增效应。方法将免疫磁珠阳性选择分选的骨髓CD34^ 细胞接种于构建的基质细胞层培养，通过骨髓基质细胞支持CD34^ 扩增的细胞总数及集落形成细胞数(Colonyforming Cell。CFC)的变化，评价骨髓基质细胞支持CD34^ 细胞扩增的功能。结果 扩增后细胞总数及CFC数分别增加，表示骨髓基质细胞能很好地支持CD34^ 细胞扩增。结论 体外证实了骨髓基质细胞在造血干细胞更新、增殖中具有重要作用。     

血虚证小鼠模型骨髓细胞红系祖细胞、粒单系祖细胞变化规律及中药复方对其的影响

目的：研究血虚证小鼠模型骨髓造血祖细胞体外增殖水平及变化规律,以及中药复方地甘口服液,红霖四物口服液对其的影响。方法：以Balb/c小鼠为实验对象,分别采用免疫介导法和综合放血法制作血虚证模型。采集其骨髓有核细胞进行培养;并分组灌服地甘口服液,红霖四物口服液,比较观察各组小鼠骨髓细胞体外培养CFU－E,BFU－E,CFU－GM集落的形成情况。结果：两种血虚证小鼠模型的CFU－E,BFU－E,CFU－GM集落形成较空白对照组显下降,尤以前为甚;两种中药复方均可促进模型小鼠骨髓细胞的增殖,但对于免疫介导血虚证模型,两治疗组的效果存在一定差异。结论：血虚证小鼠模型骨髓造血干（红9系,粒单系）细胞的体外增殖较正常小鼠明显下降,而地甘口服液,红霖四物口服液可以促进骨髓CFU－E,BFU－E,CFU－GM集落的形成。     

干细胞因子与白介素-3联合对小鼠骨髓细胞体外扩增作用及其造血重建的影响

采用集落培养的方法研究了干细胞因子（ Ｓ Ｃ Ｆ） 联合白介素 ３（ Ｉ Ｌ ３） 对小鼠５ 氟尿嘧啶（５ Ｆ Ｕ ） 处理３ ｄ 的骨髓细胞（ｄ３ ５ Ｆ Ｕ Ｂ Ｍ Ｃ） 的体外扩增作用及其对致死量照射小鼠造血重建功能的影响。 Ｓ Ｃ Ｆ 单独应用使体系中粒 巨噬细胞集落形成单位（ Ｇ Ｍ Ｃ Ｆ Ｕ） 和高增殖潜能集落形成单位（ Ｈ Ｐ Ｐ Ｃ Ｆ Ｕ） 分别增加了（２４ ．２ ±６ ．１） 和（１ ．５ ±０ ．３） 倍（ Ｐ＜ ０ ．０５） 。 Ｓ Ｃ Ｆ 与 Ｉ Ｌ ３ 联合应用时 Ｇ Ｍ Ｃ Ｆ Ｕ 和 Ｈ Ｐ Ｐ Ｃ Ｆ Ｕ 分别是新鲜ｄ３ ５ Ｆ Ｕ Ｂ Ｍ Ｃ 的（３０ ．５ ±９ ．５） 和（１３ ．６ ±４ ．５）倍（ 与单用 Ｓ Ｃ Ｆ 组比较 Ｐ 分别大于和小于０ ．０５） 。小鼠骨髓移植实验结果表明： Ｓ Ｃ Ｆ、 Ｓ Ｃ Ｆ＋ Ｉ Ｌ ３ 体外扩增的ｄ３ ５ Ｆ Ｕ Ｂ Ｍ Ｃ 移植给经致死量照射的受体小鼠后,外周血细胞计数恢复时间比输新鲜ｄ３ ５ Ｆ Ｕ Ｂ Ｍ Ｃ 组缩短１ ～５ ｄ , Ｓ Ｃ Ｆ＋ Ｉ Ｌ ３组不仅血细胞计数恢复最快,并使移植所需的骨髓细胞数减少９０ ％ 以上。结果提示： Ｓ Ｃ Ｆ 单用对骨髓细胞有扩增作用,并且与 Ｉ Ｌ ３ 有协同作用;体外扩增的骨髓?     

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…     

有机锗的抗放射作用

ICR小鼠受致死量射线照射以后,接受有机锗治疗14天.剂量为500mg/kg的小鼠30天活存率比对照组提高37.5％;当Ce—132剂量为500mg/kg和1000mg/kg时,其内源性脾结节的数量明显地高于对照组。实验结果表明,Ce—132具有一定的治疗急性骨髓型放射病的作用和刺激内源性造血干细胞的增殖与分化作用。     

益血胶囊对环磷酰胺致血虚证小鼠骨髓造血的影响

[目的]考察益血胶囊对环磷酰胺(cycloph osphamide,CTX)致血虚证小鼠骨髓造血和外周血象的影响.[方法]70只昆明小鼠随机分5组,分别为正常对照组、模型对照组、益血胶囊低、中、高剂量组.剂量分别设置为5 g·kg-1,10 g·kg-1,20 g·kg-1.采用ip CTX 250mg/(kg·d)腹腔注射制成血虚证模型小鼠,模型制作后立即口服灌胃,1次/d,连续10d,在0,1,3,5,7,10,14,17,21d检测外周血象,在第10d进行集落培养.[结果]益血胶囊对CTX 致血虚证小鼠骨髓中红系祖细胞(CFU-E、BFU-E)、粒系祖细胞(CFU-GM)数量的回升有明显促进作用(P＜0.05),益血胶囊对CTX致血虚证小鼠外周血白细胞和血小板的恢复有明显的促进作用(P＜0.05).[结论]益血胶囊可促进CTX致血虚证小鼠的骨髓造血,进而升高外周血中的白细胞和和血小板,这可能是益血胶囊用于临床肿瘤患者化疗的作用机制.     

气血康口服液抗辐射的实验研究

采用内源性脾集落形成法，骨髓有核细胞计数及脾脏称重法观察了气血康口服液对辐射小鼠造血功能的影响。研究结果表明，经气血康处理的小鼠ＣＦＵ－Ｓ、骨髓有核细胞数及脾重等项指标均明显明显高于阴性对照组和阳性对照组，提示气血康口服液在辐射损伤紧急状态下，不仅可促进髓内造血细胞生成，而且对动物体内髓外造血也有显著促进作用。其抗辐射的确切机理有待于进一步研究。     

小鼠颌下腺组织培养上清液影响红系造血的实验研究

为了研究颌下腺是否合成造血因子以及合成的造血因子的可能种类 ,进一步研究颌下腺对血发生的调控 ,采用造血祖细胞体外培养和流式细胞术检测细胞表面标记物。结果显示 ,小鼠颌下腺组织培养上清液(SGCM)在体外培养中 ,对早期和晚期红系造血祖细胞有促进增殖和分化的作用 ,实验组集落数远远高于阴性对照组 (P<0 .0 5 ) ;晚期红系祖细胞 (CFU- E)培养体系雄性小鼠颌下腺组织培养上清液组集落数明显高于雌性组 (P<0 .0 5 ) ,早期红系祖细胞 (BFU- E)培养体系没有明显差异 (P>0 .0 5 ) ,贫血模型小鼠颌下腺促进 BFU- E和 CFU- E增殖和分化的活性高于正常小鼠 (P<0 .0 5 ) ,IL- 3与 SGCM有协同刺激作用 ,说明小鼠颌下腺确能合成和分泌促进红系造血的细胞因子 ,雄性小鼠颌下腺促进 CFU- E增殖和分化细胞因子的活性高于雌性小鼠。本实验为进一步研究小鼠颌下腺合成造血因子和其与造血调控的关系提供了实验依据。     

Effect of Spirulina Platensis Polysaccharide on Hematopoietic Recovery and Related Cytokines in Mice with Transplanted Tumor Treated by Chemotherapy

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