人工合成成骨生长肽对正常小鼠造血的促进作用和体外造血活性测定

目的　通过观察sOGP在体内的促造血活性和体外促进造血祖细胞增殖的作用 ,初步探索sOGP促造血活性的作用环节及机制。方法　正常小鼠连续 14d皮下注射sOGP ,观察外周血象、骨髓有核细胞数及骨髓有核细胞分类计数的变化 ;又应用离体骨髓细胞培养方法观察sOGP对正常小鼠和人骨髓粒系祖细胞集落形成的影响。结果　sOGP能升高正常小鼠骨髓有核细胞数和外周血白细胞数分别为2 0 %和 40 % ,红细胞和血小板也有增加趋势 ,骨髓细胞增生较空白对照组活跃 ,粒系增生较明显。体外CFU G集落培养实验进一步证实了sOGP具有直接刺激小鼠和人骨髓粒系祖细胞增殖的作用 ,1× 10 -14 mol·L-1浓度时已有明显的效果 ,sOGP和阳性对照药rhG CSF在相近浓度时效果相接近。结论　sOGP可能具有促进小鼠骨髓全系细胞增生的作用 ,以促进粒系造血为主 ,其机制可能是直接作用于造血干 /祖细胞或改善造血微环境促进造血 ,提示sOGP在促进放 /化疗患者造血功能恢复等方面潜在的应用价值     

肉苁蓉总苷对~(60)Coγ射线照射小鼠造血系统损伤保护作用的研究

目的　研究肉苁蓉总苷 (GCs )对60 Coγ射线照射小鼠造血系统损伤的辐射防护作用 ,探讨其作用机制。方法　连续动态观察GCs对小鼠经60 Coγ射线照射后d 3,d 7,d14,d 2 1外周血血红蛋白、白细胞、网织红细胞、骨髓有核细胞数、脾集落数 (CFU S)以及骨髓细胞硒 谷胱甘肽过氧化物酶 (Se GSH Px)活性和丙二醛 (MDA)含量的影响。结果　辐射损伤后小鼠外周血白细胞、网织红细胞、骨髓有核细胞数、CFU S明显减少 ,骨髓细胞及脾Se GSH Px活性降低 ,MDA含量明显增加。GCs可促进上述各造血指标的恢复 ,提高Se GSH Px活性 ,降低MDA含量。结论　GCs对受照小鼠造血系统具有保护作用 ,其机制可能与抗氧化作用有关。     

Positive effects of indomethacin on restoration of splenic nucleated cell populations in mice given sublethal irradiation

Splenic cellularity during the recovery phase after 400-R irradiation was evaluated in mice, in which the level of prostaglandin was regulated by indomethacin and exogenous prostaglandin E2, following sublethal irradiation. Two weeks after irradiation, administration to these mice of indomethacin, an inhibitor of prostaglandin synthesis, augmented the recovery of all nucleated spleen cell populations, whereas the thymus was drastically depopulated. This treatment had little effect upon the total number of bone marrow cells but inversed the ratio of PNA+/PNA- cells. Cell transfer experiments using heavily irradiated mice as recipients showed that the stem cell proliferation was positively affected by indomethacin treatment in the bone marrow rather than in the spleen. These results suggest that cell migration from primary lymphoid organs, particularly from the bone marrow to the spleen, is regulated by a prostaglandin-mediated system and that a prostaglandin E2 synthesis inhibition would have a positive effect on the restoration of peripheral nucleated cells following irradiation.     

Limits of stimulation of proliferation and differentiation of bone marrow cells of mice treated with swainsonine

The limits of stimulation of the immunomodulatory alkaloid swainsonine (8alphabeta-indolizidine-1alpha,2alpha,8beta-triol) were studied in inbred C57BL/6 mice for potential support of intense high dose cancer chemotherapy and/or radiation because of its attractive pharmacologic profile on the hematopoietic system. Specifically, the effects of swainsonine on bone marrow cellularity and on in vitro progenitor cell proliferation to total colony forming units (CFU) and differentiation to different lineages were studied as a function of number of days post drug administration. The lineages evaluated were colony forming units-granulocyte-macrophage (CFU-GM), erythroid-burst forming units (BFU-e) and CFU-granulocyte-erythrocyte-monocyte-megakaryocyte (CFU-GEMM or CFU-Mix). Groups of mice were treated with swainsonine or plain vehicle, phosphate buffered saline for 10 consecutive days. The effects of these agents on the hematopoietic system were studied up to 60 days following their discontinuation. The magnitude of the effects of swainsonine on bone marrow system gradually declined with increasing duration of days following its discontinuation. Nevertheless, its residual stimulatory effects on bone marrow cellularity, total CFU, CFU-GM, BFU-e and CFU-Mix continued to be significant (P<0.0001) up to 45, 50, 50, 55 and 50 days, respectively, compared to those of diluent buffer or untreated controls. Since cancer chemotherapeutic agents or radiation are normally given in schedules and/or cycles, these results strongly suggest that swainsonine effects are sustained long enough to potentially support and facilitate hematopoietic recovery during anti-cancer cytotoxic treatment.     

六种重组细胞因子对人骨髓造血细胞的体外扩增效应

目的 :观察造血生长因子对骨髓CD34 +细胞的扩增效应及扩增细胞的分化特性。方法 :采用重组人白细胞介素 (rhIL) 1 +rhIL 3+rhIL6 +重组人粒细胞集落刺激因子 +重组人粒 巨噬细胞集落刺激因子和重组人干细胞因子对人骨髓单个核细胞 (MNC)进行刺激 ,动态观察了经上述细胞因子作用后MNC的增殖效应及液态扩增培养后粒 巨噬集落形成率 (CFU GM ) ,利用流式细胞技术 (FACS)分析了扩增前后CD34 +细胞及其亚群的动态变化。结果 :经上述 6种细胞因子作用 2 4d ,骨髓MNC总数是原代MNC的 38.33倍 ,液态扩增 1 0d后 ,CFU GM的数量达高峰 ,是原代MNC的 1 9.0 4倍 ,2 4d时CFU GM总数降至 3.94倍 ,且集落明显小于扩增初期。扩增至 6～ 1 0d ,CD34 +细胞总数是原代MNC的 1 36 .40～ 90 .40倍 ,1 9d时降为 6 1 .40倍。结论 :外源性造血生长因子对CD34 +骨髓细胞具有较强的扩增效应 ,选择适宜的扩增时间是保证骨髓移植后尽快重建造血功能的重要环节。     

仿刺参糖胺聚糖对骨髓抑制贫血小鼠外周血及骨髓细胞周期的影响△

目的：观察仿刺参糖胺聚糖(HGAG)对环磷酰胺诱导的骨髓抑制贫血小鼠外周血及骨髓细胞周期的影响。方法：将40只雄性昆明种小鼠随机分为正常组、模型组、HGAG治疗组(1,5,15mg.kg_-1),每组8只。除正常组外其余各组腹腔注射环磷酰胺100mg.kg_-1制备贫血小鼠模型,每天1次连续3天,并于造模当日起,HGAG给药组腹腔注射不同浓度的HGAG,正常组和模型组腹腔注射生理盐水,每天1次连续7天。用全自动血细胞分析仪、白细胞计数法、流式细胞术分别检测HGAG对贫血小鼠外周血、骨髓有核细胞数和骨髓细胞周期的影响。结果：HGAG低、中、高剂量组能显著升高外周血WBC和HGB,且高剂量组还能明显升高外周血RBC和PLT。HGAG三个剂量组能显著增加小鼠骨髓有核细胞数和脾脏指数,而且能够促进骨髓G₀/G₁期细胞向S期及S期细胞向G₂/M期细胞的转化,显著升高增殖指数。结论：HGAG能够解除细胞周期阻滞,促进骨髓造血细胞的增殖,增加外周血细胞、骨髓有核细胞的数量和升高脾脏指数,从而促进骨髓抑制贫血小鼠造血功能的恢复     

Accelerated recovery from cyclophosphamide-induced leukopenia in mice administered a Japanese ethical herbal drug, Hochu-ekki-to

The effect of a Japanese ethical herbal drug, Hochu-ekki-to (HOT), on recovery from leukopenia induced by cyclophosphamide (CY) was investigated. Daily oral administration of 1000 mg/kg HOT into CY-treated mice significantly prevented decrease of leukocyte numbers in the peripheral blood and accelerated recovery from leukopenia. Ginsenoside Rgl extracted from Ginseng radix, a major herb of HOT, was one of the active ingredients. HOT increased numbers of neutrophils and monocytes in the peripheral blood compared with CY-treated control. Moreover, HOT augmented the resistance against Pseudomonas aeruginosa infection. The number of colony-forming units in the spleen (CFU-S) also increased in HOT-treated mice. The frequencies of IL-3-, GM-CSF- and IFN-gamma-producing cells increased in the spleen, bone marrow, liver and IEL on HOT treatment, and HOT clearly augmented the expressions of IL-3, GM-CSF and IFN-gamma mRNA in the spleen, bone marrow, liver and IEL except IL-3 and IFN-gamma mRNA in the IEL. These results suggest that HOT enhances the production of hematopoietic lymphokines, stimulates the proliferation of hematopoietic progenitor cells and consequently accelerates recovery from leukopenia in CY-treated mice. Additionally, IFN-gamma which HOT-augmented the production may contribute the protective effect against the bacterial infection by activating of phagocyte cells.     

螺旋藻多糖对小鼠移植性肿瘤化疗后造血恢复及相关细胞因子的影响

目的：研究螺旋藻多糖（polysaccharide from spirulina platensis,PSP)对小鼠移植性肿瘤化疗后造血恢复及相关细胞因子的影响。方法：通过右前肢皮下接种肿瘤细胞形成小鼠移植性实体瘤，给予PSP边疆灌胃10d,d4给予环磷酰胺（CTX）腹腔注射连续3d。于d11，分别了外周细胞，骨髓有核细胞及CFU－S计数，用紫外分光光度计检测骨髓DNA含量，用双抗体夹心ELISA法检测血清中IL-1,IL-3,GM-CSF,TNF含量。结果：CTX可造成明显骨髓抑制，而PSP则提升了外周血细胞，增加了骨髓中有核细胞计数和DNA含量以及促进了CFU－S形成。此外，PSP还增加了血清中IL－1，IL－3，GM－CSF，TNF含量。结论：PSP可能通过促进内源性细胞因子的分泌来实现其促进小鼠移植性肿瘤化疗损伤后的造血恢复。     

Acute toxicity of benzene inhalation to hemopoietic precursor cells

When BDF₁ mice were submitted to inhalation of benzene vapors (4680 ppm) for 8 hr, a significant depletion in bone marrow colony forming cells (CFC) was observed in cultures in vitro assayed 1 day after drug inhalation; however, femoral CFC returned to steady state values by Day 7. The return of femoral CFC values was not due to an increase in bone marrow cellularity. Although spleen cellularity increased significantly by Days 4 and 7, the splenic CFC value was not significantly different from controls. This acute toxicity to femoral CFC was enhanced by multiple sessions of benzene inhalation over a period of 3 and 3.5 days. After three and one-half 8-hr sessions of benzene vapor inhalation (4680 ppm), CFC content was dramatically reduced (13% of controls), in addition to significantly lowered values for body and spleen weights and bone marrow cellularity. Splenic colony forming unit (CFU) formation in irradiated mice which received bone marrow cells from benzene-treated mice corresponded well with in vitro femoral CFC formation from mice treated with the identical multiple benzene inhalation regimen as the donor mice; CFU and CFC production was reduced to the same extent, about 40–45% of controls. In summary, precursor cells of the hemic cell renewal system are sensitive to benzene inhalation in mice.     

Effect of SCG, 1,3-beta-D-glucan from Sparassis crispa on the hematopoietic response in cyclophosphamide induced leukopenic mice

Sparassis crispa Fr. is an edible mushroom recently cultivable in Japan. It contains a remarkably high content of 6-branched 1,3-beta-D-glucan showing antitumor activity. Using ion-exchange chromatography, a purified beta-glucan preparation, SCG, was prepared. In this study, we examined the hematopoietic response by SCG in cyclophosphamide (CY)-induced leukopenic mice. SCG enhanced the hematopoietic response in CY induced leukopenic mice by intraperitoneal routes over a wide range of concentrations. SCG enhanced the hematopoietic response in CY-treated mice by prior or post administration. Analyzing the leukocyte population by flow cytometry, monocytes and granulocytes in the peritoneal cavity, liver, spleen and bone marrow (BM) recovered faster than in the control group. The ratio of natural killer cells and gammadelta T cells in the liver, spleen and peritoneal cavity was also increased. In contrast, CD4+ CD8+ cells in the thymus were temporarily significantly decreased by the administration of SCG. Interleukin-6 (IL-6) production of CY+SCG-treated peritoneal exdated cells (PECs), spleen cells and bone marrow cells (BMCs) were higher than that of the CY-treated group. By in vitro culture of CY-treated PEC and spleen cells, IL-6 production was enhanced by the addition of SCG. These facts suggested the possibility that IL-6 might be a key cytokine for the enhanced hematopoietic response by SCG.     

Swainsonine stimulates bone marrow cell proliferation and differentiation in different strains of inbred mice.

The immunomodulatory alkaloid swainsonine (8alphabeta-indolizidine-1alpha,2alpha,8beta-triol) has potential for overcoming the bone marrow suppressive effects of cancer chemotherapeutic drugs and radiation. The effect of swainsonine on bone marrow cellularity was evaluated in four different strains (C57BL/6; C3H-HEN; Balb/C and DBA-2 mice) of inbred mice subjected to multiple doses of the alkaloid. Swainsonine treatment stimulated bone marrow cell proliferation in all strains of mice. Examination of the peripheral blood did not reveal any increase in total leukocyte count. In vitro assessment of total colony-forming unit (CFU) capacity of bone marrow cells showed a two- to eight-fold increase in swainsonine treated mice of different strains compared to their corresponding controls given sham injections of physiological saline. Swainsonine induced increase in CFU capacity of bone marrow cells should find clinical application in cancer treatment with chemotherapeutic agents and radiation.     

莪术提取物ERC61对化疗小鼠造血功能的影响

目的:观察莪术提取物ERC61对环磷酰胺(CTX)抑制小鼠造血功能的影响并确定其最佳给药方式。方法:实验小鼠ipCTX(100mg.kg-1.d-1),连续给药3d,复制白细胞减少症模型;将ERC61按照单纯预防、单纯治疗和预防+治疗3种给药方式ip给药,常规方法计数小鼠白细胞数、骨髓有核细胞数,并测定脾指数。结果:ERC61对CTX抑制小鼠的白细胞数、骨髓有核细胞数、脾指数均有不同程度的改善。结论:ERC61可改善CTX抑制小鼠的造血功能,其中预防+治疗的给药方式效果最好。     

Isopropyl methanesulfonate is a potent red cell hematotoxicant

Mice given 10-weekly injections (i.m.) of 20 mumol of isopropyl methanesulfonate (IPMS) in trioctanoin exhibited depressed hematopoiesis as manifested by reduced peripheral blood counts and by reduced thymic and bone marrow cellularities. The treatments were particularly toxic to erythroid populations and ultimately induced signs of splenic erythropoiesis. Bone marrow cellularity recovered during the latter stages of the treatments indicating that subpopulations of hematopoietic cells were resistant to the treatments. This recovery of bone marrow cellularity may bear on the recently discovered leukemogenic potential of IPMS.     

芪术口服液对环磷酰胺所致小鼠骨髓造血功能损伤的保护作用

目的:观察芪术口服液对环磷酰胺所致小鼠骨髓造血功能损伤的保护作用.方法:对小鼠腹腔注射100 mg/kg环磷酰胺后,观察芪术口服液对小鼠外周血象、骨髓有核细胞和胸腺、脾脏指数的影响.结果:芪术口服液能升高受损小鼠白细胞、红细胞、血小板、血红蛋白和骨髓有核细胞数量,并能使小鼠胸腺、脾脏指数升高.结论:芪术口服液对环磷酰胺引起的小鼠骨髓造血功能损伤有一定的保护作用.     

Modulation of myelopoiesis by CSF or CSF-inducing biological response modifiers

We have compared the effects on number and function of bone marrow progenitors and peripheral effector cells of the myelomonocytic lineage of treatment with the 2-cyanaziridine compounds Azimexone and BM 41.332 to those of maleic anhydride divinyl ether copolymer (MVE-2) or granulocyte-macrophage colony stimulation factor (GM-CSF). Within a few hours after i.p. injection of either Azimexone or BM 41.332, there was a dose-dependent increase in serum CSF levels, CSF secretion by mononuclear bone marrow cells (BMC) and macrophages (M phi), which was followed by an increase in granulocyte-M phi committed stem cells (GM-CFU-C), nucleated BMC, and peripheral blood leukocytes. Optimal effects occurred 3 days after 50 mg/kg Azimexone or 25 mg/kg BM 41.332. Three i.p. injections of 50 mg/kg Azimexone into mice pretreated with cyclophosphamide (CY) (150 mg/kg) were able to significantly restore suppressed bone marrow cellularity (GM-CFU-C and nucleated BMC). Azimexone also increased the number of peripheral M phi in normal or CY-treated mice, without inducing detectable tumoricidal activity. These M phi, however, retained their capacity to become fully activated (cytotoxic) by appropriate activation signals such as IFN or LPS. Analogous to the 2-cyanaziridines. MVE-2 (at 25 mg/kg) had similar stimulatory effects on myeloid functions in normal mice. MVE-2 induced, in addition, a significant augmentation of cytoxicity by both M phi and NK cells. In contrast, single or multiple injections of semipurified GM-CSF into normal mice (1000 U or 5000 U per mouse) failed to detectably stimulate myelopoietic growth and differentiation. 2-cyanaziridine compounds thus offer the potential of selectively augmenting growth and differentiation of myelomonocytic cells in normal and bone marrow-depressed mice without appreciably affecting their immunological status.     

刺梨汁对骨髓抑制小鼠造血功能的调控作用

目的探讨刺梨汁对环磷酰胺合并60Co致骨髓抑制小鼠造血功能的影响,研究其促进血细胞上调的作用机制。方法双抗体夹心法检测外周血常规、骨髓基质中红细胞生成素（EPO）、血小板生成素（TPO）、粒细胞生长因子（GCSF）的量。结果刺梨汁能明显改善骨髓抑制小鼠外周血常规,能有效平衡微环境中TPO的量,改善EPO的表达,对骨髓抑制小鼠骨髓细胞上清中GCSF有明显的正调控作用。结论刺梨汁能提高骨髓抑制小鼠外周血血细胞和骨髓有核细胞计数,可促进骨髓抑制小鼠骨髓细胞从G0/G1期进入增殖周期,并能有效调节骨髓微环境中TPO、EPO、GCSF的表达,从而促进骨髓抑制小鼠的造血功能,可作为一种治疗贫血的辅助保健药物。     

黄芪多糖对丝裂霉素C(MMC)致骨髓抑制小鼠骨髓及脾脏造血祖细胞的生成作用的影响

目的　研究黄芪多糖 (APS)对MMC致骨髓抑制小鼠的骨髓和脾造血祖细胞生长的影响。方法　d 0腹腔注射丝裂霉素C(MMC) 7mg·kg- 1 ,APS皮下注射 1 0 0mg·kg- 1·d- 1 ,给药方案分 3种 :① 0～ 4d ,共 5d ;② 0～ 1 1d ,共 1 2d ;③ 1 2d给药 ,3wk内给完 ,用造血细胞集落培养法观察APS的药理作用。结果　在d 3时 ,APS治疗组对MMC致骨髓抑制小鼠骨髓CFU C数目增加了 3倍 ,分别为 1 870±40 /股骨和 62 4 0± 1 1 0 /股骨 ,从d 3～d 1 8,APS治疗组的骨髓CFU C均高于模型组。在给MMC后d 1 4之前APS对脾CFU C的生长没有影响 ,APS在d 1 4和d 1 8时APS有刺激脾造血祖细胞增殖作用 (分别高 3倍和 2倍 )。结论　APS对MMC引起骨髓抑制小鼠有促进骨髓和脾脏造血祖细胞的增殖和成熟的作用     

Dose-dependent responses of murine pluripotent stem cells and myeloid and erythroid progenitor cells following administration of the immunomodulating agent glucan

The hemopoietic effects produced by six different doses of a commercially available glucan preparation were investigated. C3H/HeN mice were intravenously injected with either 0.1, 0.4, 0.8, 1.2, 1.6, or 2.0 mg of glucan. Five days later, total nucleated cellularity, pluripotent stem cells (CFU-s), granulocyte-macrophage colony-forming cells (GM-CFC), macrophage colony-forming cells (M-CFC), and erythroid burst- and colony-forming cells (BFU-e and CFU-e) were assayed in the bone marrow and spleen. Neither femoral cellularity nor GM-CFC content was altered by any of the glucan doses tested. By contrast, bone marrow CFU-s content increased and CFU-e and BFU-e contents decreased with increasing glucan doses. In the spleen, all aspects of hemopoiesis (i.e., cellularity, CFU-s, GM-CFC, M-CFC, and CFU-e) increased after glucan administration. The degree of splenic enhancement was directly correlated with the dose of glucan administered.     

益血生胶囊对小鼠辐射损伤的保护作用

目的:研究益血生胶囊对辐射损伤的治疗作用.方法:以X射线照射小鼠全身引起辐射损伤,观察益血生胶囊对辐射小鼠白细胞、骨髓有核细胞、胸腺、脾脏的影响.结果:益血生胶囊能升高受照鼠白细胞、骨髓有核细胞数,并使受照鼠胸腺、脾脏重量回升.结论:益血生胶囊对X射线所致辐射损伤有较好保护作用.