组胺受体激动剂和拮抗剂对粒单系祖细胞增殖的影响

用体外琼脂培养法研究了2-噻唑乙胺、dimaprit及甲氰咪胍对小鼠CFU—GM产率的影响。结果表明:10~(-8)M～10~(-4)M的2-噻唑乙胺不影响CFU—GM产率,而10~(-8)M的dimaprit即可明显增加CFU—GM产率。此作用随剂量的递增而加强,至10~(-5)M时,增至最大值,此后不再增高。单用甲氰咪胍不影响CFU—GM产率,但伍用dimaprit时,甲氰咪胍阻断dimaprit的上述作用;二者对CFU—GM产率的影响呈现竞争现象。分析了该结果的理论意义和临床意义。     

组胺和甲氰咪胍对粒-单系祖细胞的影响

用体外琼脂培养法研究组胺和甲氰咪胍对小鼠粒-单系祖细胞的影响,结果表明,10~(-9)M～10~(-5)M 组胺和10~(-8)M～10~(-4)M 甲氰咪胍分别作用时,对粒-单系祖细胞的产率没有影响,用甲氰咪胍阻断 H_2-受体后,组胺可增加粒-单系祖细胞的产率和自杀率。对此结果的理论意义和临床意义进行了讨论。     

In vitro inhibitory effect of CD_8~ cells from patients with aplastic anemia on normal CFU-GM growth was blocked by cimetidine

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姜黄素对小鼠CFU—GM及WEHI—3B细胞增殖的影响

用体外半固体集落培养方法，观察不同浓度姜黄素对ＢＡＬＢ／ｃ小鼠骨髓粒－巨噬系祖细胞及骨髓单核系白血病干细胞增殖的影响。结果显示：姜黄素呈剂量依赖性抑制ＣＦＵ－ＧＭ和ＷＥＨＩ－３Ｂ细胞增殖，其半抑制剂量分别为１．０３６＊１０＾－５ｍｏｌ．Ｌ＾－１和１．２２０＊１０＾－６ｍｏｌ．Ｌ＾－１。表明姜黄素对ＷＥＨＩ－３Ｂ白血病细胞的抑制作用强于对ＣＦＵ－ＧＭ的作用。     

The murine submandibular gland conditioned medium induce the development of CFU-GM and CFU-meg]

OBJECTIVE: Our purpose is to determine whether murine submandibular gland produce hematopoietic growth factors or not and which hematopoietic growth factor are produced by murine submandibular gland. METHODS: By using the techniques of culture of hematopoietic progenitor cells(CFU-GM, CFU-Meg) ex vivo, flow-cytometry, the effects of murine submandibular gland conditioned medium(SGCM) on the development of CFU-GM and CFU-Meg in mice was studied. RESULTS: SGCM can stimulate the development of CFU-GM and CFU-Meg (without any exogeneous HGF), almost no colony in control (without any exogeneous HGF and SGCM); the promoting activity of male SGCM is higher than female SGCM's in augmenting colony formation of CFU-Meg (P < 0.05), there is no difference of CFU-GM(P > 0.05). The stimulating activity of anemic mice SGCM is higher normal mice SGCM's (P < 0.05). IL-3 can collaborate with female SGCM to stimulate proliferation of CFU-Meg. SGCM can promote bone morrow cell entiring the active proliferative phase(S/G2). CONCLUSION: Submandibular gland can stimulate the development of CFU-GM and CFU-Meg by secreting hematopoietic growth factor-like activities. Our study provide the experimental evidences for clarifying the relationship between submandibular gland and hematopoiesis.     

Effects of Weiganfi on the Hemopoietic Function of the Myelosuppressed Anemic Mice

To study the effect of Weiganli （维肝力） on bone marrow hemopoiesis. Methods： The effects of Weiganli on the peripheral blood picture and the number of bone marrow nucleated cells （BMCs） were observed in myelosuppressed anemic model mice, and the effects of Weiganli on the growth of colony forming unit-granulocyte macrophage （CFU-GM）, colony forming unit-erythroid （CFU-E）, burst forming unit-erythroid （BFU-E）, colony forming unit-megkaryocyte （CFU-Meg） were investigated by in vitro cell culture technique. The hemopoietic stem cells （HSCs, c-kit＋） in bone marrow were double stained with fluorescent antibody PE-C-Kit and FITC-CD45, and the HSCs （c-kit＋） were counted by flow cytometer with CD45/SSC （side scatter） gating. Results： Peripheral blood cell counts and the number of BMCs were significantly improved after Weiganli administration; and bone marrow hemopoietic stem/progenitor cells were significantly increased. Conclusion： Weiganli can effectively promote the recovery of hemopoietic function in the mvelosuooressed anemic mice.     

雌激素受体激动剂对人脐血内皮祖细胞增殖和迁移的影响

目的:观察选择性雌激素受体(ER)激动剂PPT(α亚型激动剂)和DPN(β亚型激动剂)对人脐带血内皮祖细胞(endothelial progenitor cells EPCs)增殖和迁移的影响,并与17β-雌二醇(17β-estradiol,E2)的作用进行比较。方法:采用密度梯度离心法分离获得脐带血单个核细胞,硫氰酸荧光素标记荆豆凝集素I(FITC-UEA-I)、DiI标记的乙酰化低密度脂蛋白(DiI-ac-LDL)双染及CD133免疫荧光鉴定EPCs。培养的EPCs分别加入不同浓度的E2、PPT和DPN(分别为1×10-10mol/L、1×10-9mol/L、1×10-8mol/L、1×10-7mol/L)进行细胞增殖实验分析(WST)、显微镜下EPCs计数及Transwell技术测定对EPCs迁移的影响。结果:E2及PPT呈浓度依赖性促进EPCs的增殖和迁移,PPT刺激作用等同于E2,DPN无刺激作用。结论:雌激素α亚型激动剂PPT明显刺激内皮祖细胞的增殖和迁移,其作用与17β-雌二醇相同。     

Rhodaminel23介导的光动力学疗法对小鼠淋巴细胞增殖及骨髓干祖细胞集落形成的影响

目的　探讨Rhodamine12 3(Rh12 3)介导的光动力学疗法对小鼠淋巴细胞增殖及骨髓干祖细胞集落形成的影响。方法　以C5 7B/ 6小鼠为供鼠 ,BALB/c小鼠为受鼠 ,脾脏淋巴细胞混合培养 ,比较单纯Rh12 3、单纯激光照射、光动力学疗法对小鼠混合淋巴细胞增殖 (MTT法 )及骨髓粒单核巨噬细胞集落形成 (CFU -GM)的影响 (甲基纤维素培养基法 ) ;流式细胞仪检测混合淋巴细胞中CD3 CD6 9 阳性率。结果　Rh12 3组及单纯激光组对细胞增殖无显著影响 ,光动力学组 2 0mW/cm2 以下剂量激光对细胞增殖无明显影响 ,30mW/cm2 以上明显抑制细胞增殖 ;30mW/cm2 以下剂量对CFU -GM集落无明显影响。光动力学疗法后细胞培养 2 4h ,CD3 CD6 9 表达明显下降。结论　光动力学疗法在 30mW/cm2 激光照射时可抑制淋巴细胞增殖 ,降低早期活化T细胞表达 ,但不影响骨髓CFU -GM集落形成     

组胺受体激动剂及其拮抗剂对红系祖细胞产率的影响

研究组胺及其拮抗剂对早期和晚期红系祖细胞(BFU-E 和CFU-E)产率的影响。结果表明,一定浓度的组胺和2—噻唑乙胺对CFU-E和BFU-E的产率均产生抑制效应,而4—甲基组胺则无此作用;事先用甲苯苄二胺阻断H_1-受体,能减弱组胺对CFU-E和BFU-E的抑制效应;相同条件下用甲氰咪胍阴断H_2-受体无此效应。提示组胺对红系祖细胞的抑制效应系通过组胺H_1—受体所介导。     

组胺对N-甲基-D-天门冬氨酸诱发的神经元损伤的改善作用

目的:在细胞水平探讨组胺对N-甲基-D-天门冬氨酸 (NMDA) 诱发的神经元损伤的改善作用及初步机制.方法:采用原代新生大鼠皮层培养的方法,以形态学和二苯基四氮唑溴盐染色为指标观察神经元的损伤和药物的改善作用.结果:组胺在10-4、10-6、10-7、10-8 mol/L浓度时能显著改善NMDA 50 μmol/L作用3 h引起的神经元死亡,并在10-4和10-7 mol/L浓度时分别呈现保护双峰.组胺10-7 mol/L的改善作用只被组胺H2受体西米替丁逆转,而组胺10-4 mol/L的改善作用只被H1受体吡拉明逆转.结论:组胺能减弱NMDA诱发的神经元兴奋性死亡,其保护作用在低浓度时可能与H2受体有关,在高浓度时可能与H1受体有关.     

川芎嗪对辐射致血虚证小鼠骨髓细胞蛋白质表达的影响

目的:观察川芎嗪对辐射致血虚证小鼠骨髓细胞蛋白质表达的影响,探讨川芎嗪补血作用的分子机制。方法:采用3.5 Gy60Coγ射线全身1次性照射,制备小鼠血虚证模型;骨髓集落细胞培养,观察并计数粒系-巨噬细胞集落生成单位(CFU-GM)、爆增型红细胞集落生成单位(BFU-E)、红细胞集落生成单位(CFU-E)、混合集落生成单位(CFU-mix)集落数;利用蛋白质组学寻找差异表达蛋白质。结果:辐射后小鼠骨髓CFU-GM、BFU-E、CFU-E、CFU-mix集落数明显减少,川芎嗪能使其显著回升并部分逆转辐射后蛋白质表达的变化,使小鼠骨髓细胞5种蛋白质表达回升,5种回落。结论:川芎嗪可促进辐射致血虚证小鼠骨髓造血祖细胞增殖,调节多种骨髓细胞蛋白质的表达,后者可能是川芎嗪促进造血细胞生长和增殖的重要机制之一。     

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.     

组胺对β淀粉样蛋白诱导PC12细胞阿尔茨海默病体外模型的保护作用

目的:观察组胺对β淀粉样蛋白1-42(Aβ42)诱导大鼠肾上腺嗜铬瘤(PC12)细胞损伤的改善作用及其相关的受体亚型。方法:运用PC12细胞,采用Aβ42构建阿尔茨海默病体外模型,以形态学和四甲基偶氮唑盐比色法为指标,观察细胞损伤和药物的改善作用。结果:Aβ42浓度依赖性诱导细胞损伤。组胺在10-7、10-6mol/L浓度时能显著改善Aβ42(5μmol/L)作用24h引起的细胞损伤,10-6mol/L保护作用最大。组胺10-6mol/L的保护作用能被组胺H2受体拮抗剂zolantidine,H3受体拮抗剂clobenpropit所逆转,但不能被H1受体拮抗剂苯海拉明所拮抗。结论:组胺能减弱Aβ42诱发的细胞损伤,可能与组胺H2,H3受体有关。     

Immunotoxin depletion of T cells and its effect on hematopoietic progenitor cells in human cord blood

Objective　To study the selective toxicity of immunotoxin (IT) on T cells in cord blood and simultaneously determine its effect on hematopoietic progenitor cells. Methods　The percentage of CD5 and CD8 T cell subsets in cord blood (CB) and bone marrow (BM) as well as peripheral blood (PB) was measured by immunoenzymatic labeling of monoclonal antibodies using immune complexes of alkaline phosphatase and monoclonal anti-alkaline phosphatase (APAAP complexes). One-way mixed lymphocyte cultures (MLC) were performed to compare the proliferative response of CB with that of PB. The proliferative capability of cord blood T cells and T lymphocyte transformation capacity were evaluated in the presence of anti-CD8 or anti-CD5 immunotoxin by one-way MLC and colorimetric MTT (tetrazolium) assay, respectively. The effect of IT on the growth of hematopoietic progenitor cell of colony forming unit-granulocyte and macrophage (CFU-GM), burst forming unit-erythroid(BFU-E), multipotential hemotapoietic progenitors (CFU-Mix) from CB were estimated by colony-forming assays. Results　A certain proportion of CD5 and CD8 T cells existed in CB. The alloproliferative capacity of CB was similar to that of PB. CD5: Ricin at a dosage of 1×10-10-1×10-8 mmol/L and CD8: Ricin concentration in the range of 1×10-9-1×10-8 mmol/L effectively decreased both the proliferative capability of T cells in MLC during CB and T cell transformation. Over the dosage of 1×10-10-1×10-9 mmol/L, both kinds of IT didn’t obviously affect the growth of hematopoietic progenitor cells. Conclusion　CD5: Ricin and CD8: Ricin may effectively deplete T cells and may not significantly inhibit the function of hemaptopoietic cells at a specific dosage.     

A new 2-aminosteroid induces cellular differentiation and upregulates the expression of MafB and Egr-1 genes respectively in HL-60 and K562 leukemia cells

Background In previous work, we suggested that some 2-aminosteroids inhibited proliferation and induced differentiation of both human and murine leukemia cells. Here, we reported the actions of another new 2-aminosteroid designated as H89712 on human leukemia cells. Methods Cell colony counting and MTT assay were used to determine proliferation. Cell morphology, histochemical staining, UV detection and cytometry were used to determine differentiation. RT-PCR was used to detect gene expression. Standard statistical method was used to analyze data. Results H89712 inhibited proliferation of HL-60 leukemia ceils and the inhibition percentage in MTT assay was 18% at the dose of 10^-8 mol/L and 65% at the dose of 10^-5 mol/L, respectively. The inhibition for HL-60 in colony assay was 23% at the dose of 10^-8 mol/L and 96% at the dose of 10^-5 tool/L, respectively. H89712 also induced HL-60 cells toward macrophage-like differentiation. It was verified by flow cytometry that the percentage of positive CD14 expression in differentiated HL-60 cells was about 9 times higher than that of the control at thedose of 10^-8 mol/L and 20 times higher than that of the control at the dose of 10^-5 mol/L respectively, and this action involved upregulation of MafB gene in HL-60 leukemia cells. On the other hand, H89712 inhibited proliferation of K562 leukemia cells and the inhibition of K562 leukemia cells in MTT assay was shown by 34% at the dose of 10^-8 mol/L and 88% at the dose of 10^-5 mol/L respectively. The inhibition of K562 leukemia cells in colony assay was 53% at the dose of 10^-8 mol/L and 100% at the dose of 10^-5 mol/L respectively. H89712 also induced K562 cells toward erythroid-like differentiation and it was verified by flow cytometry that the percentage of positive CD71 expression in differentiated K562 cells was about 9 times higher than that of thecontrol at the dose of 10^-8 mol/L and 16 times higher than that of the control at the dose of 10^-5 mol/L respectively. This action was related to upregulation of Egr-1 gene in K562 leukemia cells. Conclusions Our results showed the important roles played by MafB in macrophage differentiation and Egr-1 in erythroid differentiation of human myeloid leukemia cells.     

胆碱能受体激动剂和拮抗剂对体外CFU-GM增殖和分化的影响

用体外琼脂培养法研究胆碱能受体激动剂和拮抗剂对粒、单造血祖细胞(CFU-GM)增殖和分化的影响。结果显示,10~(-10)-10~(-6)氨甲酰胆碱均能增加CFU-GM的产率,阿托品可以阻断这种效应;氨甲酰胆碱亦能增加CFU*GM的自杀率及培养基中粒细胞集落的比例。文中讨论了CFU-GM上胆碱能受体的功能特征。     

儿童再生障碍性贫血骨髓祖细胞的测定和意义

目的通过对再生障碍性贫血 (AA)患儿红系 (BFU E、CFU E)、粒单核系 (CFU GM)及巨核系 (CFU Meg)祖细胞培养 ,了解骨髓祖细胞增殖情况。方法抽取 4 7例AA患儿骨髓分别作四类祖细胞培养 ,在 7、14d测定其集落数。结果 4 7例AA患儿中 ,四类集落数均值与对照组比较均有显著下降 (P <0 .0 5 ,P <0 .0 1) ,且四类集落数均值在AA三种类型中 ,基本上随病情加重均值逐步下降 ,其中四类集落数低于对照组下限的患儿分别为BFU E 36 .17%、CFU E 85 .11%、CFU GM 74 .4 7%、CFU Meg 91.4 9 %。BFU E、CFU GM和CFU Meg随着患儿疾病的加重 ,低于对照组下限的患儿百分比同步上升 ,且三者的表现与外周血三系表现一致 ,而CFU E却未见上述现象。结论在部分AA患儿中存在着四类祖细胞培养集落数的下降 ,且基本上与患儿病情成正比 ;BFU E的变化较CFU E更能代表患儿骨髓红系祖细胞的变化。     

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。     

生长抑素对白血病患者骨髓细胞CD34抗原表达的影响

目的研究生长抑素八肽类似物奥曲肽(Octreotide,SMS;商品名善得定)对白血病患者骨髓单个核细胞CD34抗原表达的影响,探讨生长抑素对白血病细胞的调控作用.方法采用细胞培养、免疫细胞化学技术及图像分析系统,随机计数CD34阳性细胞数,观察分析SMS在白血病患者骨髓单个核细胞中CD34抗原表达的分布和含量.结果在相差倒置显微镜下,加入SMS组与对照组比较,细胞碎片增多,基质细胞减少;CD34抗原主要表达于细胞膜和核膜;SMS组阳性细胞数显著少于对照组(P＜0.01);图像分析光密度值(OD值)SMS组明显低于对照组(P＜0.01).结论在体外SMS可减少造血干/祖细胞特异抗原(CD34)的表达.提示SMS对白血病细胞的生长具有负性调控作用.     

组胺H2受体激动剂4—甲基组胺对阿糖胞苷抑制HL—60细胞增殖的负？…

在用４－甲基组胺（４－ＭＨ）激动ＨＬ－６０细胞上的组胺Ｈ２受体后,观察抗白血病药物阿糖胞苷（Ａｒａ－Ｃ）对ＨＬ－６０白血病细胞增殖抑制作用的影响。以＾３Ｈ－ＴｄＲ掺入和集落培养为增殖指标,用ＮＢＴ还原试验检测细胞的分化,用ＨＰＣＬ和荧光分光光度法分别检测细胞内ｃＡＭＰ和Ｃａ＾２＋离子的变化。结果显示,ＨＬ－６０细胞上的组胺Ｈ２受体经４－ＭＨ激动后,Ａｒａ－Ｃ对其增殖的抑制作用明显减弱。