当归多糖对人粒单系造血祖细胞增殖分化的调控机理研究

目的：研究当归多糖（APS）对人粒单系血细胞发生的影响及其调控的机理。方法：采用造血祖细胞体外培养,造血生长因子生物活性检测,核酸分子原位杂交和免疫细胞化学等实验血液学技术,研究ASP对人粒单系造血祖细胞（CFU－GM）增殖分化的影响。结果：APS在体外能显著刺激CFU－GM的增殖;经APS诱导制备的人胸腺细胞,腺细胞、骨髓基质细胞条件培养液能明显促进CFU－GM增殖,经APS体外刺激后骨髓基质细胞、脾细胞和胸腺细胞的GM－CSF蛋白和mRNA表达水平显著提高。结论：APS可能通过直接或间接途径促进淋巴细胞和造血微环境中的基质细胞合成和分泌GM－CSF或GM－CSF样物质,进而促进粒单系血细胞的发生。     

人参多糖对粒单系造血祖细胞增殖分化的影响

本文采用造血祖细胞体外培养、造血生长因子生物活性检测、免疫细胞化学等实验血液学技术检测人参多糖(GPS)体外作用对人粒单系造血祖细胞(CFU—GM)增殖分化的影响及GPS刺激制备的胸腺细胞培养上清液(HTCM)、脾细胞培养上清液(HSCM)、骨髓基质细胞条件培养液(HBMSCM)中GM-CSF的生物活性。结果发现：①在有外源性粒单系集落刺激因子(GM—CSF)存在的情况下,GPS能显著促进CFU—GM的增殖与分化。②经GPS体外诱导制备的胸腺细胞、脾细胞、骨髓基质细胞的条件培养液能明显提高人CFU—GM的产率。③经GPS体外刺激后,胸腺细胞、脾细胞、骨髓基质细胞GM-CSF蛋白表达水平较对照组明显提高。结果表明GPS可能通过直接和/或间接途径促进胸腺细胞、脾细胞和造血诱导微环境中的基质细胞合成和分泌GM—CSF或GM-CSF样活性,进而促进CFU-GM的增殖和分化。     

人参总皂甙诱导淋巴细胞表达GM-CSF的实验研究

目的：研究人参总皂甙(TSPG)对人淋巴细胞表达GM-CSF的影响，及其与人粒细胞．巨噬细胞系血细胞发生调控的关系。方法：采用造血祖细胞、胸腺细胞和脾细胞体外培养、造血生长因子生物学活性检测、免疫细胞化学和核酸分子原位杂交等技术。结果：TSPG能显著刺激粒细胞，巨噬细胞系造血祖细胞(CFU-GM)增殖；经TSPG诱导制备的胸腺细胞和脾细胞条件培养液富含GM-CSF样活性；TSPG能显著促进胸腺细胞和脾细胞的GM-CSF蛋白和，mRNA表达。结论TSPG可能通过直接和／或间接途径促进淋巴细胞合成和分泌GM-CSF，进而促进CFU-GM的增殖分化。     

当归多糖诱导L—细胞产生造血生长因子的实验研究

用造血祖细胞体外培养，造血生长因子检测和流式细胞术等方法。研究当归多糖诱导Ｌ－细胞（成纤细胞株）产生造血和在子及其对知细胞姓的调节作用，结果表明，Ｌ－细胞条件培养液对造血祖细胞的体外增殖呈双向调节作用；１０％ＬｃＣＭ能促进骨髓造血细胞进入增殖活跃的Ｓ＋Ｍ／Ｇ２期；经ＡＰ诱导制备的ＬｃＣＭ的有或无外源性造血生长因子存在时均对造血祖细胞的克隆增殖显示较高刺激活性；ＡＰ或ＩＬ－１与造血生长因子间似无协同     

当归多糖对人髓系多向造血祖细胞增殖分化的影响及其机理研究

目的 :研究“补血活血”要药当归的主要有效成分 当归多糖 (APS)对人早期血细胞发生的影响及其调控机理。方法 :采用造血祖细胞体外培养、造血生长因子生物学活性检测、免疫细胞化学、核酸探针原位杂交等实验技术。结果 :APS在体外可促进髓系多向造血祖细胞CFU GEMM增殖分化 ;APS诱导制备的骨髓基质细胞、内皮细胞、单核细胞培养上清液富含造血生长因子活性 ;APS可促进骨髓基质细胞、内皮细胞、单核细胞表达GM CSF、IL 3蛋白 ,并对骨髓基质细胞表达GM CSF、IL 3mRNA有上调作用。结论 :APS可能通过促进造血微环境中的基质细胞表达和分泌GM CSF、IL 3等造血生长因子 ,促进人早期造血细胞发生 ,这可能是当归“补血活血”的分子生物学机理之一     

人参总皂甙对骨髓巨噬细胞的影响及与造血调控的关系

目的：研究人参总皂甙(TSPG)对骨髓巨噬细胞(BMMφ)生物学活性的影响及其与造血调控关系。方法：采用骨髓造血祖细胞体外培养，造血生长因子生物活性检测，免疫细胞化学，核酸探针原位杂交等技术。结果：经TSPG诱导制备的BMMφ的培养上清可提高髓系造血祖细胞的集落产率；经TSPG诱导后，BMMφ表达EPO，IL-3，IL-6，GM-CSF的蛋白水平有不同程度提高；EPO mRNA，GM-CSF mRNA的表达水平和强度明显提高。结论：TBPG可以刺激造血诱导微环境中的巨噬细胞，从基因水平和蛋白水平2级层次上促进造血调控因子的合成和分泌，进而促进CFU-Mix，CFU-E，CFU-GM的增殖分化。     

骨髓增生异常综合征的造血抑制活性

采用体外单层琼脂培养技术，观察了10例骨髓增生异常综合征(MDS)患者骨髓和外周血单个核细胞条件培养液和血清对正常骨髓粒单系祖细胞集落形成影响．并与16例急性髓细胞性白血病(AML)患者进行比较．结果表明，与AML-样，MDS患者骨髓、外周血单个核细胞条件培养液和血清可抑制粒单系祖细胞集落形成，存在白细胞和关抑制活性和血清抑制活性，推测这种抑制活性可能与患者粒细胞减少有关。     

当归多糖诱导L－细胞产生造血生长因子的实验研究

用造血祖细胞体外培养、造血生长因子检测和流式细胞术等方法,研究当归多糖（ＡＰ）诱导Ｌ－细胞（成纤维细胞株）产生造血生长因子及其对血细胞发生的调节作用。结果表明,Ｌ－细胞条件培养液（ＬｃＣＭ）对造血祖细胞（ＣＦＵ－ＧＭ、ＢＦＵ－Ｅ、ＣＦＵ－Ｅ）的体外增殖呈双向调节作用;１０％ＬｃＣＭ能促进骨髓造血细胞进入增殖活跃的Ｓ＋Ｍ／Ｇ２期;经ＡＰ诱导制备的ＬｃＣＭ在有或无外源性造血生长因子（如Ｅｐｏ,ＢＰＡ,ＧＭ－ＣＳＦ）存在时均对造血祖细胞的克隆增殖显示较高刺激活性;ＡＰ或ＩＬ－１与造血生长因子间似无协同作用。本研究提示ＡＰ可能通过诱导造血微环境的成纤维细胞分泌某些造血生长因子,从而促进造血祖细胞增殖分化,这或许是当归＂补血＂的生物学机理之一。     

当归补血汤载药血清干预后骨髓基质细胞中粒细胞集落刺激因子和白细胞介素3的分泌

背景：实验表明,当归补血汤的有效成分多糖对造血干细胞和造血祖细胞的增殖分化存在显著的促进作用,对造血微环境具有重要的调控作用。 目的：观察当归补血汤载药血清干预后小鼠骨髓基质细胞中粒细胞集落刺激因子和白细胞介素3的分泌状况。 方法：分离骨髓基质细胞,于96孔板培养,在相差显微镜下观察细胞形态和生长状况;将细胞分4组,加含有不同剂量的当归补血汤载药血清进行干预,采用MTT法检测各组细胞的增殖活性,ELISA法检测粒细胞集落刺激因子和白细胞介素3的表达情况。 结果与结论：当归补血汤载药血清可明显促进骨髓基质细胞的增殖,促进骨髓基质细胞分泌粒细胞集落刺激因子和白细胞介素3,呈剂量依赖性,含等效剂量载药血清组促增殖作用和促分泌作用优于其他各组,浓度过高可减弱细胞的增殖分泌效应。     

人外周血LAK细胞对粒单系、红系祖细胞体外增殖的影响

43例人外周血LAK细胞对急性白血病细胞有较强的杀伤活性。LAK细胞对正常人骨髓粒单系、红系祖细胞无抑制作用,适宜浓度(20％)的LAK细胞培养上清可促进正常人骨髓粒单系、红系祖细胞的体外增殖。提示LAK细胞输注对白血病病人的正常造血细胞无损伤。     

Polysaccharides from the root of Angelica sinensis protect bone marrow and gastrointestinal tissues against the cytotoxicity of cyclophosphamide in mice

Cyclophosphamide (CY) is a cytostatic agent that produces systemic toxicity especially on cells with high proliferative capacity, while polysaccharides from Angelica sinensis (AP) have been shown to increase the turnover of gastrointestinal mucosal and hemopoietic stem cells. It is not known whether AP has an effect on CY-induced cytotoxicity on bone marrow and gastrointestinal tract. In this study, we assessed the protective actions of AP on CY-induced leukopenia and proliferative arrest in the gastroduodenal mucosa in mice. Subcutaneous injection of CY (200 mg/kg) provoked dramatic decrease in white blood cell (WBC) count and number of blood vessels and proliferating cells in both the gastric and duodenal mucosae. Subcutaneous injection of AP significantly promoted the recovery from leukopenia and increased number of blood vessels and proliferating cells in both the gastric and duodenal tissues. Western blotting revealed that CY significantly down-regulated the protein expression of vascular endothelial growth factor (VEGF), c-Myc and ornithine decarboxylase (ODC) in gastric mucosae but had no effect on epidermal growth factor (EGF) expression. AP also reversed the dampening effect of CY on VEGF expression in the gastric mucosa. These data suggest that AP is a cytoprotective agent which can protect against the cytotoxicity of CY on hematopoietic and gastrointestinal tissues when the polysaccharide is co-administered with CY in cancer patients during treatment regimen.     

当归多糖对小鼠腹腔巨噬细胞释放细胞效应分子的诱导作用

目的 :观察当归多糖对小鼠腹腔巨噬细胞产生细胞毒效应分子的影响。方法 :从BALB/c小鼠的腹腔分离巨噬细胞 ,并进行原代细胞培养。采用MTT比色法及紫外分光光度法 ,检测当归多糖对腹腔巨噬细胞释放一氧化氮 (NO)、肿瘤坏死因子α(TNF α)、活性氧 (ROS)以及诱导型一氧化氮合酶 (iNOS)和溶菌酶 (LSZ)活性的影响。结果 :当归多糖可激活巨噬细胞释放NO、TNF α和ROS等效应分子 ,并显著提高LSZ的活性。当归多糖可能通过提高巨噬细胞iNOS的活性而增加NO的释放量 ,但其与脂多糖 (LPS)无协同促进NO释放的作用。当归多糖体外无直接杀伤肿瘤细胞的作用 ,但其与巨噬细胞共孵育的培养上清具有杀伤L92 9细胞的作用。结论 :当归多糖可促进巨噬细胞释放NO、TNF α及ROS等细胞效应分子 ,并可通过作用于巨噬细胞而促进TNF α的分泌 ,发挥间接的抗肿瘤免疫作用     

Effects of postirradiation carboxymethylglucan administration in mice

The hemopoiesis-enhancing ability of a soluble glucan derivative, i.e. carboxymethylglucan (CMG), was investigated in gamma-irradiated mice. Attention was focused on the usefulness of its single or repeated postirradiation administration. CMG was administered i.p. at (a) single dose of 6 mg 2 h postirradiation, (b) four 6 mg doses in the first 4 days postirradiation, (c) four 1.5 mg doses at the same time intervals. Indices of granulopoiesis and inflammatory side effects (liver weight increase and hepatic granulomas) were investigated in mice irradiated with a sublethal dose of 7 Gy. All three CMG-treated groups of mice were found to exhibit enhanced hemopoietic recovery in comparison with the controls. Although the mice repeatedly given the 6 mg CMG doses showed the most rapid recoveries of all the evaluated parameters of granulopoiesis, the most pronounced hepatic side effects were found in these mice, too. When survival of mice was recorded in lethally (9 Gy) irradiated animals, the best protective response were obtained following the repeated administration of the 1.5 mg CMG dose, the survival by day 30 in this group being significantly higher not only in comparison with the controls but also with the mice repeatedly given the 6 mg dose of CMG. The results suggest that the postirradiation CMG administration can be useful for enhancing radiation suppressed hemopoiesis. However, repeated larger CMG doses may produce side effects which compromise the overall survival of irradiated mice.     

当归注射液对辐射损伤后肝组织中超氧化物歧化酶活性的影响

目的　探讨当归注射液对辐射损伤后肝组织中超氧化物歧化酶活性的影响。方法　应用当归注射液对60 CO γ射线辐射损伤后的小鼠肝组织给予治疗 ,连续给药 30d ,应用放射免疫测定法检测治疗前后肝组织中超氧化物歧化酶活性。结果　实验用药组肝组织中超氧化物歧化酶活性高于实验对照组 (P <0 0 5 )。结论　当归注射液对辐射损伤后的小鼠肝组织和功能具有重要的保护作用     

Mechanisms of hemopoiesis in athymic mice

Cell elements of peripheral blood and bone marrow and splenic hemopoiesis were studied in athymic BALB/c mice (nude or thymectomized animals). In athymic animals lympho- and erythropoiesis were suppressed, while granulocyto- and monocytopoiesis were activated. These changes were more pronounced in nude mice.     

Humoral control of monocytopoiesis by an activator and an inhibitor

Summary During an inflammatory reaction, a factor increasing monocytopoiesis (FIM) was found in the serum in the early phase of an acute inflammation and for a longer period in a prolonged inflammation. FIM, which is a protein with a molecular weight of about 20,000 Daltons, exerts its action by decreasing the cell-cycle time of the promonocytes and concomitantly increasing the number of these cells, in all likelihood due to an effect on the monoblasts. FIM is released from the cells at the site of the inflammatory lesion, most probably from the macrophages.In addition, a monocytopoiesis inhibiting factor (MPI) was found. This factor, which has a molecular weight of more than 50,000 Daltons, has an effect on the proliferation of monoblasts.On the basis of these results, a system for the control of monocytopoiesis is postulated.     

Development of the liver in the chicken embryo. II. Erythropoietic and granulopoietic cells

Hepatic hemopoiesis is apparent in the chicken embryo on day 7 of incubation (Hamburger and Hamilton Stage 30), and a peak in hemopoietic activity occurs on day 14 (Stage 40). During this period, the differentiation of hemopoietic cells was examined by light microscopy and by transmission and scanning electron microscopy. Glycol methacrylate sections were used in lieu of smears to study hemopoietic cells, thus minimizing the problems of cell shrinkage and rupture. The sections were superior to smears for close examination of nuclear and cytoplasmic morphologies and for precise localization of hemopoietic cells to intravascular and extravascular sites. The avian liver is involved directly with erythropoiesis and granulopoiesis only. Erythropoietic cells, occurring in intravascular and extravascular locations, appear throughout the time frame examined. Blood islands with granulopoietic cells were not observed until days 8–9 (Stage 35). Granulopoiesis in the liver produces only eosinophilic leukocytes. Individual granulopoietic cells appear first in the connective tissue sheaths of hepatic vessels, and these cells subsequently congregate into blood islands. Endothelial cells of the sinusoidal linings, through asymmetric divisions, frequently release daughter cells into the circulation, and Kupffer cells are actively engaged in phagocytosis of erythrocytes. From a comparative standpoint the elements deemed critical to hemopoiesis in the mammalian liver–prehepatocyte population, hepatic vasculature, and compartments for stem cell differentiation–may not hold the same importance in the bird, owing to an inordinate reliance on intravascular hemopoiesis in this vertebrate class. © 1993 Wiley-Liss, Inc.     

Glucan: mechanisms involved in its "radioprotective" effect

It has generally been accepted that most biologically derived agents that are radioprotective in the hemopoietic-syndrome dose range (eg, endotoxin, Bacillus Calmette Guerin, Corynebacterium parvum, etc) exert their beneficial properties by enhancing hemopoietic recovery and hence, by regenerating the host's ability to resist life-threatening opportunistic infections. However, using glucan as a hemopoietic stimulant/radioprotectant, we have demonstrated that host resistance to opportunistic infection is enhanced in these mice even prior to the detection of significant hemopoietic regeneration. This early enhanced resistance to microbial invasion in glucan-treated irradiated mice could be correlated with enhanced and/or prolonged macrophage (but not granulocyte) function. These results suggest that early after irradiation glucan may mediate its radioprotection by enhancing resistance to microbial invasion via mechanisms not necessarily predicated on hemopoietic recovery. In addition, preliminary evidence suggests that glucan can also function as an effective free-radical scavenger. Because macrophages have been shown to selectively phagocytize and sequester glucan, the possibility that these specific cells may be protected by virtue of glucan's scavenging ability is also suggested.