益肾生血片治疗再生障碍性贫血的实验研究

目的：探讨益明生血片治疗再生障碍性贫血（再障）的作用机理。方法：观察了该药对小鼠造血功能，免疫功能，抗感染力和抵抗力的影响。结果：益肾生血片对马利兰引起的造血损伤小鼠多能造血干细胞（ＣＦＵ－Ｓ）、粒素祖细胞（ＣＦＵ－Ｃ）和红系祖细胞（ＣＦＵ－Ｅ）的微量的恢复有明显促进作用；对正常小鼠腹腔巨噬细胞吞噬功能有明显促进作用；可明显降低接种金黄色葡萄球菌小鼠的死亡率；在常压缺氧时，可使小鼠耐缺氧能力明显提     

Hematopoietic effect of fractions from the enzyme-digested colla corii asini on mice with 5-fluorouracil induced anemia

Effects of fractions A and B from enzyme-digested traditional Chinese medicine colla corii asini on mice with 5-fluorouracil-induced anemia were investigated. The purpose of this study was to further understand the hematopoietic activities and mechanisms of colla corii asini. The fractions A and B were administered to anemic mice for 12 days. After confirming the anti-anemic effect of fractions A and B, we examined the effects of fractions A and B on immature granulocyte and erythroid cell activity and plasma cytokine level. Fraction A administration at 2 g/kg and 1 g/kg and fraction B administration at 1.6 g/kg and 0.8 g/kg activated granulocyte and erythrocyte progenitor cells in bone marrow and erythrocyte progenitors in spleen, led to the recovery of white blood cell and red blood cell counts, and increased the percentage of peripheral reticulocytes in red cells. The GM-CSF and EPO production determined by examining GM-CSF mRNA and EPO mRNA in the kidney and liver of the anemic mice were also enhanced. This treatment significantly increased serum GM-CSF and EPO level and lowered serum transforming growth factor (TGF-beta) level. These results suggested that fractions A and B promoted hematopoiesis by activating immature granulocyte and erythroid cells, partly by stimulating GM-CSF and EPO secretion and suppressing TGF-beta release. Identification of a specific peptide or protein is still required for the development of a novel medicine for anemia caused by malignancy or chemotherapy.     

补肾益髓生血法AA大鼠含药血清对大鼠造血干细胞红系分化JAK2/STAT5信号通路的影响

目的:探讨补肾益髓生血法再生障碍性贫血(Aplastic Anemia,AA,简称再障)大鼠含药血清对大鼠骨髓造血干细胞红系分化JAK2/STAT5信号通路的影响。方法:在前期整体动物实验的基础上,体外培养正常大鼠造血干细胞,诱导其定向红系分化,分为空白对照组、正常对照组、模型组、司坦唑醇组(康力龙组)、益髓生血组、温肾生血组和滋肾生血组,在培养体系中加入含药血清干预4d后,提取红系细胞总RNA及蛋白,分别采用RT-PCR和Western blot检测细胞JAK2、STAT5mRNA及蛋白表达情况。结果:与正常对照组相比,模型组细胞JAK2、STAT5mRNA及蛋白表达均明显降低(P0.01);与模型组相比,各治疗组JAK2、STAT5mRNA及蛋白表达明显升高(P0.01或P0.05);滋肾生血组JAK2、STAT5mRNA及蛋白表达高于益髓生血组和温肾生血组(P0.05)。结论:补肾益髓生血法能够增强JAK2、STAT5的表达,可能通过影响JAK2/STAT5信号通路来促进红系细胞的分化成熟,其中滋肾生血法优于益髓生血法和温肾生血法。     

黄芪注射液对骨髓抑制性贫血小鼠造血调控的实验研究

目的观察黄芪注射液(Astragalus membranaceus injection,AMI)对放、化疗因素所致骨髓抑制性贫血小鼠造血的影响,并探讨其可能的机制.方法:采用Co60照射、注射环磷酰胺、氯霉素复合造模建立贫血小鼠模型,并将动物随机分为三组:AMI给药组Ⅰ、Ⅱ和对照组,分别腹腔注射AMI[500 mg/kg、1000 mg/kg]或等量生理盐水,第7日全自动血球仪检测外周血和骨髓有核细胞,并应用免疫组化和造血祖细胞体外培养技术,检测骨髓有核细胞BcL-XL和BcL-2蛋白的表达及进行三系造血祖细胞集落计数.结果:AMI给药组Ⅰ的RBC、HB、PLT(P＜0.05)和BM(P＜0.01)明显高于贫血对照组;同时给药组Ⅰ的CFU-E、BFU-E、CFU-Meg和骨髓有核细胞抗凋亡蛋白BcL-XL的表达也明显高于贫血对照组(P＜0.05或P＜0.01),给药组Ⅱ的CFU-E和骨髓有核细胞抗凋亡蛋白BcL-XL的表达也高于贫血对照组(P＜0.05).结论:黄芪注射液可以通过上调抗凋亡蛋白BcL-XL表达减轻骨髓有核细胞的凋亡,并促进红系、巨核造血.     

Radioprotective effects of dammarane sapogenins against 60Co‐induced myelosuppression in mice

Radiotherapy frequently induces failure of hematopoietic system and leads to myelosuppression. The objective of this study was to investigate the protective effect of dammarane sapogenins (DS), the hydrolysed product of the constituent ginsenosides of Panax ginseng, which are produced by gut metabolism, on radiation‐induced hematopoietic injury. Mice were exposed to 3.5 Gy ⁶⁰Co γ‐rays of total body radiation at a dose rate of 1.60 Gy per minute and treated with DS or granulocyte colony‐stimulating factor immediately after radiation. The general condition of the mice, the peripheral blood cell counts, multiple colony forming unit (CFU) assays of hematopoietic progenitor cells, hematopoietic stem cell counts, bone marrow histology, and spleen colony forming unit counts were then investigated. Our results indicated that administration with DS could ameliorate ⁶⁰Co‐irradiation induced damage and significantly increase the number of peripheral blood cells (white blood cells and platelets), 5 types of hematopoietic progenitor cells CFU (CFU‐GM, CFU‐E, BFU‐E, CFU‐Meg, and CFU‐GEMM), hematopoietic stem cell (Lin^?c‐kit⁺Scal‐1⁺) numbers, and CFUs in the spleen, as well as improved bone marrow histopathology. All together, these results confirmed the enhancement of DS on hematopoiesis.     

Experimental study of the bone marrow protective effect of a traditional Chinese compound preparation

This study investigated the effect of Wei Gan Li on bone marrow haemopoiesis in myelosuppressed anaemic mice induced by ⁶⁰Coγ, cyclophosphamide and chloramphenicol. After treatment with Wei Gan Li, colony forming units of granulocyte macrophages, erythroid cells, burst forming unit‐erythroid cells, megakaryocytes and the proliferation of bone marrow stromal cells were measured by in vitro cell culture techniques. Bone marrow haemopoietic stem cells were measured by flow cytometry. The peripheral blood cell count was found to have significantly recovered after Wei Gan Li administration and the proliferation of bone marrow haemopoietic stem/progenitor cells and BMSC were also significantly increased. Wei Gan Li was therefore found to promote the recovery of bone marrow haemopoietic function in myelosuppressed anaemic mice. Copyright © 2009 John Wiley & Sons, Ltd.     

人参总皂甙对正常人和再生障碍性贫血患者造血祖细胞的刺激增殖作用

为了解人参对造血祖细胞的作用,应用红系祖细胞(BFU-E、CFU-E)和粒单系祖细胞(CFU-GM)体外培养技术,观察人参总皂甙(TSPG)对正常人和29例再生障碍性贫血(再障)患者的祖细胞刺激增殖作用,并与甲基睾丸素(甲睾)相比较。TSPG在20μg/ml浓度时,可使正常BFU-E、CFU-E和CFU-GM产率分别提高37.8±2.9%、31.4±2.9%和33.4±4.0%。去除红细胞生成素,TSPG亦具有直接刺激红系祖细胞增殖作用,而甲睾对CFM-GM无作用。29例再障中雄激素反应型14例(48.1%),均同时呈现对人参的敏感性(集落数增加30%以上)。而免疫介导型及干细胞缺少型中人参的作用均不能表达。提示TSPG的作用机制可能是通过增强红细胞生成素等造血生长因子的活性间接地促进骨髓造血。     

三七总皂甙对免疫介导性再生障碍性贫血小鼠造血祖细胞的增殖作用

目的：探讨三七总皂甙(PNS)对免疫介导性再生障碍性贫血(再障)小鼠模型血细胞生成的作用。方法：采用免疫介导法进行再障造模,Balb／c小鼠经^60Coγ射线亚致死量照射后,自尾静脉输入DBA／2小鼠的淋巴细胞,随机分成4组,即模型组、PNS治疗高、中和低剂量组(每只3．2、1．6、0．8mg／d),并设正常对照组。腹腔注射给药,模型组和正常对照组注射生理盐水。治疗12天后,做血白细胞计数、骨髓病理检查和粒系、红系造血祖细胞(CFU-GM、CFU-E)集落培养。结果：(1)PNS升高白细胞数,与模型组比较,PNS高、中和低剂量组的白细胞数提高率分别为(34．3±2．9)％,(29．2±1．7)％和(14．5±1．6)％(P<0．01或P<0．05)。(2)改善骨髓抑制,骨髓病理检查模型组出现大片空白区,被大量的脂肪组织代替,而PNS治疗组造血组织结构较完整,造血细胞量丰富。(3)促进造血祖细胞增殖,高、中和低剂量PNS对CFU—GM集落提高率分别为(64．4±2．8)％、(67．3±2．4)％和(21．9±1．8)％(均P<0．01);CFU—E集落的提高率分别为(31．9±3．6)％、(20．7±2．4)％和(12．8±2．6)％(P<0．01或P<0．05)。结论：免疫介导性再障小鼠在骨髓抑制、造血功能低下时,PNS通过促进骨髓粒系、红系造血祖细胞的增殖,改善骨髓造血组织增生,从而促进血细胞的生成。     

慢性再生障碍性贫血的中医药研究

慢性再生障碍性贫血(CAA)是由多种病因引起的骨髓造血干细胞和骨髓微环境严重受损,以全血细胞减少为主要表现的一组综合征。中医认为其基本病机在于肾虚,肾不藏精、生髓,髓不化血,强调"以肾为本、从肾论治",临床上治疗慢性再生障碍性贫血疗效满意,文章就再生障碍性贫血从肾论治的中医理论基础、病因病机、辨证论治等方面进行综述。     

人参多糖和当归多糖诱导人内皮细胞表达造血生长因子的实验研究

目的：研究和论证人参和当归促进血细胞生成及“补气生血”的现代生物学机理。方法：采用造血生长因子生物活性检测与免疫细胞化学等技术，研究经人参多糖（GPS）和当归多糖（APS）诱导的人脐静脉内皮细胞株（Ecv304)上清液（HEcCM)中造血生长因子的活性及粒单系集落刺激因子（GM－CSF）、IL－3和IL－6在内皮细胞的表达。结果：GPS和APS体外诱导制备的HFcCM能显著促进粒单系造血祖细胞（CFU－GM）和多向性造血祖细胞（CFU－Mix)的增殖；同时，GPS和APS能不同程度地促进内皮细胞GM－CSF、IL－3和IL－6蛋白表达。结论：GPS和APS能上调造血微环境中的内皮细胞分泌造血生长因子来调节血细胞的生成。     

补髓生血胶囊治疗慢性再生障碍性贫血的临床观察

探讨补髓生血胶囊治疗慢性再生障碍性贫血的理论依据。采用造血细胞培养和流式细胞仪技术，对口服补髓生血胶囊的３５例患者治疗４个月前后的骨髓单个核细胞进行对比观察。１；本品具有恢复造血干／祖细胞膜ＩＬ－３，ＩＬ－６，ＩＬ－１１受体的作用；２．补髓生血胶囊的疗效优于西药；３．治疗组中肾阳虚与肾阴虚型治疗前的造血干／祖细胞在ＩＬ－３，ＩＬ－６，ＩＬ－１１刺激下，其细胞集落产生率和ＣＤ＾＋３４细胞率有显著性差     

复方活血汤对免疫诱导再生障碍性贫血小鼠骨髓微环境的作用研究

目的 :探讨临床上治疗再生障碍性贫血 (再障 )加用活血化瘀药物提高疗效的机制。方法 :建立免疫介导的再障小鼠模型 ,胃饲 1 0 0 %复方活血汤注射液每次 0 2ml,每天 2次 ,第 1 0天观察骨髓组织学、体外成纤维细胞集落形成单位 (CFU -F)、培养基质细胞层的粘附能力 ,骨髓氧分压 (PO2 )。结果 :复方活血汤组小鼠白细胞计数 ,骨髓有核细胞计数 ,骨髓造血组织容量、CFU -F计数 ,均较再障组有明显升高 (P <0 0 1 ) ;且基质细胞粘附功能、骨髓PO2 已恢复至正常。结论 :复方活血汤可促进再障小鼠骨髓微环境的修复及供氧 ,从而促进骨髓造血。     

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??OBJECTIVE To study the blood enriching effect and mechanism of Jiuweiejiao ointment on mouse model of blood deficiency syndrome.METHODS Blood deficiency syndrome model was established with cyclophosphamide(CTX) and acetylphenylhydrazine(APH). Total of 60 ICR mice were divided into normal group, model group, Siwu granule group, Jiuweiejiao ointment high-dose, medium-dose and low-dose group (15.6, 7.8, 3.9 g??kg^-1). Each group mouse were treated with the respective drug through intragastrical administration once a day for consecutive 14 d. The levels of red blood cells (RBC), leukocytes (WBC), hemoglobin (HGB) level in blood of mice were detected by blood analyzer; the histopathological changes of bone marrow were determined by HE staining; the levels of erythropoietin (EPO), interleukin-3 (IL-3), tumor necrosis factor-?? (TNF-??), granulocyte colony stimulating factor (G-CSF) and granulocyte-macrophage colony stimulating factor (GM-CSF) in serum were measured by enzyme linked immunosorbent assay (ELISA) method; the expression level of EPO protein in kidney tissue, G-CSF and GM-CSF protein in bone marrow tissue were detected by Western-blot method.RESULTS Compared with model group, Jiuweiejiao ointment could significantly increase the levels of RBC, WBC and HGB in blood of mice, markedly improve bone marrow hyperplasia, reduce the non-hematopoietic tissue and adipocytes, obviously repaire the hematopoietic tissue structure; significantly decrease the level of TNF-?? and increase the level of EPO, IL-3, G-CSF and GM-CSF in serum. In addition, Jiuweiejiao ointment could also obviously increase the expression of EPO protein in kidney tissue and G-CSF, GM-CSF protein in bone marrow tissue compared with that in model group.CONCLUSION Jiuweiejiao ointment has obvious blood enriching effects, which may be related with promoting the secretion of hematopoietic growth factors and reducing the inhibition of hematopoietic function by negative hematopoietic factor.     

扶正祛邪丹治疗骨髓增生异常综合征的临床及实验研究

目的：研究扶正祛雅丹对骨髓增生异常综合征（ＭＤＳ）的疗效与作用机理。方法：用扶正祛邪丹为主治疗ＭＤＳ患者３３例，观察血象，骨髓象，骨髓病理切片及免疫学的变化，用干细胞培养法观察扶正祛邪丹对ＢＡＬＢ／Ｃ小鼠骨髓造血干细胞增殖的影响。结果：临床患者基本缓解率２４．２４％，总有效率７２．７２５，实验研究发现扶正祛邪丹对骨髓红系造血干细胞有促进作用。结论：扶正祛邪丹对ＭＤＳ的难治性贫血疗效较好，可能是通过     

刺五加注射液对^60Co-γ放射损伤小鼠骨髓CFU—E、BFU—E生成的影响

目的观察刺五加注射液对^60Co-γ放射损伤小鼠骨髓红系集落形成单位（CFU—E）和红系爆式集落形成单位（BFU—E）生成的影响。方法小鼠接受6．0Gy^60Co-γ射线照射后立即分别腹腔注射刺五加注射液0．2和0．4ml／10g，均每132次，连续3d；照射后第4日取股骨骨髓制备骨髓有核细胞悬液，甲基纤维素半固体培养法测定CFU—E和BFU—E集落。结果刺五加注射液能明显提高有核细胞CFU—E和BFU—E集落数量，与模型组相比均具有显著差异。结论刺五加注射液能明显提高放射损伤小鼠骨髓有核细胞数量及红系祖细胞集落形成，从而保护受损小鼠骨髓造血功能。     

Comparative investigations on the protective effects of rhodioside, ciwujianoside-B and astragaloside IV on radiation injuries of the hematopoietic system in mice

The aim of this study was to investigate the protective effects of three glycosides (rhodioside, ciwujianoside‐B and astragaloside IV) on the hematopoietic system in the mice exposed to γ‐rays, and to examine the possible mechanisms involved. Mice were pretreated with the glycosides (40 mg/kg, i.g.) daily for 7 days prior to radiation. The survival of mice pretreated with three glycosides after total body irradiation (6.0 Gy) was examined. Peripheral blood leucocytes and endogenous spleen colony counts, colony‐forming unit‐granulocyte macrophage assay, analysis of DNA content and apoptosis rate determination were performed to evaluate the effects of the three glycosides on hematogenesis. The fragmentation of double‐stranded DNA in lymphocytes was detected by the comet assay. The changes in cell cycle were analysed by flow cytometry. Furthermore, the expression levels of Bcl‐2, Bax and nuclear factor‐kappa B (NF‐κB) were measured by western blot and the electrophoretic mobility shift assay. The results showed that pretreatment with all of the glycosides improved survival time and increased the number of leucocytes, spleen colonies and granulocyte‐macrophage colonies in mice exposed to 6.0 Gy γ‐radiation. Rhodioside showed more protective efficacy than both ciwujianoside‐B and astragaloside IV. All three glycosides significantly increased the proliferation abilities of bone marrow cells, and decreased the ratio of cells in G₀/G₁ phase. Further analysis showed that these three glycosides were able to decrease DNA damage and the increment in the Bax/Bcl‐2 ratio induced by radiation. In summary, the three glycosides showed radioprotective effects on the hematopoietic system in mice, which was associated with changes in the cell cycle, a reduction in DNA damage, and down‐regulation of the ratio of Bax/Bcl‐2 in bone marrow cells exposed to radiation. Copyright © 2010 John Wiley & Sons, Ltd.     

补肾泻肝方对再生障碍性贫血造血与免疫失调治疗的实验研究

目的:证实补肾泻肝方对再生障碍性贫血造血与免疫失调的治疗作用。方法:利用免疫介导再生障碍性贫血动物模型,观察补肾泻肝方对再生障碍性贫血造血与免疫失调的治疗作用。结果:用药组抑制细胞活性,CD_3~ 细胞和 CD_8~ 细胞较对照组明显减低(P<0.05):用药组小鼠骨髓有核细胞数、Hb、WBC BPC 在第12天均高于对照组(P<0.05);病理组织学显示用药组至第12天骨髓见散在增生灶,第60天基本恢复正常。结论:提示补肾泻肝方具有明确的解除 CD_8~ 细胞对造血的抑制,同时促进造血干细胞生长的作用。     

双黄升白颗粒剂对白细胞减少症模型小鼠造血细胞作用的研究

目的:探讨双黄升白颗粒剂升高白细胞作用及其作用机制。方法:采用Se-137放射源照射ICR小鼠,造成白细胞减少症动物模型,对模型动物给予双黄升白颗粒剂大、中、小剂量(分别为9.2,4.6,2.3 g.kg^-1.d^-1)灌胃,并设正常对照组,模型对照组,阳性对照利可君组。对小鼠外周血象,骨髓有核细胞(BMNC),脾结节(CFU-S)计数,胸腺指数(TI),脾脏指数(SI),骨髓细胞增殖,体外培养小鼠粒单系集落形成单位(CFU-GM),骨髓CD34+细胞比例,骨髓造血微环境等进行检测。结果:双黄升白颗粒可明显升高模型小鼠的白细胞(WBC);双黄升白颗粒可升高模型小鼠的BMNC计数并促进其骨髓细胞增殖;双黄升白颗粒升高模型小鼠CFU-S计数,提高CFU-GM产率,提高骨髓CD34+细胞比例,对造血干/祖细胞增殖具促进作用;双黄升白颗粒具有保护模型小鼠骨髓造血微环境作用;双黄升白颗粒升高模型动物SI,具有一定的免疫增强作用。结论:双黄升白颗粒剂具有明显升高白细胞作用,其作用机制与促进造血细胞增殖及保护骨髓造血微环境有关。     

中药配合脐血输注治疗再生障碍性贫血的临床观察

４８例再生障碍性贫血分为两组。治疗组３０例采用中药配合脐血输注治疗，对照组１８例采用单纯脐血输注治疗。治疗组与对照组总有效率分别为９３．３％，与７２．２％。两组比较有显著性差异。两组再障患者治疗前后骨髓粒－单系祖细胞体外培养结果表明，两组患者治疗３个月骨髓ＣＦＵ－ＧＭ增高水平接近，治疗后６个月治疗组骨髓ＣＦＵ－ＧＭ持续稳定上升，而对照组治疗后６个月与治疗后３个月比较有所下降，提示治疗组骨髓造血的恢