粒细胞集落刺激因子动员骨髓干细胞治疗大鼠急性心肌梗塞

目的:探讨G-CSF动员的骨髓来源干细胞对急性心肌梗塞动物模型的治疗作用与对缺血濒死心肌的保护作用。方法:用异丙肾上腺素(ISO)复制急性心肌梗塞大鼠动物模型,于3h后用G-CSF动员骨髓干细胞释放和迁移至心肌梗塞部位,并分别于24h、48h和2周后杀死大鼠,取出心脏,检测心肌的病理变化情况。结果:用ISO后24h对照组可见散在心肌梗塞灶,坏死区周围有多量以中性粒细胞为主的炎症细胞浸润,而治疗组大鼠心肌坏死程度较对照组轻,浸润的细胞以单个核细胞为主;48h后对照组心肌梗塞灶进一步扩大,呈散在片状分布,而治疗组心肌梗塞灶扩大不明显,呈散在灶性分布,并可见成堆或散在分布的淋巴细胞样细胞;2周后,治疗组未见明显心肌梗塞后疤痕组织,可见新生的心肌细胞生长。结论:G-CSF对缺血濒死心肌有保护作用,用G-CSF动员骨髓来源的干细胞进行"自我移植",可用于急性心肌梗塞的治疗。     

粒细胞集落刺激因子对诱导成骨的作用

背景：间充质干细胞是一种具有自我复制能力和多向分化潜能的成体干细胞,在特定条件下能够发育成骨、软骨和其他类型的细胞,而粒细胞集落刺激因子可以促进间充质干细胞的再生。 目的：通过动物实验来观察粒细胞集落刺激因子对成骨的诱导作用,以期对临床骨缺损的患者提供一种更好的治疗方法。 方法：取新西兰白兔40只,随机等分为治疗组及模型组,均切除右侧约1 cm长度桡骨及周围骨膜,之后在缺损处植入脱钙骨基质。治疗组选用粒细胞集落刺激因子30 µg/(kg•d)皮下注射,于造模前3 d开始,连续应用7 d;模型组给予生理盐水皮下注射。 结果与结论：治疗组兔骨缺损区间充质细胞、成骨细胞数量和成骨质量上均优于模型组。提示粒细胞集落刺激因子的应用可以明显提高骨缺损区域的间充质干细胞数量,促进成骨,从而可以促进骨缺损的早期修复。 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程全文链接：     

Granulocyte colony-stimulating factor exacerbates the acute lung injury and pulmonary fibrosis induced by intratracheal administration of bleomycin in rats

We investigated the effects of granulocyte colony-stimulating factor (G-CSF) on lung injury induced by intratracheal administration of bleomycin (BLM, 2 mg/200 micro1) in rats. In experiment 1, G-CSF (10, 30 and 100 microg/kg/day, s.c.) was administered to rats treated with BLM or saline for 7 days starting immediately after BLM administration. In rats receiving G-CSF alone, a large number of neutrophils were noted in the pulmonary capillaries, although there were no lung lesions. In rats receiving BLM alone, diffuse alveolar damage was observed. The administration of G-CSF to BLM-treated rats increased the total lung lesion per unit of pulmonary parenchyma (total lung lesion %) along with increases in the peripheral neutrophil count and the number of neutrophils infiltrating in the pulmonary lesion in a dose-dependent fashion. In experiment 2, 100 microg/kg/day of G-CSF was administered to rats treated with BLM or saline for up to 28 days starting immediately after BLM administration. The administration of 100 microg/kg/day of G-CSF to BLM-treated rats showed no effects at 14 days but it increased the lung lesion % and the score of lung fibrosis along with the increase in the number of neutrophils infiltrating in the pulmonary lesion at 28 days. These findings suggest that G-CSF administration to BLM-treated rats influenced and exacerbated the BLM-induced acute lung injury, and also exacerbated pulmonary fibrosis in a dose-dependent fashion. The exacerbation of lung injury coincided with the marked increase in the peripheral neutrophil count and the number of neutrophils infiltrating in the pulmonary lesion.     

Granulocyte antibodies in Korean neonates with neutropenia

Neonatal alloimmune neutropenia (NAN) is a disease that can cause severe and prolonged neutropenia in neonates. However, no report is available on the incidence of granulocyte antibody in neonates, the target antigen of this antibody, and the estimated incidence of NAN in Korea. Among a total of 856 neonates admitted to a neonatal intensive care unit (NICU) over a five year period, a total of 105 neonates with neutropenia were enrolled in this study. Positive reactions were observed in the sera of six neonates (5.7%, 6/105) by mixed passive hemagglutination assay (MPHA). To confirm the presence of NAN, MPHA and granulocyte antigen typing (HNA-1a, -1b, -2a, -4a, and -5a) were performed on neonatal and maternal blood. To differentiate granulocyte antibody and HLA antibody, MPHA was also performed using HLA antibody adsorbed serum. We confirmed three cases (2.9%, 3/105) of NAN among neonates with neutropenia in which granulocyte antibody specificities (two anti-HNA-1b and one anti-HNA-1a) and fetomaternal granulocyte antigen mismatches were identified. In this study, the estimated incidence of NAN was 0.35% (3/856) among neonates admitted to NICUs in Korea.     

促红细胞生成素联合粒细胞集落刺激因子对大鼠缺氧心肌细胞的保护效应研究

目的 探讨促红细胞生成素(EPO)联合粒细胞集落刺激因子(G-CSF)对大鼠缺氧心肌细胞的保护机制.方法 采用胰酶消化大鼠心肌细胞并传代,用150 μmol/L浓度的CoCl2处理心肌细胞,模拟缺氧条件.采用流式细胞仪检测不同EPO+ G-CSF浓度下心肌细胞的凋亡和坏死水平.筛选出最适浓度后,将实验组分为对照组、缺氧组、缺氧+ EPO组、缺氧+G-CSF组和缺氧+EPO+ G-CSF组,Western blot法检测乏氧诱导因子(HIF)-1α、P53、PARP蛋白的表达变化.结果 心肌细胞在10 U/mLEPO+200 ng/mL G-CSF培养下,心肌细胞凋亡和坏死水平最低,故认为是最适浓度(F=14.73、11.95,P ＜0.05).和对照组相比,HIF-1α、P53和PARP蛋白在各实验组中表达水平均升高,HIF-1α以缺氧组升高最显著(F=22.65,P＜0.05),P53以缺氧+EPO+ G-CSF组升高最显著(F =25.18,P＜0.05).除对照组外,PARP蛋白在实验组中呈现逐渐降低趋势(F=17.48,P＜0.05).结论 EPO-G-CSF能降低缺氧的心肌细胞凋亡、坏死和DNA损伤,其机制可能与HIF-1α、P53、PARP蛋白表达相关.     

Inhibition of resistance to hemopoietic allo-grafts in granulocyte colony-stimulating factor transgenic mice

Transplanted allogeneic marrow cells often fail to engraft in a lethally irradiated host. This phenomenon, termed resistance to allogeneic marrow grafts or allo-resistance, is well documented, although its mechanism is not yet understood. Transplantation of major histocompatibility complex disparate allogeneic marrow cells into mice transgenic for granulocyte colony-stimulating factor (G-CSF) showed donor-derived spleen colonies (CFU-S) and resulted in stable allogeneic chimerism with excellent survival (100% up to 40 days and 89% up to 120 days). Under the same experimental conditions, all the littermate controls failed to show CFU-S and died shortly after marrow transplantation. Thus, resistance to allogeneic marrow cells appeared to be severely impaired in this transgenic mouse. The observation that neutralizing antibody against G-CSF restored allo-resistance in G-CSF transgenic mice and that CFU-S was inducible upon administration of recombinant G-CSF using a mini-osmotic pump in non-transgenic recipients, suggests that an elevated level of this cytokine is important for the inhibition of allo-resistance. Thus, G-CSF was found to play a role in allogeneic resistance to marrow grafts and the G-CSF-transgenic mice provide a useful model to study the inhibition of the resistance. The inhibition of allo-resistance may be useful in preparing allogeneic bone marrow chimeras in both experimental and clinical settings.     

下肢缺血及G-CSF促进小鼠外周血内皮祖细胞动员

目的：观察下肢缺血以及腹腔注射粒细胞集落刺激因子（G-CSF）对小鼠外周血内皮祖细胞（EPCs）的影响。 方法： 建立小鼠下肢缺血模型，双色荧光标记流式细胞技术检测缺血状态下外周血内皮祖细胞的变化，同时给予G-CSF后观察对EPCs的影响。 结果： 下肢缺血后，外周血中反映EPCs变化的CD34和VEGFR2双荧光阳性细胞比例轻度增加，在缺血基础上同时给予G-CSF，上述变化更明显。 结论： 下肢缺血刺激可以使外周血内皮祖细胞数量增加，G-CSF通过骨髓动员可增强这种效应。     

早产患者血清粒细胞集落刺激因子的检测及其意义

目的探讨粒细胞集落刺激因子(G-CSF)与自发性早产的关系及其对预测早产的可能意义。方法采用ELISA-双抗体夹心法分别对28-34周、34-37周不同孕龄组早产妇女及正常妊娠妇女的血清G-CSF进行检测。结果孕28-34周早产妇女血清G-CSF水平较正常对照组妇女升高(P<0.05)。结论高水平的血清G-CSF与妊娠<34周发生的早产有关,检测妊娠妇女血清G-CSF水平对预测妊娠34周之前发生的早产有重要意义。     

应用粒-巨噬细胞集落刺激因子治疗小鼠炎症性肠病的疗效研究

目的探讨粒-巨噬细胞集落刺激因子(granulocyte macrophage colony-stimulating factor,GMCSF)对于5%2、4、6-三硝基苯磺酸(TNBS)诱导的结肠炎BALB/c小鼠的治疗作用。方法应用TNBS灌肠建立BALB/c小鼠结肠炎模型。结肠炎小鼠随机分为2组,在造模后3h,分别给予GM-CSF(100μg/kg)或磷酸盐缓冲液(PBS)皮下注射进行治疗干预,连续6日。每日观察小鼠体重,腹泻及便血情况,记录第1,3,5日的疾病活动指数(disease activity index,DAI)评分。造模后第5天处死小鼠,获取结肠组织,根据粘连、溃疡及炎症情况计大体损伤评分。应用H-E染色判定结肠病理组织评分;获取结肠组织匀浆,应用ELISA方法检测炎症细胞因子TNF-α的表达。结果 GM-CSF的治疗明显提高了小鼠的生存率,与PBS对照组相比,治疗组小鼠第3、5日的DAI评分、大体损伤评分及病理组织评分明显降低,GM-CSF组小鼠结肠组织中TNF-α的表达水平下调。结论 GM-CSF治疗减轻了小鼠的结肠炎症。     

冻融胚胎移植周期中粒细胞集落刺激因子宫腔灌注对薄型子宫内膜患者的疗效观察

目的观察薄型子宫内膜患者使用粒细胞集落刺激因子宫腔灌注后内膜形态、厚度的变化,分析在冻融胚胎移植周期中薄型子宫内膜患者使用粒细胞集落刺激因子宫腔灌注后胚胎种植率和临床妊娠率的变化。方法回顾性分析2016年12月-2018年12月在我院行冻融胚胎移植的186例薄型子宫内膜患者的临床资料,根据冻融胚胎移植前内膜准备时是否行粒细胞落刺激因子宫腔灌注治疗将患者分为研究组和对照组;内膜准备时在常规治疗基础上,加用粒细胞集落刺激因子宫腔灌注治疗的为研究组,共95例;内膜准备时只用常规治疗的为对照组,共91例。比较移植前子宫内膜准备时两组患者治疗前后子宫内膜形态和厚度的变化,对比胚胎移植后两组患者的胚胎种植率、临床妊娠率、流产率以及异位妊娠率。结果研究组治疗前后内膜形态比较、对照组治疗前后内膜形态比较、治疗后研究组与对照组内膜形态比较,差异无统计学意义(P>0.05);研究组治疗后内膜厚度较治疗前有增加、对照组治疗后内膜厚度较治疗前有增加、治疗后研究组内膜比对照组内膜厚,但差异无统计学意义(P>0.05);胚胎移植后研究组的胚胎种植率(24.85%)明显高于对照组(14.91%),差异有统计学意义(P<0.05),研究组临床妊娠率(41.05%)略高于对照组(30.77%),但差异无统计学意义(P>0.05)。结论未见粒细胞集落刺激因子宫腔灌注促使薄型子宫内膜患者内膜的形态和厚度发生有意义的变化;在冻融胚胎移植周期中薄型子宫内膜患者使用粒细胞集落刺激因子宫腔灌注后胚胎种植率高于常规治疗的患者,但临床妊娠率相差不大;提示粒细胞集落刺激因子宫腔灌注可以提高胚胎种植率,但作用机制可能并不是通过改变其子宫内膜形态或是增加其内膜厚度。     

粒细胞集落刺激因子联合缺血后适应治疗急性心肌梗死的实验研究

 目的：探讨粒细胞集落刺激因子（G-CSF）联合缺血后适应（IP）对实验兔急性心肌梗死的心脏保护作用。方法：将兔结扎左室支（LVA）后分为4组：缺血再灌注模型组（IR组,松结后直接再灌注）、G-CSF组（松结并于术后12 h开始使用G-CSF皮下注射)、IP组（予以IP行改良再灌注）和G-CSF + IP组（联合应用IP及G-CSF干预）。观察各组术中心电图ST段变化,检测手术前后血常规和心肌钙蛋白I（cTnI）,术后4周行超声心动图检查,测定各组梗死心肌的面积、组织病理结构和细胞凋亡。结果：IP组术中ST段回落快于模型组;G-CSF处理后,实验动物血常规中白细胞明显增高;IP组、G-CSF组及联合治疗组术后7 d的cTnI明显低于模型组;术后4周超声心动图结果显示,IP组、G-CSF组及联合治疗组指标好于模型组,联合治疗组最佳;IP组、G-CSF组及联合治疗组梗死心肌面积均明显低于模型组,梗死周边细胞凋亡程度低、血管密度高,联合治疗组最好。结论：G-CSF联合IP可有效减轻心肌梗死后急性期的再灌注损伤,有利于恢复期的心脏修复。     

Mechanisms for T cell tolerance induced with granulocyte colony-stimulating factor

Granulocyte colony-stimulating factor (G-CSF) has been widely accepted as a mediator of T cell tolerance. The immune modulatory effect of G-CSF on T cells is believed to be mediated exclusively through other effector cells, such as monocytes, tolerogenic dendritic cells (DC), and myeloid-derived suppressor cells. Recent advances confirmed the direct effects of G-CSF in inducing immune tolerance of T cells through the G-CSF-G-CSF receptor pathway and related molecular mechanisms. This review aims to summarize the findings associated with the direct and indirect mechanisms for T cell tolerance induced with G-CSF. The role of G-CSF in preventing graft-versus-host disease (GVHD) and in treating autoimmune diseases (ADs) is also discussed. It is conceivable that G-CSF and immune cell compositions, such as tolerogenic DC and CD4⁺CD25⁺Foxp3⁺ T cells, modulated by G-CSF could become an integral part of the immunomodulatory therapies against GVHD and ADs in the future.     

Mechanisms of hemostimulating effect of granulocytic colony-stimulating factor during cytostatic treatment

Original Russian-made preparation of human recombinant granulocytic colony-stimulating factor possessesin vitro activity similar to that of standard preparation, activatesin vivo production of humoral factors by adherent cells of the hemopoiesis-inducing microenvironment, and stimulates hemopoiesis in the bone marrow at various levels. Translated fromByullenten' Eksperimental'noi Biologii i Meditsiny, Vol. 128, No. 8, pp. 194–199, August, 1999     

重组人血清白蛋白/粒细胞刺激因子融合蛋白的药效学、药理学和毒理学研究

目的对重组人血清白蛋白/粒细胞刺激因子融合蛋白(rHSA/GCSF)开展药效学、药理学和毒理学动物评价研究,以确认其安全性和有效性。方法①对rHSA/GCSF开展的药效学研究内容主要有:以恒河猴骨髓细胞为研究系统,在体外条件下培养骨髓粒细胞-巨噬细胞集落(CFU-GM),计数不同浓度的rHSA/GCSF对CFU-GM数的影响的体外药效学评价;以小鼠~(60)Coγ照射、氟尿嘧啶注射液所致的鼠白细胞低下和对~(60)Coγ射线照射致食蟹猴白细胞低下的治疗作用。②药理学研究观察了rHSA/GCSF对小鼠中枢神经系统和狗呼吸及心血管系统功能的影响。③毒理学研究考察了rHSA/GCSF静脉和皮下给予小鼠、食蟹猴的急性毒性反应,以及长期和反复给药对大鼠和食蟹猴的安全性做出评价。④rHSA/GCSF的药代/毒代试验则是对~(125)I-rHSA/GCSF不同剂量单次皮下注射给予小鼠、大鼠后,rHSA/GCSF在各器官的分布、总放射性和TCA沉淀放射性的动力学参数测定,以及对不同剂量rHSA/GCSF连续给药食蟹猴后的血浆药物浓度及代谢动力学参数开展了考察。结果①rHSA/GCSF可以使骨髓的粒白细胞系(粒系)增生活跃,骨髓粒系造血细胞及成熟中性粒细胞均明显增多;对氟尿嘧啶所致小鼠白细胞减少症有明显的治疗作用,在使用化疗药早期,可以减缓白细胞的降低,特别是可以使粒白细胞提前恢复到正常水平;rHSA/GCSF可显著缩短食蟹猴白细胞减少症放疗模型产生的白细胞低下持续时间,使外周血白细胞加速恢复,尤其以中性粒细胞的增加为主,同时对红细胞和血小板的影响不大。②从获得的PD/PK数据t_(1/2)来看:rHSA/GCSF半衰期平均值约为38.6 h;单次皮下注射rHSA/GCSF,在500、1500、3000μg/kg剂量范围内对小鼠中枢神经系统无影响,与戊巴比妥钠无协同作用;单次皮下注射rHSA/GCSF在50、200μg/kg剂量范围内对狗呼吸和心血管系统无明显影响。实验表明rHSA/GCSF单次皮下和静脉注射给予小鼠的最大耐受量(MTD)≥37.5 mg/kg,单次皮下注射给予食蟹猴的最大耐受剂量为11.6 mg/kg:长期反复给药对大鼠的基本安全剂量为300μg/kg,对食蟹猴则是≥150μg/kg。结论 rHSA/GCSF融合蛋白每4天给药1次,对放、化疗所致的动物外周血白细胞减少症具有治疗作用,与市售常规rhGCsF每天注射1次相比具有长效作用。所获得的大量药效学、药代动力学、毒理学、毒代动力学的试验数据可供正在开展的临床试验研究参考,并具有很好的指导意义。     

Macrophage-granulocyte colony-stimulating factor and fibroblast growth factor in the blood of patients with hyperlipoproteinemia

Solid-phase ELISA revealed increased content of macrophage-granulocyte colony-stimulating factor and basic fibroblast growth factor in the serum of hyperlipoproteinemic patients.     

Effects of granulocyte colony-stimulating factor on the kinetics of inflammatory cells in the peripheral blood and pulmonary lesions during the development of bleomycin-induced lung injury in rats

We evaluated the effects of granulocyte colony-stimulating factor (G-CSF) on the kinetics of inflammatory cells during the development of inflammation in bleomycin (BLM)-induced lung injury. G-CSF (100 microg/kg/day, s.c.) was administered to rats treated with or without BLM (2 mg/200 microl, intratracheally) for up to 14 days (Day 14) immediately after BLM treatment. In the BLM + G-CSF group, the lung injury score increased on Days 1 and 14, and the score of lung fibrosis on Day 14, respectively. Except for neutrophils, there were no effects of G-CSF on the number of inflammatory cells both in the peripheral blood and in the lung in both BLM-treated and -untreated rats at the acute inflammatory phase. In the G-CSF-treated groups, the number of neutrophil counts in the peripheral blood drastically increased on Day 1, temporally decreased on Day 3, and increased again on Days 7 and 14. The number of neutrophils in the lung markedly increased on Day 1 and then remained at a plateau level until Day 14. The neutrophil alkaline phosphatase score in the lung commenced to increase on Day 1, reached the maximal level on Day 7, and then remained at a plateau level until Day 14. Correlations between the numbers of neutrophils in the lung and the peripheral blood or the lung lesion score were only observed on Day 14. These findings suggest that the exacerbating effect of G-CSF on the lung injury coincided with the increase in the number of alkaline phosphatase-positive neutrophils infiltrating in the pulmonary lesion at the acute inflammatory phase and it lasted to the fibrogenic phase. The exacerbating effect of G-CSF on the severe BLM-induced lung injury seems to be related not only to the pulmonary accumulation of activated neutrophils but also to the severity of lung injury caused by the direct effects of BLM.     

艾滋病抗病毒治疗48例死亡患者相关因素分析

目的分析本院艾滋病抗病毒治疗死亡患者的相关因素。方法利用国家统一使用的DataFax抗病毒治疗信息系统所收集的数据资料,对抗病毒治疗死亡患者的相关情况进行分析。结果共有315例艾滋病患者接受治疗,有48例抗病毒治疗患者死亡。治疗前所有患者出现艾滋病临床Ⅲ期或Ⅳ期表现,CD4细胞最小值为4个/μl,最大值为263个/μl,平均为36.4个/μl。接受治疗时间最短1d,最长2年半,其中39例的治疗时间小于3个月。死于艾滋病相关性疾病42例,其中9例系统治疗无效死亡,11例治疗过程中出现机会性感染,放弃治疗在家中死亡;22例在停药后0.5～8月出现艾滋病相关疾病死亡。自杀4例,死亡原因不明2例。结论艾滋病抗病毒治疗患者的主要死亡原因是艾滋病相关疾病,依从性困难是患者死亡的另一个原因,经济困难、歧视和药物副作用与病人死亡有直接关系。     

高血压病血清巨噬细胞集落刺激因子受体的变化及其意义

目的：探讨高血压病巨噬细胞集落刺激因子受体（M-CSFR）的改变及其临床意义。方法：用多克隆双抗体夹心ABC-ELISA法测定58例高血压病患者和30例正常对照组的血清M-CSFR。结果：高血压病组血清M-CSFR水平明显高于对照组（P<0.01）、1级高血压组M-CSFR明显低于3级高血压组（P<0.01）。相关分析发现：血清M-CSFR浓度与收缩压、舒张压和平均动脉压呈正相关（r=0.65、r=0.68、r=0.73，P均<0.01）。结论：高血压病患者M-CSFR水平升高在血管内膜损伤和动脉粥样硬化过程中起重要作用。     

A combination of stem cell factor and granulocyte colony-stimulating factor enhances the growth of human progenitor B cells supported by murine stromal cell line MS-5

We have developed a long-term culture system using the murine bone marrow stromal cells MS-5 to support the growth of progenitor B cells with CD34^–, CD10⁺, CD19⁺, and cytoplasmic μ chain (Cμ)-negative surface phenotype from human CD34⁺ cells purified from umbilical cord blood (CB). When 10³ CB CD34⁺ cells/well were seeded on MS-5 stromal cells at the beginning of culture in the absence of exogenously added cytokines, progenitor B cells first appeared after 14 days, and the maximal cell production was achieved during the 6th week of culture. Intriguingly, the addition of recombinant human stem cell factor (rhSCF) and granulocyte colony-stimulating factor (rhG-CSF), but not rhIL-7, strikingly enhanced the growth of progenitor B cells from CB CD34⁺ population cultured on MS-5 stromal cells. The culture of progenitor B cells could be maintained until the 6th week of culture when some cells were revealed to have a Cμ⁺ phenotype, and a small number of cells had immunoglobulin μ chain on their cell surface in the presence of both rhSCF and rhG-CSF. When CD34⁺ cells were cultured physically separated from the stromal layer by membrane, supportive effects of MS-5 stromal cells for the growth of progenitor B cells were not observed. These results suggest that the present culture system could generate progenitor B cells to proliferate from CB CD34⁺ cells, that some of these progenitor B cells could differentiate into immature B cells in conjunction with rhSCF and rhG-CSF, and that a species-cross-reactive membrane-bound factor(s), which stimulates early human B lymphopoiesis, may exist in MS-5 stromal cells. Further studies are required to investigate the mechanism how rhG-CSF acts on progenitor B cells to allow their proliferation and differentiation.