体内表达谷胱甘肽过氧化物酶-7型基因对骨髓抑制的防护作用研究

 目的 探讨体内表达谷胱甘肽过氧化物酶-7 基因（Gpx-7）对 5-氟尿嘧啶（5-Fu）引发小鼠骨髓抑制的防护作用。方法 以尾静脉注射5-Fu（250 mg/kg）的方法建立小鼠骨髓抑制和再生模型。5-Fu 注射后第 1 天，通过电穿孔转染法将 pcDNA3.1- Gpx7 重组质粒 50 μg 注入小鼠胫前肌（实验组），以 pcDNA3.1 空质粒载体 50 μg 电穿孔转染作为对照组，每组各 30 只小鼠。 5-Fu 注射前和注射后 3、7、11、14 d，2 组各断颈处死 6 只小鼠，计数小鼠外周血白细胞数、血小板数和单条腿骨髓细胞总数；然后分别取 5-Fu 注射后 7、14 d 断颈处死小鼠各 3 只进行骨髓细胞集落形成试验；并分别取 5-Fu 注射前和注射后 3、7、11、14 d 断颈处死的小鼠各 1 只制备完整大腿骨石蜡组织切片，观察骨髓组织形态学的变化。结果 注射 5-Fu 后 11 和 14 d，实验组外周血白细胞数分别为（3917 ± 733）和（6857 ± 1878）/μl，均明显高于同时点对照组[分别为（2683 ± 920）和（4017 ± 1011）/μl，均 P < 0.05）]；注射 5-Fu 后 11 d，实验组外周血小板数为（150.8 ± 64.3）万/μl，明显高于同时点对照组[（63.0 ± 60.3）万/μl，P < 0.05）]；注射 5-Fu 后不同时间 2 组间单条腿骨髓细胞总数差异无统计学意义。注射 5-Fu 后 7、14 d，2 组之间骨髓细胞集落形成数差异无统计学意义。 实验组注射 5-Fu 后 11 d 小鼠的骨髓组织形态恢复程度与对照组注射 5-Fu 后 14 d 小鼠类似；注射 5-Fu 后 14 d 小鼠的骨髓组织形态接近注射前。 结论 体内表达 Gpx-7 基因可以促进被化疗药物 5-Fu 抑制的小鼠骨髓再生。     

体内表达谷胱甘肽过氧化物酶-7基因对骨髓抑制的防护作用研究

目的 探讨体内表达谷胱甘肽过氧化物酶-7基因(Gpx-7)对5-氟尿嘧啶(5-Fu)引发小鼠骨髓抑制的防护作用.方法 以尾静脉注射5.Fu(250 mg/kg)的方法建立小鼠骨髓抑制和再生模型.5-Fu注射后第1天,通过电穿孔转染法将pcDNA3.1-Gpx7重组质粒50 μg注入小鼠胫前肌(实验组),以pcDNA3.1空质粒载体50μg电穿孔转染作为对照组,每组各30只小鼠.5-Fu注射前和注射后3、7、11、14 d,2组各断颈处死6只小鼠,计数小鼠外周血白细胞数、血小板数和单条腿骨髓细胞总数;然后分别取5-Fu注射后7、14 d断颈处死小鼠各3只进行骨髓细胞集落形成试验;并分别取5-Fu注射前和注射后3、7、11、14d断颈处死的小鼠各1只制备完整大腿骨石蜡组织切片,观察骨髓组织形态学的变化.结果 注射5-Fu后11和14 d,实验组外周血白细胞数分别为(3917±733)和(6857±1878)/μl,均明显高于同时点对照组[分别为(2683±920)和(4017±1011)/μl,均P<0.05)];注射5-Fu后11 d,实验组外周血小板数为(150.8±64.3)万/μl,明显高于同时点对照组[(63.0±60.3)万/μl,P<0.05)];注射5-Fu后不同时间2组间单条腿骨髓细胞总数差异无统计学意义.注射5-Fu后7、14 d,2组之间骨髓细胞集落形成数差异无统计学意义.实验组注射5-Fu后11 d小鼠的骨髓组织形态恢复程度与对照组注射5-Fu后14 d小鼠类似:注射5-Fu后14 d小鼠的骨髓组织形态接近注射前.结论 体内表达Gpx-7基因可以促进被化疗药物5-Fu抑制的小鼠骨髓再生.     

红景天多糖对骨髓抑制贫血小鼠外周血及骨髓细胞周期的影响

目的观察红景天多糖对骨髓抑制贫血小鼠外周血及骨髓细胞周期的影响,并探讨其造血调控作用机制。方法用全自动血细胞分析仪、白细胞计数法、流式细胞术（FCM）分别检测红景天多糖对骨髓抑制贫血小鼠外周血、骨髓有核细胞、骨髓细胞周期的影响。结果中剂量和高剂量红景天多糖能明显升高外周血白细胞（WBC）、红细胞（RBC）、血红蛋白（Hb）及骨髓有核细胞（BMCs）数,但对血小板的作用不明显。高剂量组可促进骨髓G0／G1期细胞向S期细胞以及S期细胞向G2／M期细胞的转化,增殖指数（PI）也明显升高。结论红景天多糖可能通过促使骨髓抑制贫血小鼠骨髓细胞通过G1期的限制点,进入细胞增殖周期,加速骨髓G0／G1期细胞向S期细胞、S期细胞向G2／M期细胞的转化,促进骨髓造血细胞增殖,提高外周血象,促进骨髓造血功能的恢复。     

家蚕表达的重组hM-CSF对化疗损伤小鼠造血功能的恢复

目的 研究重组人巨噬细胞集落刺激因子（ｒｈＭ－ＣＳＦ）和对化疗损伤小鼠骨髓造血功能恢复的作用。方法 用环磷酰胺１００ｍｇ／ｋｇ连续３天给予小鼠腹腔注射,造成化疗损伤模型。继以不同剂量（０．１￣２．０μｇ／天）的ｒｈＭ－ＣＳＦ连续７天给小鼠腹腔注射治疗。在化疗后第５,８,１１,１４天进行外周血白细胞总数及中性粒细胞、单核细胞和淋巴细胞计数;并在化疗后第７天进行骨髓ＣＦＵ－ＧＭ的集落培养。数据均采用＾     

淫羊藿苷对化疗后小鼠骨髓和细胞免疫抑制作用的影响

目的:观察淫羊藿苷(ICA)对环磷酰胺(Cy)所致小鼠骨髓和免疫抑制作用的影响,探讨ICA促进化疗后小鼠造血功能和免疫功能的作用及机制。方法:将小鼠随机分为6组:正常对照组,模型组,阳性对照组,ICA高、中、低剂量组。除正常对照组小鼠外,其余各组小鼠均腹腔注射Cy(200mg/kg)。第2天开始,对ICA高、中、低剂量的实验组小鼠灌胃不同剂量的ICA(150、80、40mg/kg.d),阳性对照组小鼠尾静脉注射参芪扶正注射液(1mL/d),模型对照组给予等量生理盐水,连续给予10d。经HE染色后观察小鼠胸腺组织结构的变化。用MTT比色法检测脾淋巴细胞的增殖率。用乳酸脱氢酶(LDH)法检测腹腔巨噬细胞对肿瘤细胞的杀伤能力。用ELISA试剂盒检测混合细胞培养上清中TNF-α和IL-12的含量。用全自动血液分析仪检测外周血红细胞、白细胞和血小板数量的变化;外周血和股骨骨髓涂片经瑞氏-吉姆萨染色后,光镜下计数单根股骨骨髓细胞(BMC)的数量。结果:ICA具有保护小鼠胸腺、骨髓免受Cy损伤的作用。不同剂量的ICA组小鼠的脾淋巴细胞增殖能力增加,巨噬细胞吞噬能力和分泌细胞因子的能力均增强,外周血红细胞、白细胞和血小板数量均明显上升。结论:ICA可逆转Cy化疗后小鼠骨髓造血和免疫功能的抑制状况。     

沙棘汁对低白细胞模型小鼠白细胞和骨髓造血功能的调节作用

目的： 研究沙棘汁提升低白细胞模型小鼠白细胞数量和促进骨髓细胞增殖功能的作用。 方法： 应用环磷酰胺（CTX）（100 mg/kg）给小鼠腹腔注射，复制白细胞减少症模型，分组治疗处理后，于不同时段计数外周血白细胞总数并分类，计数骨髓有核粒细胞及其分裂指数，以评估沙棘汁提升白细胞及增强骨髓造血的能力。 结果： 低、中、高剂量沙棘汁均可显著提升模型小鼠的外周血白细胞数，并增加骨髓有核粒细胞数和促进有核细胞的分裂，效果均优于碳酸锂组；在疗效与剂量关系上呈明显的量效关系。 结论： 沙棘汁可明显增加低白细胞小鼠模型的外周血白细胞数量及骨髓细胞增殖功能。     

IL—3基因疗法对化疗造血损伤的恢复作用

白细胞介素３（ＩＬ－３）是一种具有很强造血调控作用的细胞因子。本实验观察了成纤维细胞介导的ＩＬ－３基因疗法对大剂量环磷酰胺体内注射后造血功能损伤小鼠的恢复作用。结果表明：接爱ＩＬ－３基因疗法的实验小鼠外周血白细胞和血小板数量降低程度减弱，回升速度加快，其脾脏和骨髓ＣＦＵ－ＧＭ、ＣＦＵ－Ｅ、ＣＦＵ－ＭＫ、ＣＦＵ－Ｓ水平显著地高于对照小鼠，可见成纤维细胞介导的ＩＬ－３基因疗法可显著地降低大剂量化疗后造     

重组人白细胞介素—1β对化疗小鼠巨核系造血及存活率的影响

目的：观察重组人白细胞介素１β（ｒｈＩＬ－１β），对用细胞毒化疗药物后，机体巨核系造血功能恢复及存活率的影响。结果：①给予不同剂量ｒｈＩＬ－１β（３×１０３～３×１０５Ｕ／鼠）后小鼠腹腔注射半数致死量环磷酰胺，外周血血小板的恢复明显加快，其中以３×１０５Ｕ剂量组效果最佳；注射后５ｄ骨髓ＢＦＵ－Ｍｅｇ含量随给药剂量加大而增加。②给予不同剂量ｒｈＩＬ－１β（３×１０３～３×１０５Ｕ／鼠）后小鼠腹腔注射致死量环磷酰胺，３０ｄ存活率各用药组较对照组均有提高，其中以３×１０５Ｕ组结果最佳（Ｐ＜００５）。结论：ｒｈＩＬ－１β能促进注射半致死量环磷酰胺的小鼠巨核系造血功能恢复，并提高注射致死量环磷酰胺小鼠的３０ｄ存活率。     

IL－3基因疗法对化疗造血损伤的恢复作用

白细胞介素３（ＩＬ－３）是一种具有很强造血调控作用的细胞因子，本实验观察了成纤维细胞介导的ＩＬ－３基因疗法对大剂量环磷酰胺体内注射后造血功能损伤小鼠的恢复作用。结果表明：接受ＩＬ－３基因疗法的实验小鼠外周血白细胞和血小板数量降低程度减弱，回升速度加快，其脾脏和骨髓ＣＦＵ－ＧＭ、ＣＦＵ－Ｅ、ＣＦＵ－ＭＫ、ＣＦＵ－Ｓ水平显著地高于对照小鼠，可见成纤维细胞介导的ＩＬ－３基因疗法可显著地降低大剂量化疗后造血损伤程度，并能显著地促进受损的造血功能尽快恢复，提示若将ＩＬ－３基因疗法与大剂量化疗联合应用将提高肿瘤治疗的效果。     

国产基因重组人粒细胞集落刺激因子的药效学研究

本文通过国产基因重组人粒细胞集落刺激因子和进口(日产)人粒细胞集落刺激因子,对C_(57)小鼠经环磷酰胺造成的骨髓抑制和经60Co辐射所致的造血功能抑制的保护作用比较研究,结果表明,国产与进口制剂对C_(57)小鼠环磷酰胺引起的白细胞降低模型有升高白细胞,促进骨髓细胞集落形成(CFU—GM),脾结节形成增加,和脾脏系数增大作用,且两种制剂在剂量100μg/kg对作用无明显差异(P>0.05),两种rhG-CSF对~(60)Co辐射引起的骨髓抑制均能促进白细胞增加,提高骨     

人外周血γδT 细胞免疫治疗裸鼠皮下人肝癌细胞移植瘤的体内实验研究

目的:研究不同剂量的人外周血γδT 细胞对裸鼠人肝癌细胞(SMMC-7721)移植瘤模型的免疫治疗作用。方法: 用人肝癌细胞株SMMC-7721 接种BALB/ c 裸鼠皮下,建立肝癌裸鼠模型。于从健康人外周血中提取单核细胞,体外特异性扩增γδT 细胞。将已建立的肝癌裸鼠模型随机分为5 组,阳性对照组为5-氟尿嘧啶(5-Fu),阴性对照组为生理盐水,治疗组用不同剂量的γδT 细胞 分别注入裸鼠尾静脉,阳性对照组用5-Fu 裸鼠腹腔注射,阴性对照组用生理盐水裸鼠尾静脉注射。观察不同剂量的γδT 细胞对肿瘤的抑制效果,包括治疗前后的体重、食物摄取量及生长状况等,并与阳性对照组和阴性对照组比较肿瘤体积(TV)、相对肿瘤体积(RTV)和相对肿瘤增殖率[T/ C(%)]变化。结果:不同剂量的γδT 细胞对裸鼠移植瘤的生长有不同程度的抑制,RTV 与生理盐水阴性对照组比较差异有统计学意义(P<0.05);与5-Fu 阳性对照组比较,TV 增长明显低于5-Fu 阳性对照组,RTV 各剂量组抑制程度相似,均略高于5-Fu 阳性对照组,T/ C(%)各剂量组比5-Fu 对照组的相对肿瘤增殖率稍低,但无显著性差异。结论:人外周血γδT 细胞对肝癌裸鼠移植瘤具有显著的抑瘤作用,为建立肝癌免疫治疗新方法提供实验依据。     

BCG 感染小鼠模型中产IL鄄35 单核细胞的检测及其意义

目的:检测BCG 感染小鼠模型外周血和肺组织中产IL-35 单核细胞,分析其在结核病免疫机制及病理、病程中发挥的作用,探讨其临床意义。方法:用BCG 感染C57BL/6 小鼠,于不同时间处死小鼠取肺组织进行载菌量和组织病理学分析,取外周血与肺组织,并采用流式细胞术检测IL-35 两个亚基P35 与EBI3 在单核细胞中的表达,分析IL-35 与小鼠载菌量的相关性。结果:BCG 感染小鼠外周血和肺组织中不同时间点单核细胞P35、EBI3 表达量明显高于对照组,且实验组外周血在感染4 周时显著增加,肺组织表达EBI3 在4、8 周有显著升高趋势,且P35 与EBI3 的表达呈正相关,肺组织IL-35 的表达与荷菌量呈负相关。外周血和肺部IL-35 表达水平在2、4 周均有显著增加,而第8 周时仅肺部升高明显。结论:BCG 感染小鼠单核细胞中IL-35 高表达,可能参与抗结核免疫调节作用。     

Interferon-�� increases monocyte migration via platelet�Cmonocyte interaction in murine intestinal microvessels

The aim of this study was to investigate the effect of interferon (IFN)‐α on recruitment of platelets and monocytes within the murine small intestinal venular endothelium. Monocytes were isolated from bone marrow of C57B6 mice. Platelets were collected from murine blood. Rolling and adhesion to submucosal microvessels in the small intestine were examined under an intravital fluorescence microscope after injection of fluorescein‐labelled monocytes or platelets. In some mice, IFN‐α (5 × 10⁵U/kg) was administered intraperitoneally. After treatment with an antibody against P‐selectin, changes in monocyte and platelet migration were also investigated. Changes in monocyte migration under the condition of thrombocytopenia were also investigated. Platelets and monocytes interacted with murine intestinal microvessels, although only few platelets and monocytes showed migration behaviour. Intraperitoneal injection of IFN‐α enhanced the migration of both platelets and monocytes in the intestinal microvessels. Pretreatment with anti‐P‐selectin attenuated the increase in migration of platelets and monocytes induced by administration of IFN‐α. Thrombocytopenia decreased the rolling ratio of monocytes, suggesting that the effect of IFN‐α on migration was P‐selectin‐dependent, derived from both the endothelium of microvessels and platelets. The results of this study suggest that IFN‐α acts as a potent proinflammatory agent via its stimulatory effect on the endothelium–platelet–monocyte interaction in intestinal microvessels by a P‐selectin‐dependent mechanism.     

Accelerated recovery from gamma-irradiation-induced leukopenia in mice by the biscoclaurine alkaloid, Cepharanthin--comparison with recombinant human granulocyte colony-stimulating factor.

The effect of a biscoclaurine alkaloid drug, Cepharanthin (CE), on recovery from severe leukopenia induced by whole body gamma-irradiation at a dose of 3 Gy, was compared with that of recombinant human granulocyte colony-stimulating factor (rhG-CSF). Daily intraperitoneal administration of 100 micrograms CE into the irradiated mice significantly prevented decrease of leukocyte numbers in the peripheral blood and accelerated recovery from leukopenia to normal. The activity of CE was somewhat weaker than that of 20,000 units of rhG-CSF when administered daily subcutaneously. In cell composition of peripheral leukocytes, CE increased the numbers of polymorphonuclear (PMN) cells and lymphocytes compared with those of the irradiated controls. Stronger change in PMN cell numbers than with CE was induced by administration of rhG-CSF. No significant increment of the activity to form colonies in spleen (colony-forming unit in spleen; CFU-S) was observed in bone marrow cells from irradiated donor mice administered CE, though CE induced CSF production into sera. Administration of rhG-CSF stimulated the CFU-S activity. These results indicate that CE increased recruitment of PMN cells from the marginal pool rather than the stimulation of pluripotent stem cells in the bone marrow.     

Effect of irradiation,cyclophosphamide, and etoposide (VP-16) on number of peripheral blood and peritoneal leukocytes in mice under normal conditions and during acute inflammatory reaction

In order to develop a suitable model for studying the role of granulocytes and monocytes in resistance against pathogenic microorganisms, we investigated the effect of irradiation and cytostatic treatment (cyclophosphamide and VP-16) on the number of both peripheral blood and peritoneal leukocytes in male Swiss mice. Irradiation and cyclophosphamide treatment severely decreased the number of both granulocytes and monocytes in peripheral blood, whereas VP-16 only lowered the number of blood monocytes to a significant degree and had little effect on the number of blood granulocytes or lymphocytes. When normal mice were injected intraperitoneally with newborn calf serum (NBCS) the number of peritoneal granulocytes rose about 100-fold within 6 h. In irradiated and cyclophosphamide-treated mice, this influx of granulocytes into the peritoneal cavity was virtually eliminated, as was the concomitant increase in the number of blood granulocytes; in VP-16-treated mice, on the other hand, the number of peripheral blood and peritoneal granulocytes increased to the same degree as in normal mice. An increase in the number of peripheral blood monocytes and peritoneal macrophages occurred 24–48 h after injection of NBCS in normal mice. This increase was significantly impaired by irradiation as well as by treatment with cyclophosphamide or VP-16.     

Enhancement of host defense by Y-25510, (+-)-3-[4-(2-dimethylamino-1-methylethoxy)phenyl]-1H-pyrazolo[3,4 -b] pyridine-1-acetic acid, a novel synthetic compound. A comparison with recombinant human granulocyte colony-stimulating factor in 5-fluorouracil-treated mice.

The effect of a novel synthetic compound, Y-25510, (+-)-3-[4-(2-dimethylamino-1-methylethoxy)phenyl]-1H-pyrazolo[3,4 -b] pyridine-1-acetic acid, on recovery from long-lasting leukopenia induced by 5-fluorouracil was compared with that of recombinant human granulocyte colony-stimulating factor (rhG-CSF). When mice were administered i.p. with 5-FU (200 mg/kg) on days 0 and 7, intravenous administration of Y-25510 (100 and 1000 micrograms/kg) prevented the decrease in the peripheral leukocyte and neutrophil number and accelerated the recovery from leukopenia. Subcutaneous administration of rhG-CSF (50 micrograms/kg) did not prevent leukopenia but accelerated the recovery from leukopenia. In particular, peripheral neutrophil number increased over a normal level. The administration of Y-25510 (10, 100 and 1000 micrograms/kg) restored the decrease in the number of bone marrow cells, spleen cells, lymphocytes, neutrophils and monocytes. The administration of rhG-CSF (50 micrograms/kg) restored the decrease in the number of bone marrow cells, spleen cells, and neutrophils but not that of lymphocytes and monocytes. In fractions of bone marrow cells on day 21, the administration of Y-25510 (1000 micrograms/kg) showed a tendency of restoring the decrease in neutrophil number. In conclusion, the administration of Y-25510 prevented leukopenia and accelerated the recovery from leukopenia in the 5-FU-treated mice. It is suggested that the mechanism of the restorative action of Y-25510 is different from that of rhG-CSF. In a number of immature bone marrow cells Y-25510 has a potent stimulatory effect on the recovery from the decrease in number of hematopoietic cells, keeping a balance in number of each blood cell.     

rhIL-11和rhG-CSF联合应用对小鼠外周血T细胞亚群比例影响

为观察rhIL 11联合rhG CSF动员小鼠外周造血干细胞时 ,外周血T细胞亚群的变化 ,使用rhIL 11联合rhG CSF动员C5 7BL/ 6小鼠外周造血干细胞 ,观察用药不同时间小鼠外周血白细胞变化 ,同时通过流式细胞仪测定外周血T细胞亚群的变化。结果发现 ,小鼠注射rhIL 11后 ,外周血WBC明显升高 (P =0 0 0 3) ;单独注射rhG CSF或联用rhIL 11后 ,外周血WBC于第 5天达峰值 ,分别由用药前的 (7 5 3± 1 6 5 )× 10 9/L、 (7 2 7± 1 4 8)× 10 9/L上升至 (2 7 12± 1 84 )× 10 9/L、 (2 8 98± 3 13)× 10 9/L ,均显著高于对照组 (P值均为 0 0 0 1) ,而rhG CSF组与联合组之间无显著性差异 (P >0 0 5 )。CD3+ 细胞比例在用药组均高于正常对照组 (P值均 <0 0 5 ) ,rhIL 11组CD3+ 细胞及CD4 + 细胞比例逐渐增高 ,于动员的第 5天达峰值 (P值均为 0 0 0 1) ,CD8+ 细胞的比例没有明显变化 (P >0 0 5 ) ;在rhG CSF组CD3+ 细胞也有明显增高 ,并于动员第五天达峰值 (P =0 0 0 1) ,CD4 + 细胞的比例没有明显变化 (P值均 >0 0 5 ) ,但CD8+ 细胞的比例明显升高 (P =0 0 0 1) ;在联合组中CD3+ 、CD4 + 、CD8+ 细胞的比例变化均有明显升高。结果证明 ,rhIL 11联合rhG CSF能明显提高外周血WBC ,增加CD3+ 细胞比例 ,同时rhIL 11     

养正合剂和粒细胞集落刺激因子对化疗后骨髓功能的影响

目的:探究养正合剂和粒细胞集落刺激因子对化疗后骨髓功能的影响.方法:50只健康雌性C57BL/6小鼠以环磷酰胺(CTX)致骨髓功能抑制模型后随机分为空白组、模型组、rhG-CSF组、养正合剂1组、养正合剂2组,10只/组,连续5 d分别给予生理盐水0.2 ml、生理盐水0.2 ml、重组人粒细胞-巨噬细胞集落刺激因子(...