苦参碱对低剂量照射小鼠骨髓细胞端粒酶活性及细胞凋亡的影响

目的 观察苦参碱(matrine,Ma),对受低剂量照射小鼠骨髓细胞端粒酶活性及细胞凋亡的影响.方法 采用Varine 600直线加速器为照射源,以0.2 Gy/d的小剂量分次照射小鼠,分别于照射5、10、15、20天后的1、3、6、10、15、21天分批活杀小鼠,涂片观察小鼠骨髓细胞的增生程度,采用端粒重复扩增(te...     

Study on the effects of Guiqi Oral Liquid in promoting recovery of hematopoiesis in acute irradiation injured mice

Objective: To explore the effects and possible mechanisms of Guiqi Oral Liquid (GQOL) on the recovery of hematopoiesis in acute irradiation injured mice.Methods: The acute irradiation injured mice were randomly divided into 2 groups: the treated group and the control group, and also a normal control group was set up with 6 mice in it receiving no treatment. After the mice in the former two groups were irradiated by 6.0 Gy⁶⁰Coγ-ray, every one of them was given 0. 4 ml GQOL or saline in equal volume through a gastric tube twice a day for 14 days. On the 4th, 8th and 14th day after irradiation, the bone marrow mononuclear cells (BMMNC) and megakaryocytes in bone marrow tissues of the mice were counted, the proportion of hematopoietic tissues (by area) was measured, and the expression of adhesion molecules, CD44 and CD54, in bone marrow were estimated by immunochemistry. The colony forming unit of spleen (CFU-S) in the mice were counted on the 8th day after irradiation.Results: On the 4th, 8th, 14th day after irradiation, the count of BMMNC and megakaryocyte, and the proportion of hematopoietic tissues in the treated group were higher than those in the control group (P<0.01 orP<0.05). CD44 and CD54 expression in the treated group were higher than those in the control group on the 4th and 8th day (P<0.01), but near normal on the 14th day (P<0.01). On the 8th day, CFU-S count in the treated group was higher than that in the control group (P<0.01).Conclusion: GQOL can regulate the expression of adhesion molecules, CD44 and CD54, in the bone marrow of the acute irradiation injured mice, which may be one of the mechanisms of GQOL in accelerating the early phase hematopoiesis recovery of mice. Supported by the National Natural Science Foundation of China (No. 39870926)     

放射损伤小鼠骨髓VEGF、CFU-GM、CFU-F的变化及川芎嗪对其影响的研究

目的:探讨放射损伤小鼠骨髓血管内皮细胞生长因子(VEGF)、骨髓粒巨噬细胞集落形成单位(GFU-GM)数和骨髓成纤维细胞集落成单位(CFU-F)的变化意义及川芎嗪对其的影响。方法:健康昆明小鼠经6.0Gy60Coγ照射后,立即喂饲川芎嗪并设对照组和正常组,分别于第3、7、14、21天检测其骨髓VEGF、CFU-GM和CFU-F的变化。结果:照射后第3、7、14 d骨髓VEGF表达水平显著降低,但川芎嗪组高于对照组;随着时间推移骨髓VEGF表达水平逐渐恢复,照射后第21 d川芎嗪组已恢复正常(P>0.05),而照射组仍低于正常组。照射后骨髓CFU-GM和CFU-F明显受到抑制,但川芎嗪组显示了良好的保护作用。结论:放射损伤后小鼠骨髓VEGF的表达变化在造血功能的恢复过程中起重要作用;川芎嗪具有改善骨髓造血功能的作用。     

Enhancing effects of silkworm expressed recombinant human macrophage colony-stimulating factor on hematopoietic recovery of irradiation-injured mice

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Effect of Ligustrazine on Bone Marrow Microenvironment and Signal Transducti on Path in Irradiation Injured Mice

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白细胞介素12对急性放射病小鼠造血系统的影响

目的 研究重组鼠白细胞介素12(rmIL-12)对急性放射病小鼠造血系统的影响.方法 42只BALB/c小鼠均给予6.0Gy(60)γ射线全身照射,随机分为照射对照组、5和20μg/kg的rmIL-12治疗组,治疗组分别于照后1h及此后每3d一次分别腹腔注射5和20 μg/(kg·d)的rmIL-12,共5次,每日2...     

黄芪在外周血造血干细胞移植中应用的初步实验研究

目的为研究中药黄芪对外周血造血干细胞移植术后早期造血功能重建有无促进作用.方法以致死量辐照并行外周血造血干细胞移植的BALB/c小鼠为模型,移植后给予黄芪注射液10g/kg或20g/kg腹腔注射,连续15天,观察4周内外周血象的恢复情况,并对骨髓造血微环境行透射电镜观察.结果黄芪组小鼠外周血白细胞的恢复快于对照组,血小板的恢复与对照组无显著差异,两个黄芪剂量组之间无显著差异;电镜检查发现黄芪组小鼠骨髓中内皮细胞、肉状细胞的损伤轻于对照组,内皮细胞、网状细胞与造血原始细胞之间的接触较对照组紧密.结论黄芪能促进小鼠外周血造血干细胞移植术后早期白细胞的重建,其机理可能是通过保护和改善骨髓造血微环境,改善骨髓基质细胞与造血干细胞之间的接触,并促进G-CSF等内源性细胞因子的分泌.     

rhBMP-2对再生障碍性贫血小鼠骨髓造血恢复的促进作用

探究了人重组骨形态发生蛋白-2(rhBMP-2)对再障小鼠的治疗作用。采用5-FU联合白消安建立小鼠再障模型,通过干预rhBMP-2进行治疗。考察了各实验组小鼠白细胞数、骨髓单核细胞数、体重、存活率、脾系数、粒系-巨系细胞集落(CFU-GM)数以及骨髓单核细胞中CD34⁺细胞比例,进行股骨、脾脏HE切片分析。结果表明,相较于再障对照组,rhBMP-2治疗组小鼠的白细胞数、存活率、CFU-GM集落数及骨髓单核细胞数显著提高,并且骨髓单核细胞中的CD34⁺细胞含量也提高,显著改善脾脏功能和缓解骨髓抑制,证实rhBMP-2能够促进再障小鼠造血损伤的修复。     

Graft-versus-leukemia effects from donor lymphocyte infusion after nonmyeloablative allogeneic bone marrow transplantation in mice

Background Nonmyeloablative allogeneic bone marrow transplantation has been used since the 1990s as a new hematological stem cell transplantation strategy for treating hematological diseases. The purpose of this study was to explore the graft-versus-leukemia (GVL) effects of donor lymphocyte infusions (DLIs) after nonmyeloablative allogeneic bone marrow transplantations, while assessing the declines in treatment-associated morbidity, mortality, and graft-versus-host disease (GVHD).Methods A total of 615 (H-2k) mice were injected with L615 tumor cells and received 500 cGy (60Coγ-ray) irradiation three days later, followed by an allogeneic bone marrow transplantation (allo-BMT). The allo-grafts consisted of 3×10（7） bone marrow cells and 1×10（7） spleen cells from BALB/C (H-2d) donor mice. Two days after the allo-BMT, the recipient mice were given 200 mg/kg of cyclophosphamide. Subsequently, recipient mice were infused with either donor spleen cells ［2×10（7）］ on day 14 or 21, or donor spleen cells ［5×10（7）］ pretreated with hydrocortisone and cyclosporin A (CsA) in vitro on day 14 post-BMT.Results The median survival time of mice that received DLI on day 21 and pretreated DLI on day 14 post-BMT was longer than that of controls and the day 14 DLI group (P&lt;0.01). No evidence of severe GVHD was observed in the day 21 DLI group nor in the day 14 treated DLI group. Mixed chimerism was confirmed in the day 14 DLI group, the day 14 treated DLI group, and the day 21 DLI group on the thirteenth day post-transplantation; full donor chimerism was observed two weeks after DLI.Conclusion Donor lymphocyte infusion after nonmyeloablative bone marrow transplantation may reduce transplantation-associated morbidity and mortality while strengthening graft-versus-leukemia effects.     

NADH对昆明小鼠辐照后造血系统的影响

目的 探讨NADH对小鼠辐照后造血系统的影响。方法 昆明小鼠60只．随机分为3组，每组20只，实验Ⅰ、Ⅱ和Ⅲ组分别为正常对照组、辐照前注入生理盐水组和辐照前注射NADH药物组。生理盐水和NADH药物均采用腹腔注射，2次／d，照射前3d给药。辐照采用6．0Gy ^60Co一次性辐照小鼠。尾静脉采血计数连续观察辐射后不同时期对外周血白细胞总数的影响；辐照后24h，取股骨骨髓涂片，显微镜下计算骨髓有核细胞数和其分裂指数；通过DNA琼脂糖凝胶电泳观察昆明小鼠辐照后24、48h骨髓细胞凋亡的改变。结果 NADH能够抑制受照后小鼠外周血白细胞减小,增加骨髓有核细胞数及分裂指数,但并不能抑制DNA凋亡片段形成。结论 NADH能够明显抑制辐射诱导的造血系统损伤,但并不通过抑制骨髓细胞凋亡途径。     

rhBMP_2不同动物异位诱导成骨及其小鼠体内诱导成骨的剂量依赖性

目的对大肠杆菌表达的hBMP2在小鼠体内进行剂量依赖性研究,并观察其在大鼠和家兔体内异位诱导成骨情况,为临床应用提供参考。方法大鼠、家兔股部肌袋植入rhBMP21.5和3.0mg,植入后1、2、4周取材观察成骨情况;小鼠股部肌袋植入100~1000μg的rhBMP2,植入后1、2、3周取材,行组织学检查,成骨量分析,碱性磷酸酶和钙含量测定。结果大鼠植入1.5mgrhBMP2植入2周、家兔3.0mgrhBMP2植入4周表现为高效成骨活性。rhBMP2小鼠体内诱导成骨实验表现为成骨量,碱性磷酸酶活性和钙含量显著的剂量依赖性。结论原核表达的hBMP在小鼠、大鼠和家兔体内有高效的诱导成骨活性,小鼠体内实验rhBMP2表现为明显的剂量依赖性。     

rhBMP-2对兔下颌骨放疗后骨延长的影响

目的兔下颌骨放射后骨延长的新生骨形成受到轻微抑制,本研究旨在初步了解rhBMP-2对其促进成骨疗效。方法 50Gy组(对照组)和rhBMP-2组(实验组)各兔6只,均在术前1个月接受50Gy放射,间歇8天后,以0.4毫米/次、2次/天的速度进行牵引,固定期6周。实验组在截骨线内同期置入2.0mg rhBMP-2。结果标本和X线均显示两组成骨均良好,与周围骨质无明显的界限;HE组织学切片可见两组骨小梁稀疏、细小,并存在成片状致密纤维结缔组织。图像分析软件结果是:50Gy组和BMP组的骨小梁面积比分别为0.445±0.054和0.459±0.036。经方差分析,两组间差异无统计学意义。结论本研究中的rhBMP-2不能促进兔下颌骨放射后骨延长的新生骨形成。     

Effects of Ligustrazine on Expression of Bone Marrow Heparan Sulfates in Syngeneic Bone Marrow Transplantation Mice

Hematopoietic reconstitution after bone marrowtransplantation (BMT ) is a complicated process,which concerns notonly with the quality and quanti-ty of hematopoietic stem cells(HSC) ,but also withthe recovery of hematopoietic inducing microenviron-ment (HIM) .The contactof HSC and HIM,whichregulates the proliferation,differentiation and self-maintenance of HSC,is critical for hematopoietic re-constitution.Heparan sulfates (HS) are a kind of gly-cosaminoglycans macromolecules composed of…     

同名异种鸡血藤对60Co γ射线辐射小鼠外周血象的影响

目的:研究密花豆藤(Spatholobus suberectu)、白花油麻藤(Mucuna bird woodiana)、皱果崖豆藤(Millettia oosperma)、香花崖豆藤(Millettia dielsiana)等4种同名异种鸡血藤对骨髓抑制小鼠外周血象的作用。方法:60Coγ射线亚致死量辐照造成骨髓抑制小鼠模型,灌胃给药,于给药后1、3、7、14和21d监测其外周血白细胞、红细胞和血小板计数。结果:给予同名异种鸡血藤治疗后,小鼠外周血白细胞、红细胞和血小板计数下降趋势减缓,与对照组比较,白花油麻藤和密花豆藤高、低剂量组的白细胞、红细胞和血小板计数升高,差异有统计学意义(P<0.05)。结论:同名异种鸡血藤均可在一定程度上促进辐射小鼠外周血象的恢复,其中密花豆藤和白花油麻藤作用相对较好。     

重组人BMP-2骨诱导中神经生长因子的表达

目的通过观察重组人骨形态发生蛋白-2(rhBMP-2)异位诱导成骨过程中神经生长因子(NGF)的表达,探讨NGF在BMP骨诱导中的作用。方法采用ICR小鼠48只,随机分为实验组和对照组,行右侧股后部肌袋异位成骨模型,实验组植入含0．125mg rhBMP-2胶原海绵复合物,对照组仅植入相同体积胶原海绵,分别于术后3、7、14、21d取材,进行放射学、组织学及免疫组化检测。结果放射学和组织学检测均证实,rhBMP-2胶原海绵复合物具有良好的诱导成骨能力;术后第7d,在软骨大量形成期,NGF阳性染色达到高峰．在成纤维细胞、软骨前体细胞、软骨细胞、肥大软骨细胞和成骨细胞中均有阳性表达;术后14、2ld,阳性细胞数量和染色强度有所回落。对照组未见成骨现象。结论首次证实在外源性BMP诱导成骨过程中有明显的NGF表达,提示NGF可能通过直接或间接的方式参与并协同BMP的诱导成骨过程。     

不同成骨诱导作用下骨髓间充质干细胞的基因表达模式

目的研究骨髓间充质干细胞不同成骨分化模式的分子调控机制，为将其作为骨组织缺损基因治疗的载体细胞奠定理论基础。方法用密度梯度离心结合贴壁培养法分离纯化小鼠骨髓间充质干细胞（MSCs），传代扩增后分别用矿化培养基（100nmol／L地塞米松、10mmol／Lβ-甘油磷酸钠和50mg／mL抗坏血酸）与重组人骨形成蛋白-2（rhBMP-2，500ng／mL）进行诱导。分别于诱导后1、2周行茜素红染色鉴定钙结节形成情况；诱导后1、2和3d时用RT-PCR检测Runx2、Osx、OCN和ColⅠ的基因表达情况。结果矿化诱导组和rhBMP-2诱导组，茜素红染色均呈阳性钙结节，但矿化诱导组细胞于1周后形成矿化结节，时间早于rhBMP-2诱导组（2周后形成矿化结节）；RT-PCR显示，在矿化诱导组，Runx2在诱导的1、2、3d均没有表达，Osx在诱导后2、3d有表达，OCN和ColⅠ在诱导的1、2、3d均有表达；在rhBMP-2诱导组．Runx2和Osx于诱导的第2d开始表达，OCN和ColⅠ在诱导的1、2、3d均表达。结论与rhBMP-2相比，矿化培养基通过更为直接的信号传导方式调控着骨髓间充质干细胞向成骨细胞分化，因此矿化培养基诱导骨髓间充质干细胞形成矿化结节的能力强于rhBMP-2。     

The Effects of Ligustrazine on the Expression of bFGF and bFGFR in Bone Marrow in Radiation Injured Mice

Thebonemarrowhematopoiesisinductivemi croenvironmentismadeupofstromalcells ,extracel lularmatrixandmicrangiumsystem .Micrangiumsystemisanimportant partinthebonemarrowhematopoiesisinductivemicroenvironment .Bonemarrowisoneoforganswhicharesensitivetoradia tion .Basicfibroblastgrowthfactor (bFGF)isasin glechain peptide[1] ,whichincludes 15 4aminoacids[2 ] .bFGFbindstoitsspecificreceptor(bFGFR) ,whichstimulatestheproliferationandim migrationofendothelialcells .Itplaysanimportantroleintheembryo…     

Effects of Platelet Factor 4 on Expression of Bone Marrow Heparan Sulfate in Syngenic Bone Marrow Transplantation Mice

Platelet factor 4 ( PF4) is a negativehematopoietic factor.It can inhibit the prolifera-tion of endothelial cells and hematopoietic stem/progenitor cells,particularly megakaryoryocyticcells,reversibly[1] ,inhibit DNA synthesis,blockcell cycle progression during S phase and reducethe sensibility of normal hematopoietic stem/pro-genitor cells,but not some cancer or leukemia celllines,to cytotoxic drugs and ionizing radia-tion[2 - 3] ,and it also can cause a population in-crease of the stem cel…     

Protective effect of an extract of Guipi Pill (归脾丸) against radiation-induced damage in mice

Objective:To study the protective effect of an extract of Guipi Pill(归脾丸) against radiationinduced damage.Methods:A total of 100 Kunming mice were randomly divided into normal group,model group,positive drug group(treated with radioprotective agent "523",5 mg/kg at 24 h before irradiation) and two treatment groups,with 20 mice in each group.The extract of water extraction-alcohol precipitation (WAP) from Guipi Pill were administered orally to the mice in the two treatment groups at the dose of 500 and 1,000 mg/kg,respectively,for 6 days prior to whole body radiation(8 Gy).Fifty mice with 10 in each group were used to observe the survival rate 30 days after radiation.The other 50 mice with 10 in each group were sacrificed on day 10 after radiation(6 Gy) in order to take blood,liver and unilateral femur.Results:Pretreatment prior to irradiation with WAP resulted in a significantly higher 30-day survival rate of mice after exposure to a potentially lethal dose of 8-Gy radiation.WAP could significantly increase the total white blood cell count and DNA content of bone marrow,and it also increased the activity of various antioxidant enzymes,such as superoxide dismutase,catalase,total antioxidant capacity and glutathione peroxidase in liver tissue of mice, which were reduced by radiation treatment.Maleic dialdehyde level and bone marrow micronucleus rate were significantly reduced by WAP,which were increased after 6-Gy radiation.Conclusion:WAP of Guipi Pill could increase the 30-day survival rate and the antioxidant capacity as well as protect bone marrow in mice.WAP of Guipi Pill is an effective radioprotective agent.     

Enhancing Effects of Ligustrazine on Expression of CD31 and Hematopoietic Reconstitution in Syngenic Bone Marrow Transplantation of Mice

Summary： The effect of ligustrazine on the expression of CD31 in syngenic bone marrow transplantation （BMT） mice was studied. Fifty-six Balb/c mice were divided into 3 groups： normal control group. BMT control group, and ligustrazine treated group. Syngenic BMT mouse models were established according to the literatures. In BMT control group and the ligustrazine treated group, the mice were given respecxively orally 0.2 mL saline and 2 mg ligustrazine twice a day. On the 7th, 14th, and 21st day after BMT, the mice were killed. The expression of CD31 on the surface of bone marrow nuclear cells （BMNC） was detected by flow cytometry. Peripheral blood leukocytes, platelets and BMNC were counted. Histological observation of bone marrow was made. The results showed thai in ligustrazine treated group the peripheral blood leukocylcs, platelets and BMNC counts, and the expression of CD31 on the day 7, 14, 21 after BMT were higher than in BMTcontrol group （P〈0.01 or P〈0.05）. In conclusion, ligustrazine could obviously enhance the CD31 expression on the surface of BMNC after syngcnic BMT in mice, which may be one of the mecha- nisms underlying the ligustrazine accelerating hematopoietic reconstitution in syngenic BMT.