促血管生成素-1与促血管生成素-2对造血干细胞动员效应对比研究
发布时间：2019-06-05 14:09
【摘要】：目的比较血管生成素-1与血管生成素-2对造血干细胞动员效率。方法本研究通过构建过表达Ang-2因子的重组腺病毒,使用C57BL/6小鼠作为研究对象,分为空白对照组,阴性病毒对照组、Ang-1组、Ang-2组,通过尾静脉分别注射PBS、对照阴性病毒、Ang-1重组腺病毒、Ang-2重组腺病毒,在第14天收集外周血及骨髓,采用酶联免疫吸附试验分析外周血血清Ang-1浓度及Ang-2浓度,血细胞计数分析外周血白细胞数目变化,流式细胞术分析外周血及骨髓Lin-Sca-1+c-Kit+(LSK)细胞比例,造血祖细胞培养比较外周血集落形成单位的变化,从而比较Ang-1与Ang-2影响造血干细胞动员的效率。结果:1.包装获得的重组Ang-2腺病毒滴度为1×1011pfu/ml,重组腺病毒转染HEK293细胞后可以正常表达荧光基因GFP,分泌Ang-2因子。2.注射重组Ang-1腺病毒及重组Ang-2腺病毒,外周血及骨髓内Ang-1浓度及Ang-2浓度均高于对照组。3.腺病毒组外周血白细胞数目均比空白对照组有大幅度升高(P㩳0.05),而Ang-2组与Ang-1组比阴性病毒组高。4.流式细胞术分析LSK细胞,Ang-1组与Ang-2组外周血LSK细胞均比对照组有大幅度提高(P㩳0.05),Ang-1组比Ang-2组稍高(P㧐0.05);骨髓内Ang-1组与Ang-2组LSK细胞均比对照组增高明显(P㩳0.05),Ang-2组比Ang-1组高(P㩳0.05)。5.外周血造血祖细胞集落分析提示空白对照组与阴性病毒对照组集落单位无差别;Ang-1组与Ang-2组均高于对照组(P㩳0.05),而Ang-1组与Ang-2组无差别。结论Ang-1及Ang-2因子浓度的升高均可以影响造血干细胞动员效率,而Ang-1主要促进造血干细胞动员进入外周血循环,而Ang-2主要促进骨髓内造血干细胞的扩增,Ang-2动员效率比Ang-1稍低。
[Abstract]:Objective to compare the mobilization efficiency of angiogenin-1 and angiogenin-2 on hematopoietic stem cells. Methods in this study, recombinant adenoviruses overexpressing Ang-2 factor were constructed and C57BL/6 mice were divided into blank control group, negative virus control group, Ang-1 group and Ang-2 group. PBS, was injected into tail vein respectively. Peripheral blood and bone marrow were collected on the 14th day compared with negative virus, Ang-1 recombinant adenovirus and Ang-2 recombinant adenovirus. the serum Ang-1 and Ang-2 concentrations in peripheral blood were analyzed by enzyme-linked immunosorbent assay (Elisa). The changes of leukocyte number in peripheral blood were analyzed by blood cell count, the proportion of Lin-Sca-1 c-Kit (LSK) cells in peripheral blood and bone marrow was analyzed by flow cytometry, and the changes of peripheral blood colony forming unit were compared in hematopoietic progenitor cell culture. The effects of Ang-1 and Ang-2 on the mobilization of hematopoietic stem cells were compared. Result: 1. The titer of recombinant Ang- 2 adenovirus obtained by packaging was 1 脳 1011pfuml. The recombinant adenoviruses could normally express fluorescent gene GFP, and secrete Ang-2 factor in HEK293 cells. 2. The concentrations of Ang-1 and Ang-2 in peripheral blood and bone marrow were higher than those in the control group. 3. The concentration of Ang-1 and Ang-2 in peripheral blood and bone marrow were higher than those in the control group. The number of leukocytes in peripheral blood of adenoviral group was significantly higher than that of blank control group (P 鈮，
本文编号：2493585
[Abstract]:Objective to compare the mobilization efficiency of angiogenin-1 and angiogenin-2 on hematopoietic stem cells. Methods in this study, recombinant adenoviruses overexpressing Ang-2 factor were constructed and C57BL/6 mice were divided into blank control group, negative virus control group, Ang-1 group and Ang-2 group. PBS, was injected into tail vein respectively. Peripheral blood and bone marrow were collected on the 14th day compared with negative virus, Ang-1 recombinant adenovirus and Ang-2 recombinant adenovirus. the serum Ang-1 and Ang-2 concentrations in peripheral blood were analyzed by enzyme-linked immunosorbent assay (Elisa). The changes of leukocyte number in peripheral blood were analyzed by blood cell count, the proportion of Lin-Sca-1 c-Kit (LSK) cells in peripheral blood and bone marrow was analyzed by flow cytometry, and the changes of peripheral blood colony forming unit were compared in hematopoietic progenitor cell culture. The effects of Ang-1 and Ang-2 on the mobilization of hematopoietic stem cells were compared. Result: 1. The titer of recombinant Ang- 2 adenovirus obtained by packaging was 1 脳 1011pfuml. The recombinant adenoviruses could normally express fluorescent gene GFP, and secrete Ang-2 factor in HEK293 cells. 2. The concentrations of Ang-1 and Ang-2 in peripheral blood and bone marrow were higher than those in the control group. 3. The concentration of Ang-1 and Ang-2 in peripheral blood and bone marrow were higher than those in the control group. The number of leukocytes in peripheral blood of adenoviral group was significantly higher than that of blank control group (P 鈮，
本文编号：2493585
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