【摘要】：目的比較血管生成素-1與血管生成素-2對造血干細(xì)胞動員效率。方法本研究通過構(gòu)建過表達(dá)Ang-2因子的重組腺病毒,使用C57BL/6小鼠作為研究對象,分為空白對照組,陰性病毒對照組、Ang-1組、Ang-2組,通過尾靜脈分別注射PBS、對照陰性病毒、Ang-1重組腺病毒、Ang-2重組腺病毒,在第14天收集外周血及骨髓,采用酶聯(lián)免疫吸附試驗分析外周血血清Ang-1濃度及Ang-2濃度,血細(xì)胞計數(shù)分析外周血白細(xì)胞數(shù)目變化,流式細(xì)胞術(shù)分析外周血及骨髓Lin-Sca-1+c-Kit+(LSK)細(xì)胞比例,造血祖細(xì)胞培養(yǎng)比較外周血集落形成單位的變化,從而比較Ang-1與Ang-2影響造血干細(xì)胞動員的效率。結(jié)果:1.包裝獲得的重組Ang-2腺病毒滴度為1×1011pfu/ml,重組腺病毒轉(zhuǎn)染HEK293細(xì)胞后可以正常表達(dá)熒光基因GFP,分泌Ang-2因子。2.注射重組Ang-1腺病毒及重組Ang-2腺病毒,外周血及骨髓內(nèi)Ang-1濃度及Ang-2濃度均高于對照組。3.腺病毒組外周血白細(xì)胞數(shù)目均比空白對照組有大幅度升高(P㩳0.05),而Ang-2組與Ang-1組比陰性病毒組高。4.流式細(xì)胞術(shù)分析LSK細(xì)胞,Ang-1組與Ang-2組外周血LSK細(xì)胞均比對照組有大幅度提高(P㩳0.05),Ang-1組比Ang-2組稍高(P㧐0.05);骨髓內(nèi)Ang-1組與Ang-2組LSK細(xì)胞均比對照組增高明顯(P㩳0.05),Ang-2組比Ang-1組高(P㩳0.05)。5.外周血造血祖細(xì)胞集落分析提示空白對照組與陰性病毒對照組集落單位無差別;Ang-1組與Ang-2組均高于對照組(P㩳0.05),而Ang-1組與Ang-2組無差別。結(jié)論Ang-1及Ang-2因子濃度的升高均可以影響造血干細(xì)胞動員效率,而Ang-1主要促進(jìn)造血干細(xì)胞動員進(jìn)入外周血循環(huán),而Ang-2主要促進(jìn)骨髓內(nèi)造血干細(xì)胞的擴(kuò)增,Ang-2動員效率比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，

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