小鼠骨髓造血相关因子-sIL1rn和GPx7基因的克隆与功能研究
发布时间：2018-12-19 11:08
【摘要】：白细胞介素超家族和谷胱甘肽过氧化物酶是重要的细胞因子，在研究中我们发现白介素1受体拮抗剂(sIL1rn)和谷胱甘肽过氧化物酶(Gpx7)在造血干细胞(HSCs)扩增中具有正调节因子的作用，能够有效促进HSCs的扩增。 首先我们通过注射5—氟尿嘧啶(5-FU)建立了小鼠造血应急反应模型，展示了骨髓抑制和再生过程。并运用基因芯片分析方法，，对此模型中第0、3、7、11、14天的基因表达差异进行比较研究，获得了小鼠骨髓抑制和再生的基因表达图谱。根据这个图谱，我们发现了一组在此过程中表现出与造血调节功能明显相关的细胞因子基因。本文对其中的两个基因—sIL1rn和Gpx7基因进行了克隆。将sIL1rn接入真核表达载体phCMV，同时将Gpx7接入真核表达载体pcDNA3.1，并通过肌肉注射、电击穿孔、体内表达等方法获得了该基因在小鼠胫前肌的高效表达，初步研究了sIL1rn和Gpx7在骨髓造血过程中的功能。结果表明，体内表达sIL1rn能显著增加5-FU注射后小鼠外周血白细胞数，能将骨髓抑制时间由14天缩短到11天左右。Gpx7可以明显增加骨髓细胞恢复数量。同时，对基因芯片发现的与sIL1rn具有一定协同作用的生长激素，利用皮下注射融合蛋白的方法，也进行了初步的功能研究。
[Abstract]:Interleukin-superfamily and glutathione peroxidase are important cytokines. We found that interleukin-1 receptor antagonists (sIL1rn) and glutathione peroxidase (Gpx7) play a positive role in the (HSCs) amplification of hematopoietic stem cells and can effectively promote the expansion of HSCs. First, we established a mouse hematopoietic emergency response model by injecting 5-fluorouracil (5-FU) to demonstrate the process of bone marrow suppression and regeneration. Using the method of gene chip analysis, the difference of gene expression in the model was studied on day 71114, and the gene expression map of bone marrow suppression and regeneration in mice was obtained. Based on this map, we found a set of cytokine genes that were significantly associated with hematopoietic regulation. In this paper, two genes, sIL1rn and Gpx7, were cloned. SIL1rn was inserted into the eukaryotic expression vector phCMV, and Gpx7 was inserted into the eukaryotic expression vector pcDNA3.1,. The gene was expressed in the anterior tibial muscle of mice by intramuscular injection, electroporation and in vivo expression. The function of sIL1rn and Gpx7 in bone marrow hematopoiesis was studied. The results showed that the expression of sIL1rn in vivo could significantly increase the number of leukocytes in peripheral blood of mice injected with 5-FU and shorten the time of bone marrow suppression from 14 days to about 11 days. Gpx7 could significantly increase the number of bone marrow cells recovered. At the same time, the function of growth hormone with sIL1rn was studied by subcutaneous injection of fusion protein.
【学位授予单位】：沈阳药科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R346
本文编号：2386841
[Abstract]:Interleukin-superfamily and glutathione peroxidase are important cytokines. We found that interleukin-1 receptor antagonists (sIL1rn) and glutathione peroxidase (Gpx7) play a positive role in the (HSCs) amplification of hematopoietic stem cells and can effectively promote the expansion of HSCs. First, we established a mouse hematopoietic emergency response model by injecting 5-fluorouracil (5-FU) to demonstrate the process of bone marrow suppression and regeneration. Using the method of gene chip analysis, the difference of gene expression in the model was studied on day 71114, and the gene expression map of bone marrow suppression and regeneration in mice was obtained. Based on this map, we found a set of cytokine genes that were significantly associated with hematopoietic regulation. In this paper, two genes, sIL1rn and Gpx7, were cloned. SIL1rn was inserted into the eukaryotic expression vector phCMV, and Gpx7 was inserted into the eukaryotic expression vector pcDNA3.1,. The gene was expressed in the anterior tibial muscle of mice by intramuscular injection, electroporation and in vivo expression. The function of sIL1rn and Gpx7 in bone marrow hematopoiesis was studied. The results showed that the expression of sIL1rn in vivo could significantly increase the number of leukocytes in peripheral blood of mice injected with 5-FU and shorten the time of bone marrow suppression from 14 days to about 11 days. Gpx7 could significantly increase the number of bone marrow cells recovered. At the same time, the function of growth hormone with sIL1rn was studied by subcutaneous injection of fusion protein.
【学位授予单位】：沈阳药科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R346
【参考文献】
相关期刊论文 前1条
1 郭甫坤,吴曙光;白介素1信号转导研究进展[J];国外医学.分子生物学分册;2001年01期
本文编号：2386841
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