| 陈婷婷,张西志,汪步海,花威,杨占山.耐辐射球菌pprI基因对γ射线损伤小鼠的抗辐射机制[J].中华放射医学与防护杂志,2013,33(2):119-123 |
| 耐辐射球菌pprI基因对γ射线损伤小鼠的抗辐射机制 |
| The mechanism of Deinococcus radiodurans pprI gene in enhancing mice radioresistance to γ-rays |
| 投稿时间：2012-09-25 |
| DOI：10.3760/cma.j.issn.0254-5098.2013.02.002 |
| 中文关键词: 耐辐射球菌 pprI基因 转基因 放射损伤 |
| 英文关键词:Deinococcus radiodurans pprI gene in vivo electroporation Radiation injury |
| 基金项目:国家自然科学基金(81072236);国防基础科研资助项目(A3820060138);江苏高校优势学科建设工程资助项目 |
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| 中文摘要: |
| 目的 研究耐辐射球菌pprI基因活体电转染在哺乳动物γ射线放射损伤中发挥抗辐射作用的机制。方法 采用雄性昆明种小鼠,随机区组法分为健康对照组、单纯照射组、空载体转染组和基因转染组。将自行构建的含有pprI基因的pCMV-HA-pprI质粒注入接受6 Gy γ射线照射的小鼠肌肉内,然后采用活体基因电转导技术将该基因导入细胞内,应用Western blot法对PprI蛋白及其下游基因recA在哺乳动物细胞中的同源类似物Rad51以及Rad52蛋白表达进行检测。结果 照后1 d基因转染组小鼠肌肉组织中 PprI蛋白明显表达,7 d后未见蛋白表达,其余组小鼠未见蛋白表达;在基因转染组小鼠,肺脏Rad51蛋白于照后第1、7和14天明显表达,明显高于单纯照射组及空载体转染组小鼠,肝脏Rad51蛋白于照后第1天和第28天明显表达,肾脏Rad51蛋白于照后第1天及第14天明显表达;在各组小鼠的肺脏、肝脏组织中检测了Rad52蛋白,其表达无明显规律。结论 耐辐射球菌pprI原核基因成功地在哺乳动物体内细胞表达了PprI蛋白,并作用于下游基因recA在哺乳动物细胞中的同源类似物rad51基因,使其蛋白表达显著增高而发挥其抗哺乳动物辐射损伤的作用。 |
| 英文摘要: |
| Objective To investigate the mechanism of Deinococcus radiodurans pprI gene in enhancing mice radioresistance to γ-rays by transfecting it in vivo.Methods The male Kunming mice were randomly divided into control group, irradiated group, pCMV-HA transfected group and pCMV-HA-pprI transfected group. The pCMV-HA-pprI plasmid contained pprI gene was injected into the muscle of mice which were exposed to total 6 Gy of γ-ray irradiation. After injection, the in vivo gene electroporation technology was used to transfect the pprI gene into the cells, and Western blot was used to identify the PprI protein, mammalian homolog protein Rad51 corresponding to recA gene downstream of pprI, and protein Rad52.Results In the muscle of the mice of transfected pCMV-HA-pprI group, the protein PprI expressed significantly at 1 d post-irradiation, but there was no expression of pprI gene 7 d post-irradiation and in other groups. In the mice of transfected with pCMV-HA-pprI, the expression of Rad51 protein was significantly increased in the lungs at 1, 7 and 14 d post-irradiation, and significantly increased in the liver at 1 and 28 d post-irradiation and increased in the kidneys at 1 and 14 d post-irradition. However, there was no obvious change of Rad52 protein expression in the lungs and livers of mice in all groups.Conlusions The prokaryotic gene pprI could act on the mammalian homologisation analogues rad51 gene downstream of recA gene and then increase the expression level of protein Rad51 which results in the enhancement of radioresistance. |
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