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出血是大剂量电离辐射后所致损伤的主要临床症状之一 ,且可发展成致命的出血综合征。国内外均尝试在辐射或化疗动物模型上用一些细胞因子促进血小板恢复 ,以减少出血。但由于多次注射等不尽人意 ,因此人们仍在进行新措施的研究。用病毒载体进行基因治疗有一定的优势 ,但也有可能增加感染的几率。本实验尝试运用质粒载体对辐射损伤进行基因治疗。本研究观察了辐射对体内质粒载体基因转移效率的影响 ,以及转移hIL 6基因后对照射小鼠造血恢复的影响。经 6 .5Gy照射的小鼠一次肌内注射表达IL 6的质粒DNA后 ,用ELISA方法检测血浆IL 6水平 ,以蛋白表达量评价基因转移效率。结果显示 ,治疗组从照后 4天hIL 6开始增加 ,于照后 11天左右达峰值 ,而此时对照组仍处于低水平。治疗组照后 2 8天时蛋白表达仍维持在较高水平。不同照射剂量治疗组的结果相比 ,7.5Gy照射组hIL 6水平约为 5 .0Gy照射组的两倍。体内高表达的hIL 6明显促进照射小鼠造血恢复 ,治疗组不仅血小板的最低值明显提高 ,而且骨髓细胞中CFU GEMM和CFU Meg也明显增加。外周血血小板数的增加部分是由于网织血小板的增加 ,说明巨核细胞释放血小板的速度加快。以上结果表明电离辐射可以明显提高IL 6质粒DNA的体内转染效率 ,为质粒载体基因的方法治疗辐射?
Bleeding is one of the major clinical symptoms of injury after a large dose of ionizing radiation and can develop into a fatal bleeding syndrome. At home and abroad are trying to use radiation or chemotherapy in animal models with some cytokines to promote platelet recovery in order to reduce bleeding. However, due to unsatisfactory results such as multiple injections, research on new measures is still under way. Gene therapy with a viral vector has some advantages, but it is also possible to increase the chance of infection. This experiment attempts to use plasmid vector for gene therapy of radiation injury. This study investigated the effects of radiation on plasmid vector gene transfer efficiency in vivo and the effect of hIL 6 gene transfer on hematopoiesis in irradiated mice. The 6 .5Gy irradiated mice were intramuscularly injected IL6 plasmid DNA, the plasma IL-6 levels were measured by ELISA, and the gene transfer efficiency was evaluated by the protein expression level. The results showed that the treatment group from 4 days after the start of hIL 6 increased in about 11 days after irradiation reached its peak, while the control group is still low. In the treatment group, the protein expression maintained at a high level on the 28th day after irradiation. Compared with the results of different irradiation dose treatment group, the hIL 6 level in the 7.5 Gy irradiation group was about twice of that in the 5 Gy irradiation group. In vivo high expression of hIL 6 significantly promote hematopoietic recovery in irradiated mice, the treatment group not only significantly increased the minimum platelet, and bone marrow CFU GEMM and CFU Meg also significantly increased. Part of the increase in peripheral blood platelets is due to an increase in reticulated platelets, indicating that the release of platelets by megakaryocytes is accelerated. The above results show that ionizing radiation can significantly improve the in vivo transfection efficiency of IL 6 plasmid DNA and treat the radiation as a plasmid vector gene method.