抗辐射球菌pprI基因活体电转染对受照小鼠睾丸病理学作用的研究  

Pathological study on the testis of mice irradiated by γ-rays after transfecting pprI gene by in vivo electroporation

作　　者：连利霞[1] 陈婷婷[1] 张永芹[1] 王秀珍[1] 杨占山[1] 

机构地区：[1]苏州大学医学部放射医学与公共卫生学院放射毒理教研室,苏州215123

出　　处：《辐射研究与辐射工艺学报》2011年第1期17-22,共6页Journal of Radiation Research and Radiation Processing

基　　金：国防基础科研项目(A3820060138);国家自然科学基金项目(30570549)资助

摘　　要：观察抗辐射球菌pprI基因活体电转染小鼠受γ射线辐照后睾丸组织的形态学变化,探讨一种新的原核基因表达对哺乳动物生殖系统损伤的防治作用。将含有pprI基因的pCMV-HA-pprI质粒注入小鼠肌肉内,然后采用活体基因电转染技术将该基因导入细胞内,对照组小鼠和转基因组小鼠均采用60Coγ射线全身照射,吸收剂量为6 Gy,分别于辐照后第1、7、14、28和35天取下注射部位肌肉组织,提取组织总蛋白,进行PprI蛋白的Western blotting检测。同时取小鼠睾丸组织,石蜡包埋制片,苏木精-伊红染色,进行镜下病理学观察。PprI蛋白在辐照后第1天有较强的表达,说明pprI质粒成功转染至小鼠体内。对照组小鼠在照后第1天生精小管结构完整,各级生精细胞未见明显病理变化;第7天在部分生精小管观察到少量精原细胞坏死;第14天生精细胞变性坏死明显加重,生精小管萎缩变细,相互"远离";至第28天各级生精细胞及精子极度减少,生精小管内细胞分层结构基本消失,甚至空虚,仅残留支持细胞,间质疏松;第35天空虚的生精小管内开始出现新生精原细胞。而转基因组小鼠辐照后第1天生精细胞未见明显病理变化;第7天部分生精小管见少量精原细胞坏死,较单纯照射组轻;第14天生精细胞变性坏死加重,各级生精细胞数量减少;第28天局部生精小管内出现新生精原细胞,基底层结构基本显现;第35天转基因组局部生精小管内新生生精细胞增多,出现分层现象。研究结果表明,pprI基因活体电转染对小鼠睾丸γ辐射损伤具有显著的防治作用,睾丸辐射损伤修复明显提前。To investigate the effects of pprl gene from Deinococcus radiodurans transferred by in vivo electroporation on γ-ray injury of mice, the morphological changes of testis in the mice were observed. The pCMV-HA-pprI plasmid containing pprI gene was injected into the muscle of mice. The pprl gene was transfected into the cells by in vivo gene electroporation technology. Then the control group and the transferred pCMV-HA- pprI group were exposed to γ-ray radiation of 6 Gy. The muscle tissue at the site of the injection and the testis tissue were taken on days 1, 7, 14, 28 and 35 after radiation, Then total protein was extracted and used to test the expression of PprI with western blotting technology. The testis specimen prepared by haematoxylin-eosin staining was then examined by light microscopy. The expression of Pprl is remarkable on the 1st day after irradiation to prove that the pprI gene was successfully transfected into the mice. On the 1st day after irradiation there was no obvious pathological change of the testis tissue of the control group. On the 7th day there was degeneration and necrosis of some spermatogonia and spermatocytes in sections of tubules. On the 14th day, the reduction of spermatogonia was generally marked, and there was considerable reduction in the number of primary spermatocytes associated with atrophy of the seminiferous tubules. On the 28th day there was complete depletion of spermatogenic epithelium when spermatocytes and spermatids had largely disappeared, with no regeneration of spermatogonia and only sertoli cells nuclei remaining along the basement membrane. On the 35th day, spermatogonia were actively regenerating in some of the tubules. Compared with the control group, there was also no significant difference on the 1st after irradiation in the transgenic animal. On the 7th day the degeneration and necrosis of some spermatogonia and spermatocytes in sections of tubules was less than that of the control group. On the 14th day the reduction of spermatogonia was generally marked, a

关 键 词：抗辐射球菌 PPRI 活体电穿孔 睾丸组织 病理学 

分 类 号：R818.021[医药卫生—放射医学] R818.059[医药卫生—临床医学]