钴60-γ射线致小鼠睾丸支持细胞结构及功能的损害

发布时间：2018-04-21 22:34

本文选题：钴60-γ + 支持细胞　；参考：《郑州大学》2013年硕士论文

【摘要】：研究背景及目的：随着工业化进程的加速,日常生活中环境辐射日益加剧,人们对放射性损伤也越来越重视。放射线可损伤全身多个系统,其中性腺组织对放射线极为敏感。支持细胞(Sertoli cells)作为睾丸生精上皮中唯一的非生殖细胞,是维持生精小管内生精环境稳定的重要细胞。因而,任何因素导致睾丸支持细胞结构及功能的损伤都有可能影响精子的形成,甚至导致男性不育。本课题拟通过不同剂量的钴60(60Co)-γ射线局部照射小鼠睾丸,并观察相应指标的变化,以明确不同剂量60Co-γ射线致小鼠睾丸支持细胞结构及功能急性损伤的程度,并初步探讨引起这种损伤的机制。方法：48只健康雄性KM小鼠,采用随机数字表法平均分为4组(n=12)：正常对照组(A组)；60Co-γ射线双侧睾丸局部照射组有3个剂量组：剂量2Gy(B组)；剂量6Gy(C组)；剂量10Gy(D组),局部均匀照射下腹部,时间设定为517秒。照射完成48h后,处死小鼠,测定实验小鼠身体及双睾丸重量并采集血液3ml；采用酶联免疫吸附法(ELISA)检测血清中生殖激素,即：促黄体生成素(luteinizing hormone, LH)、卵泡刺激素(follicle-stimulating hormone, FSH)以及抑制素β (Inhibin0)水平；应用光学显微镜观察小鼠睾丸组织形态学变化；应用实时荧光定量PCR法(Real-time PCR)检测实验小鼠睾丸组织中抑制素β mRNA的相对表达量；采用酶联免疫吸附法(ELISA)检测实验小鼠睾丸组织中抗氧化指标,即：还原性谷胱甘肽(Glutathione, GSH)、超氧化物歧化酶(Superoxide dismutase, SOD)、丙二醛(Malondialdehyde, MDA)含量。结果：60Co-γ射线照射组(B、C、D组)较正常组(A组)睾丸重量下降(P0.05),以C、D两组更为明显(P0.01)；血清生殖激素FSH水平明显上升(P0.05),而抑制素p水平明显下降(P0.05)；光镜下可见小鼠睾丸生精细胞及支持细胞均发生不同程度的损伤,其中以D组损伤程度最为明显；睾丸组织抑制素βmRNA表达水平明显下降,差异均有统计学意义(allP0.05);睾丸组织SOD含量呈下降趋势(P0.05),其中C、D两组下降较为明显(allP0.05);GSH含量亦呈下降趋势,差异均有统计学意义(allP0.05), MDA含量呈上升趋势,差异均有统计学意义(allP0.05)。结论：60Co-γ射线对睾丸支持细胞结构及功能有明显损伤作用,且损伤程度与照射剂量呈正相关性；损伤的机制与射线致睾丸抗氧化系统受损,引起睾丸组织氧化-抗氧化系统失衡导致氧化应激损伤作用有关。
[Abstract]:Background and purpose: with the acceleration of industrialization, the environmental radiation in daily life is getting more and more serious, and people pay more and more attention to the radiation damage. Radiation can damage multiple systems throughout the body, in which gonadal tissue is extremely sensitive to radiation. Sertoli cells, as the only non-germ cells in spermatogenic epithelium of testis, are important cells to maintain stable spermatogenic environment in seminiferous tubules. Therefore, any cause of testicular Sertoli cell structure and function damage may affect the formation of sperm, and even lead to male infertility. In order to determine the degree of acute injury of testicular Sertoli cell structure and function induced by different doses of 60Co- 纬 ray, the mice testis were irradiated locally with different doses of cobalt 60Co- 纬 ray and the changes of corresponding indexes were observed. The mechanism of the injury was also discussed. Methods A total of 48 healthy male km mice were used. The method of random digital table was used to divide into 4 groups on average: normal control group, group A, group A, group A: dose 2Gy(B group, dose group 6Gy(C group, dose group 10Gy(D group, local uniform irradiation of lower abdomen, time was set at 517 seconds. After 48 hours of irradiation, the mice were killed, the weight of the body and testis of the experimental mice were measured, and the blood was collected 3 ml. The serum reproductive hormones were detected by Elisa. The levels of luteinizing hormone, luteinizing hormone, follicle-stimulating hormone, follicle-stimulating hormone and inhibin 尾 -Inhibin0 were observed by optical microscope. Real-time PCR method was used to detect the relative expression of inhibin 尾 mRNA in testis of experimental mice, and enzyme linked immunosorbent assay (Elisa) was used to detect the antioxidant index in testis of experimental mice. The contents of Glutathione, GSH, Superoxide dismutase, SODX, malondialdehyde (MDA) and malondialdehyde (MDAs) were observed in Glutathione, glutathione, superoxide dismutase (SOD), malondialdehyde (MDA) and malondialdehyde (MDAs). Results compared with the normal group, the weight of testis was decreased in the weight of testis, especially in the two groups. The level of serum reproductive hormone (FSH) increased significantly (P0.05), while the level of inhibin P decreased significantly (P0.05). Under light microscope, the testis of mice were found to be testicular. Spermatocytes and Sertoli cells were damaged to varying degrees. In group D, the degree of injury was the most obvious, the expression of inhibin 尾 mRNA in testis was obviously decreased, the difference was statistically significant, and the content of SOD in testis showed a decreasing trend (P 0.05). The difference was statistically significant (P 0.05), and the MDA content was on the rise, and the difference was statistically significant (P 0.05). Conclusion the structure and function of testicular Sertoli cells were significantly damaged by W60Co- 纬 rays, and the degree of injury was positively correlated with the dose of irradiation, and the mechanism of injury was related to the damage of antioxidant system of testis induced by radiation. It is related to the oxidative stress damage caused by the imbalance of oxidation-oxidation system in testis.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R818.74

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