X射线照射对子宫内膜胚胎黏附功能的影响

发布时间：2018-03-06 09:53

本文选题：X射线　切入点：子宫内膜　出处：《大连医科大学》2012年硕士论文　论文类型：学位论文

【摘要】：目的： X射线是人类日常生活中的常见射线。X射线也是一种电离辐射形式。研究表明：X射线的穿透性强，因此可导致受照射组织的病理损伤和功能障碍。研究发现，X射线照射可使在早期形成期的胚胎出现发育阻滞、畸形、植入失败或胚胎死亡，特别是处于生长发育的局部器官。除此之外，还可引起白血病等疾病的发生。近年来，，不孕症的发生率不断上升，约占育龄妇女的10-15%[1]。尽管推测与使用和接触具有电离辐射的电子产品等有关，但尚未完全证实。因此，研究X射线对子宫内膜胚胎接受性的影响有一定必要性。本实验拟利用X射线照射子宫内膜细胞，通过对p53及其下游靶蛋白的检测，及利用体外胚胎黏附模型分析X射线对胚胎细胞黏附功能的影响，探寻X射线影响胚胎细胞间黏附的分子机制。方法：（1）培养接受态人子宫内膜上皮细胞RL95-2，利用单细胞凝胶电泳（SCGE）实验，检测不同剂量（1Gy，10Gy）X射线照射后RL95-2细胞的DNA链断裂情况；（2）Western Blot检测细胞不同剂量照射后提取蛋白中的p53及其下游信号通路蛋白的表达情况；（3）培养并制备RL95-2单层细胞，并用不同剂量X射线处理，分别加入胚胎细胞（JAR），构成体外胚胎黏附模型，分析胚胎细胞与不同处理组的子宫内膜间的黏附率。结果：（1）单细胞凝胶电泳实验结果显示，在经过不同剂量X射线处理后，RL95-2细胞出现拖尾现象，且高剂量组彗星尾长、尾距以及尾DNA%较对照组具有显著性差异，RL95-2细胞内DNA链断裂程度随X射线照射剂量升高而增加；（2）Western Blot结果显示，经X射线处理后的细胞中p53蛋白明显增多，高剂量组蛋白量较低剂量组积累情况较为明显，而下游蛋白p21、p27基本无变化；（3）JAR细胞与高剂量X射线处理RL95-2细胞黏附率明显低于低剂量组，而空白对照组则高于处理组。结论： X射线照射可使RL95-2细胞内的DNA链发生断裂，并使p53蛋白表达增加，而对p21、p27蛋白表达的影响不明显。X射线照射后降低对子宫内膜细胞与胚胎细胞间的黏附功能。研究提示：X射线照射引起的DNA断裂性损伤是导致子宫内膜胚胎接受性降低的重要原因之一。本研究为探讨X射线照射与不孕症的相关性提供实验依据。
[Abstract]:Objective:. X-rays are a common form of ionizing radiation in our daily lives. This may lead to pathological damage and dysfunction in irradiated tissues. Studies have found that X-ray exposure can cause developmental arrest, malformation, implantation failure, or embryo death in the early stages of formation. Especially in the local organs of growth and development. In addition, it can also cause the occurrence of diseases such as leukemia. In recent years, the incidence of infertility has been increasing. About 10-15% of women of childbearing age [1]. Although it is supposed to be related to the use and exposure of electronic products with ionizing radiation, it has not been fully confirmed. Therefore, it is necessary to study the effect of X-ray on the acceptability of endometrial embryos. The aim of this study was to study the effects of X ray irradiation on the adhesion function of endometrial cells by detecting p53 and its downstream target proteins and using an in vitro embryo adhesion model. To explore the molecular mechanism of X-ray effect on intercellular adhesion of embryos. Methods:. Human endometrial epithelial cells (RL95-2) were cultured by single cell gel electrophoresis (SCGE). The DNA strand breakage of RL95-2 cells irradiated with different doses of 1 Gyr1 Gyr 10 Gy X ray was detected. Western Blot was used to detect the expression of p53 and its downstream signaling pathway proteins in the extracted proteins after different doses of irradiation. RL95-2 monolayers were cultured and prepared. Different doses of X-rays were added to JARA cells to form an in vitro embryo adhesion model, and the adhesion rate between embryonic cells and endometrium of different treatment groups was analyzed. Results:. 1) the results of single cell gel electrophoresis (SCGE) showed that the tail of RL95-2 cells was delayed after different doses of X-ray, and the tail length of comets in the high dose group was longer than that in the control group. The DNA strand breaking degree of RL95-2 cells increased with the increase of the dose of X-ray irradiation. The results of Western Blot showed that the p53 protein in RL95-2 cells treated with X-ray was significantly increased. The accumulation rate of high dose histone was significantly higher than that of low dose group, while the adhesion rate of downstream protein p21 P27 to high dose X ray treated RL95-2 cells was significantly lower than that of low dose group, while that of blank control group was higher than that of control group. Conclusion:. X-ray irradiation could break the DNA chain and increase the expression of p53 protein in RL95-2 cells. However, there was no significant effect on the expression of p21, p27 protein. The adhesion function between endometrial cells and embryonic cells was decreased after X-ray irradiation. It was suggested that the damage of DNA fragmentation induced by W ~ (-1) X-ray irradiation might lead to the receptivity of endometrial embryos. This study provides an experimental basis for the study of the correlation between X-ray irradiation and infertility.
【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R818

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