本文選題：X射線　切入點(diǎn)：子宮內(nèi)膜　出處：《大連醫(yī)科大學(xué)》2012年碩士論文　論文類型：學(xué)位論文

【摘要】：目的： X射線是人類日常生活中的常見射線。X射線也是一種電離輻射形式。研究表明：X射線的穿透性強(qiáng)，因此可導(dǎo)致受照射組織的病理損傷和功能障礙。研究發(fā)現(xiàn)，X射線照射可使在早期形成期的胚胎出現(xiàn)發(fā)育阻滯、畸形、植入失敗或胚胎死亡，特別是處于生長發(fā)育的局部器官。除此之外，還可引起白血病等疾病的發(fā)生。近年來，，不孕癥的發(fā)生率不斷上升，約占育齡婦女的10-15%[1]。盡管推測與使用和接觸具有電離輻射的電子產(chǎn)品等有關(guān)，但尚未完全證實(shí)。因此，研究X射線對子宮內(nèi)膜胚胎接受性的影響有一定必要性。 本實(shí)驗(yàn)擬利用X射線照射子宮內(nèi)膜細(xì)胞，通過對p53及其下游靶蛋白的檢測，及利用體外胚胎黏附模型分析X射線對胚胎細(xì)胞黏附功能的影響，探尋X射線影響胚胎細(xì)胞間黏附的分子機(jī)制。 方法： （1）培養(yǎng)接受態(tài)人子宮內(nèi)膜上皮細(xì)胞RL95-2，利用單細(xì)胞凝膠電泳（SCGE）實(shí)驗(yàn)，檢測不同劑量（1Gy，10Gy）X射線照射后RL95-2細(xì)胞的DNA鏈斷裂情況；（2）Western Blot檢測細(xì)胞不同劑量照射后提取蛋白中的p53及其下游信號通路蛋白的表達(dá)情況；（3）培養(yǎng)并制備RL95-2單層細(xì)胞，并用不同劑量X射線處理，分別加入胚胎細(xì)胞（JAR），構(gòu)成體外胚胎黏附模型，分析胚胎細(xì)胞與不同處理組的子宮內(nèi)膜間的黏附率。 結(jié)果： （1）單細(xì)胞凝膠電泳實(shí)驗(yàn)結(jié)果顯示，在經(jīng)過不同劑量X射線處理后，RL95-2細(xì)胞出現(xiàn)拖尾現(xiàn)象，且高劑量組彗星尾長、尾距以及尾DNA%較對照組具有顯著性差異，RL95-2細(xì)胞內(nèi)DNA鏈斷裂程度隨X射線照射劑量升高而增加；（2）Western Blot結(jié)果顯示，經(jīng)X射線處理后的細(xì)胞中p53蛋白明顯增多，高劑量組蛋白量較低劑量組積累情況較為明顯，而下游蛋白p21、p27基本無變化；（3）JAR細(xì)胞與高劑量X射線處理RL95-2細(xì)胞黏附率明顯低于低劑量組，而空白對照組則高于處理組。 結(jié)論： X射線照射可使RL95-2細(xì)胞內(nèi)的DNA鏈發(fā)生斷裂，并使p53蛋白表達(dá)增加，而對p21、p27蛋白表達(dá)的影響不明顯。X射線照射后降低對子宮內(nèi)膜細(xì)胞與胚胎細(xì)胞間的黏附功能。研究提示：X射線照射引起的DNA斷裂性損傷是導(dǎo)致子宮內(nèi)膜胚胎接受性降低的重要原因之一。本研究為探討X射線照射與不孕癥的相關(guān)性提供實(shí)驗(yàn)依據(jù)。
[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.
【學(xué)位授予單位】：大連醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R818

【共引文獻(xiàn)】

相關(guān)期刊論文 前10條

1 梅梅;易杰;張疆_";;黏著斑激酶在整合素信號轉(zhuǎn)導(dǎo)中的研究進(jìn)展[J];重慶醫(yī)學(xué);2014年10期

2 劉建明;宋旭東;丁祺;;Skp2-siRNA干預(yù)對Eca-109食管癌細(xì)胞系p21和p27蛋白及細(xì)胞周期的影響[J];河北醫(yī)藥;2009年20期

3 劉波;陳宇鋒;鄭惠東;許貽斌;鄭盛華;鐘碩良;;醚菊酯對日本囊對蝦肝胰腺細(xì)胞DNA損傷效應(yīng)的研究[J];海洋科學(xué);2014年11期

4 劉波;許貽斌;鄭惠東;陳宇鋒;鄭盛華;鐘碩良;;氰戊菊酯對菲律賓蛤仔染毒后消化腺細(xì)胞的彗星實(shí)驗(yàn)分析[J];水產(chǎn)學(xué)報(bào);2014年05期

5 王繼慧;張小卿;馬月丹;張q

本文編號：1574350