孕酮水平对小鼠子宫内膜蜕膜化的影响及其形态计算机三维重组

发布时间：2018-08-17 17:23

【摘要】： 子宫是亲代与后代个体之间生命延续的器官,作为一个生命孕育和最初生长的器官有着它一定的特性,例如月经的发生、与受精卵生长相适应的一系列变化等。子宫内膜的周期性变化主要由于垂体-卵巢轴的激素变化产生。随着卵泡的成熟、排卵,体内的雌、孕激素也发生着周期性的波动,雌、孕激素周期性的波动是月经现象产生的直接原因。由此可以看出,子宫是激素敏感器官,因此激素的不稳定就会使子宫产生异常的变化。在临床上,子宫内膜的异常出血是困扰妇产科临床和计划生育实践的重要临床现象。子宫内膜的异常出血是相对于正常的周期性出血而言的,因此,阐明一般周期性出血的成因对研究子宫内膜异常出血也具有一定意义。孕酮是雌性动物体内非常重要的性腺类固醇激素。既往的研究指出,孕酮是导致子宫周期性出血的重要因素。在整个月经周期中,多种激素发生波动,但孕酮的下降是导致子宫内膜蜕膜组织坏死崩解并出血的关键因素。既往实验实验主要是在模拟月经周期的激素水平波动,在实验的最终都会发生孕酮的撤退。但根据临床报告,大多数不规律出血的患者其体内孕酮水平并没有降低到月经周期分泌晚期的撤退水平,因此我们尝试在孕酮维持的状态下观察小鼠子宫内膜蜕膜的变化,这对于进一步阐明孕酮对于出血的影响有重要的意义。本实验用孕酮皮埋管维持小鼠体内的孕酮水平。首先确定皮埋管长度与所维持小鼠体内孕酮水平之间的关系。经测定,1cm长孕酮皮埋管所维持激素水平与小鼠妊娠第5天体内激素水平相当。所维持孕酮水平随皮埋管有效长度增长而升高,且存在较好的线性关系。这为我们后期就孕酮维持小鼠模型的建立做好准备。在小鼠月经模型的基础上,建立小鼠人工诱导蜕膜化孕酮水平维持模型。此模型与文献报告的三个模型的区别在于在小鼠蜕膜化模型的基础上维持孕酮水平,继而观察蜕膜的变化。我们根据文献所报告的方法,通过调节皮埋管长度使小鼠体内孕酮水平在实验观察阶段恒定于某一水平并连续监测阴道出血,观察孕酮水平与出血、出血时间的关系以及崩解后小鼠子宫内膜蜕膜组织的变化。结果提示,连续观察的120-248小时过程中,小鼠子宫内膜在维持孕酮的条件下仍然陆续发生了出血,组织切片显示各组普遍出现了蜕膜坏死和崩解,出血时间随孕酮水平的升高而延迟。随后我们观察了当孕酮水平维持在小鼠妊娠第5天水平下诱导蜕膜化后,不同时间的变化。我们分别观察了诱导蜕膜化后96、120、144小时的变化。随着观察时间的延长,组织坏死程度逐渐加深。在诱导后96小时,大体观察组织可看到蜕膜化区域呈结节样。切片观察可发现在宫腔中央有一空泡,蜕膜化组织中充满大量扩张血管。在诱导后144小时,坏死组织滞留于宫腔中且宫腔上皮未发生修复。实验中没有胚胎,仅通过宫腔注油给予宫腔上皮着床刺激,所观察到的变化为单纯的母体的变化。这些变化与妊娠子宫部分结构和变化相似。提示在妊娠过程中的一些变化与胚胎无关。在孕酮水平维持的过程中我们发现如果孕酮水平维持且蜕膜组织没有崩解时,子宫内膜中就会出现一些特殊的组织形态。在宫腔的中间会出现一个闭合的腔、闭合空腔周围空腔,以及几乎充满整个子宫的血窦。我们想获得这三种结构的空间构象。通过对组织连续切片并对其染色照相获得了在每个断层中三种结构的形态,后通过Adobe Photoshop以及Mimics软件对切片图像进行处理并进行三维重组,最终获得了三种结构空间构型及其相互空间关系,这对于阐明血管及其他结构在月经及妊娠过程中的作用提供了有价值的信息,也对形态学研究提供了一种新手段。本实验从组织形态学观察到最后的方法学研究都有一定的创新性,并且得到较明确的结果。希望可以对子宫内膜相关的研究提供有价值的信息。
[Abstract]:The uterus is the organ of life continuation between parent and offspring. As an organ of life gestation and initial growth, it has certain characteristics, such as the occurrence of menstruation, a series of changes adapted to the growth of fertilized eggs and so on. Maturity, ovulation, the body's estrogen, progesterone also occur periodic fluctuations, estrogen, progesterone periodic fluctuations are the direct cause of menstrual phenomena.
It can be seen from this that the uterus is a hormone sensitive organ, so the instability of hormones will cause abnormal changes in the uterus. Clinically, abnormal bleeding of the endometrium is an important clinical phenomenon in obstetrics and Gynecology and family planning practice. The cause of general periodic bleeding is also significant for studying abnormal bleeding of endometrium.
Progesterone is a very important gonadal steroid hormone in female animals. Previous studies have shown that progesterone is an important factor in causing periodic uterine bleeding. During the whole menstrual cycle, a variety of hormones fluctuate, but the decrease of progesterone is a key factor in causing endometrial and decidual tissue necrosis, disintegration and bleeding. But according to clinical reports, progesterone levels in most patients with irregular bleeding did not decrease to the level of withdrawal at the end of the menstrual cycle, so we tried to observe the endometrium of mice with progesterone maintenance. The change of decidua is important for further clarifying the effect of progesterone on bleeding.
The progesterone level in mice was maintained by a progesterone tube. First, the relationship between the length of the tube and the progesterone level in mice was determined. And there is a good linear relationship. This will prepare us for the establishment of progesterone maintenance mouse model in the later stage.
Based on the menstrual model of mice, a model of progesterone maintenance induced by artificial decidualization was established. The difference between this model and the three models reported in the literature was that progesterone levels were maintained on the basis of the mouse model of decidualization, and then the changes of decidua were observed. The progesterone level in mice was constant at a certain level during the experimental observation period and the vaginal bleeding was continuously monitored to observe the relationship between progesterone level and bleeding, bleeding time and the changes of endometrial Decidua Tissue after disintegration. Hemorrhage occurred successively. Tissue sections showed that decidual necrosis and disintegration were common in all groups. The time of bleeding was delayed with the increase of progesterone level.
Then we observed the changes of decidualization at different time points when progesterone levels were maintained at the 5th day of gestation in mice. We observed the changes at 96,120,144 hours after induction of decidualization. There was a vacuole in the center of the uterine cavity and a large number of dilated vessels were found in the decidualized tissue. At 144 hours after induction, the necrotic tissue was retained in the uterine cavity and the uterine epithelium was not repaired. Maternal changes. These changes are similar to some of the structures and changes of the pregnant uterus, suggesting that some changes during pregnancy are not related to the embryo.
During the maintenance of progesterone levels, we found that if progesterone levels were maintained and decidual tissue did not disintegrate, there would be some special histomorphology in the endometrium. In the middle of the uterine cavity there would be a closed cavity, a closed cavity around the cavity, and almost full of the uterine sinus. We wanted to obtain these three structures. Spatial conformation. Three kinds of structure shapes in each section were obtained by serial sections of tissue and staining photography. Then the slice images were processed and reconstructed by Adobe Photoshop and Mimics software. Finally, three kinds of structure spatial configuration and their spatial relationship were obtained. This is helpful to clarify the blood vessels and other knots. The role of structure in menstruation and pregnancy provides valuable information as well as a new means of morphological study.
We have observed from histomorphology that the final methodological study is innovative and has obtained definite results. We hope it can provide valuable information for endometrial-related research.
【学位授予单位】：中国协和医科大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R339.2

【参考文献】