药理性孕酮撤退建立小鼠月经模型及其初步研究
发布时间：2018-09-01 14:12
【摘要】： 子宫是一个独特的生殖器官,在雌激素和孕激素的序贯性作用下,子宫内膜形态和功能经历着周期性的特征性改变,包括子宫内膜的增殖、分化、在未受孕情况下的崩解以及相伴的出血等现象,构成了完整的月经周期。子宫内膜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。由于子宫异常出血是相对于规律性周期性出血而言的,所以,阐明子宫内膜周期性出血机制,是理解异常出血和发现应对策略的关键所在。模式动物模型的缺少是对其研究的主要限制因素之一。因此,建立小鼠月经样的改变并揭示月经发生的分子机制具有十分重要的意义。 1、成功建立了小鼠药理性孕酮撤退的月经模型。此模型与文献报告的两个模型的区别是孕酮撤退的方法不同。既往报告的方法是中止孕酮的供应,模拟周期晚期黄体衰退导致血中孕酮水平的下降。本研究在维持血清孕酮浓度的条件下,用米非司酮在受体水平阻断孕酮的作用,使蜕膜化的小鼠子宫出现崩解坏死及伴有出血的变化,模拟出与前述报告相同的月经生理现象。 2、进一步对本模型做了分子和生化表型的鉴定。对白细胞浸润、细胞凋亡、血管内皮生长因子及其受体和5个金属基质蛋白酶成员的表达和定位作了全时程动态变化的研究。结果表明,上述公认的月经相关分子和生化表达在小鼠药理性孕酮撤退的模型中变化与既往人和灵长类的月经研究所揭示的特征相符合。证明本研究建立的小鼠药理性孕酮撤退的月经模型系统在生理和生化特征上具备了月经模型的基本要素。为详细解析月经的分子机理提供了有价值的平台。 3、小鼠月经模型有别于灵长类的月经一些特征。由于宫腔内注入花生油导致的蜕膜化比较广泛,撤退孕酮出现的坏死崩解现象也更为典型和明显。不同时相的变化表现出明显的区域区别,特别是一些关键生化因子的分布出现显著的带状分布特征,且与崩解发生的区域相契合。这些表型特点不仅暗示了其中的相关分子机理,也为进一步的探索提供了有利的条件。 4、探讨了此模型孕酮撤退后的早期信号通路的变化。使用SuperArray信号转导通路发现者芯片对米非司酮处理0 h、8 h和16 h的变化作了初步探索。涉及18种信号通路的84个基因的表达。并对其中一个通路,NFκB信号通路在细胞水平做了初步验证和探讨,表明,NFκB在小鼠药理性孕酮撤退模型中参与了MMP3和MMP9的调控。
[Abstract]:The uterus is a unique reproductive organ. Under the sequential action of estrogen and progesterone, endometrial morphology and function undergo periodic characteristic changes, including endometrial proliferation and differentiation. Disintegration and associated bleeding in the absence of pregnancy constitute a complete menstrual cycle. Abnormal bleeding of endometrium is an important clinical phenomenon in obstetrics and gynecology and family planning practice. As abnormal uterine bleeding is relative to regular periodic bleeding, it is the key to understand abnormal bleeding and to discover coping strategies to elucidate the mechanism of endometrial periodic bleeding. The lack of model animal models is one of the main limiting factors in their research. Therefore, it is of great significance to establish menses like changes in mice and to reveal the molecular mechanism of menstruation. 1. The menstrual model of withdrawal of progesterone was successfully established. The difference between this model and the two reported models is the different method of progesterone withdrawal. Previously reported methods were to discontinue the supply of progesterone, and the late luteal decline in the simulated cycle resulted in a decrease in blood progesterone levels. In this study, under the condition of maintaining serum progesterone concentration, mifepristone was used to block the effect of progesterone at the receptor level, resulting in disintegration, necrosis and bleeding in decidualized mouse uterus. The same menstrual physiological phenomena as reported above were simulated. 2 the molecular and biochemical phenotypes of this model were further identified. WBC infiltration, apoptosis, expression and localization of vascular endothelial growth factor (VEGF) and its receptor and five metalloproteinases (MMPs) members were studied. The results showed that the changes of the known menstrual related molecules and biochemical expressions in the model of withdrawal of progesterone were consistent with the characteristics revealed by the menstrual studies of previous humans and primates. It is proved that the menstrual model system of the withdrawal of progesterone in this study has the basic physiological and biochemical characteristics of the menstrual model. It provides a valuable platform for explaining the molecular mechanism of menstruation in detail. 3. The menstrual model of mice is different from that of primates in some characteristics of menstruation. Due to the wide range of decidualization induced by intrauterine injection of peanut oil, the necrotic disintegration of retreating progesterone is more typical and obvious. The variation of different phases shows obvious regional differences, especially the distribution of some key biochemical factors shows a significant zonal distribution characteristic, which coincides with the region of disintegration. These phenotypic features not only imply the related molecular mechanism, but also provide favorable conditions for further exploration. 4. The changes of the early signal pathway after the withdrawal of progesterone in this model are discussed. The changes of mifepristone treated with mifepristone for 8 h and 16 h were studied using SuperArray signal transduction pathway discoverer chip. Expression of 84 genes involved in 18 signaling pathways. One of the pathways, NF 魏 B signaling pathway, was preliminarily verified and explored at the cell level, indicating that NF 魏 B was involved in the regulation of MMP3 and MMP9 in the withdrawal model of progesterone.
【学位授予单位】：中国协和医科大学
【学位级别】：博士
【学位授予年份】：2008
【分类号】：R-332
本文编号：2217449
[Abstract]:The uterus is a unique reproductive organ. Under the sequential action of estrogen and progesterone, endometrial morphology and function undergo periodic characteristic changes, including endometrial proliferation and differentiation. Disintegration and associated bleeding in the absence of pregnancy constitute a complete menstrual cycle. Abnormal bleeding of endometrium is an important clinical phenomenon in obstetrics and gynecology and family planning practice. As abnormal uterine bleeding is relative to regular periodic bleeding, it is the key to understand abnormal bleeding and to discover coping strategies to elucidate the mechanism of endometrial periodic bleeding. The lack of model animal models is one of the main limiting factors in their research. Therefore, it is of great significance to establish menses like changes in mice and to reveal the molecular mechanism of menstruation. 1. The menstrual model of withdrawal of progesterone was successfully established. The difference between this model and the two reported models is the different method of progesterone withdrawal. Previously reported methods were to discontinue the supply of progesterone, and the late luteal decline in the simulated cycle resulted in a decrease in blood progesterone levels. In this study, under the condition of maintaining serum progesterone concentration, mifepristone was used to block the effect of progesterone at the receptor level, resulting in disintegration, necrosis and bleeding in decidualized mouse uterus. The same menstrual physiological phenomena as reported above were simulated. 2 the molecular and biochemical phenotypes of this model were further identified. WBC infiltration, apoptosis, expression and localization of vascular endothelial growth factor (VEGF) and its receptor and five metalloproteinases (MMPs) members were studied. The results showed that the changes of the known menstrual related molecules and biochemical expressions in the model of withdrawal of progesterone were consistent with the characteristics revealed by the menstrual studies of previous humans and primates. It is proved that the menstrual model system of the withdrawal of progesterone in this study has the basic physiological and biochemical characteristics of the menstrual model. It provides a valuable platform for explaining the molecular mechanism of menstruation in detail. 3. The menstrual model of mice is different from that of primates in some characteristics of menstruation. Due to the wide range of decidualization induced by intrauterine injection of peanut oil, the necrotic disintegration of retreating progesterone is more typical and obvious. The variation of different phases shows obvious regional differences, especially the distribution of some key biochemical factors shows a significant zonal distribution characteristic, which coincides with the region of disintegration. These phenotypic features not only imply the related molecular mechanism, but also provide favorable conditions for further exploration. 4. The changes of the early signal pathway after the withdrawal of progesterone in this model are discussed. The changes of mifepristone treated with mifepristone for 8 h and 16 h were studied using SuperArray signal transduction pathway discoverer chip. Expression of 84 genes involved in 18 signaling pathways. One of the pathways, NF 魏 B signaling pathway, was preliminarily verified and explored at the cell level, indicating that NF 魏 B was involved in the regulation of MMP3 and MMP9 in the withdrawal model of progesterone.
【学位授予单位】：中国协和医科大学
【学位级别】：博士
【学位授予年份】：2008
【分类号】：R-332
【引证文献】
相关硕士学位论文 前1条
1 孙琦;活血方与活血加味方对小鼠月经模型干预作用及与阿魏酸煎出量的相关性实验研究[D];河北医科大学;2012年
，本文编号：2217449
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