藥理性孕酮撤退建立小鼠月經(jīng)模型及其初步研究
發(fā)布時(shí)間：2018-09-01 14:12
【摘要】： 子宮是一個(gè)獨(dú)特的生殖器官,在雌激素和孕激素的序貫性作用下,子宮內(nèi)膜形態(tài)和功能經(jīng)歷著周期性的特征性改變,包括子宮內(nèi)膜的增殖、分化、在未受孕情況下的崩解以及相伴的出血等現(xiàn)象,構(gòu)成了完整的月經(jīng)周期。子宮內(nèi)膜異常出血是困擾婦產(chǎn)科臨床和計(jì)劃生育實(shí)踐的重要臨床現(xiàn)象。由于子宮異常出血是相對(duì)于規(guī)律性周期性出血而言的,所以,闡明子宮內(nèi)膜周期性出血機(jī)制,是理解異常出血和發(fā)現(xiàn)應(yīng)對(duì)策略的關(guān)鍵所在。模式動(dòng)物模型的缺少是對(duì)其研究的主要限制因素之一。因此,建立小鼠月經(jīng)樣的改變并揭示月經(jīng)發(fā)生的分子機(jī)制具有十分重要的意義。 1、成功建立了小鼠藥理性孕酮撤退的月經(jīng)模型。此模型與文獻(xiàn)報(bào)告的兩個(gè)模型的區(qū)別是孕酮撤退的方法不同。既往報(bào)告的方法是中止孕酮的供應(yīng),模擬周期晚期黃體衰退導(dǎo)致血中孕酮水平的下降。本研究在維持血清孕酮濃度的條件下,用米非司酮在受體水平阻斷孕酮的作用,使蛻膜化的小鼠子宮出現(xiàn)崩解壞死及伴有出血的變化,模擬出與前述報(bào)告相同的月經(jīng)生理現(xiàn)象。 2、進(jìn)一步對(duì)本模型做了分子和生化表型的鑒定。對(duì)白細(xì)胞浸潤、細(xì)胞凋亡、血管內(nèi)皮生長因子及其受體和5個(gè)金屬基質(zhì)蛋白酶成員的表達(dá)和定位作了全時(shí)程動(dòng)態(tài)變化的研究。結(jié)果表明,上述公認(rèn)的月經(jīng)相關(guān)分子和生化表達(dá)在小鼠藥理性孕酮撤退的模型中變化與既往人和靈長類的月經(jīng)研究所揭示的特征相符合。證明本研究建立的小鼠藥理性孕酮撤退的月經(jīng)模型系統(tǒng)在生理和生化特征上具備了月經(jīng)模型的基本要素。為詳細(xì)解析月經(jīng)的分子機(jī)理提供了有價(jià)值的平臺(tái)。 3、小鼠月經(jīng)模型有別于靈長類的月經(jīng)一些特征。由于宮腔內(nèi)注入花生油導(dǎo)致的蛻膜化比較廣泛,撤退孕酮出現(xiàn)的壞死崩解現(xiàn)象也更為典型和明顯。不同時(shí)相的變化表現(xiàn)出明顯的區(qū)域區(qū)別,特別是一些關(guān)鍵生化因子的分布出現(xiàn)顯著的帶狀分布特征,且與崩解發(fā)生的區(qū)域相契合。這些表型特點(diǎn)不僅暗示了其中的相關(guān)分子機(jī)理,也為進(jìn)一步的探索提供了有利的條件。 4、探討了此模型孕酮撤退后的早期信號(hào)通路的變化。使用SuperArray信號(hào)轉(zhuǎn)導(dǎo)通路發(fā)現(xiàn)者芯片對(duì)米非司酮處理0 h、8 h和16 h的變化作了初步探索。涉及18種信號(hào)通路的84個(gè)基因的表達(dá)。并對(duì)其中一個(gè)通路,NFκB信號(hào)通路在細(xì)胞水平做了初步驗(yàn)證和探討,表明,NFκB在小鼠藥理性孕酮撤退模型中參與了MMP3和MMP9的調(diào)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.
【學(xué)位授予單位】：中國協(xié)和醫(yī)科大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2008
【分類號(hào)】：R-332
本文編號(hào)：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.
【學(xué)位授予單位】：中國協(xié)和醫(yī)科大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2008
【分類號(hào)】：R-332
【引證文獻(xiàn)】
相關(guān)碩士學(xué)位論文 前1條
1 孫琦;活血方與活血加味方對(duì)小鼠月經(jīng)模型干預(yù)作用及與阿魏酸煎出量的相關(guān)性實(shí)驗(yàn)研究[D];河北醫(yī)科大學(xué);2012年
，本文編號(hào)：2217449
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