本文選題：子宮內(nèi)膜異位癥 + 缺氧��； 參考：《中國人民解放軍醫(yī)學(xué)院》2014年博士論文

【摘要】：背景子宮內(nèi)膜異位癥（endometriosis，EMs）是一種現(xiàn)代婦科病，在育齡期女性的發(fā)病率為6-10%，在不孕不育女性中可達(dá)30%。EMs患者中卵巢子宮內(nèi)膜樣癌和透明細(xì)胞癌發(fā)病風(fēng)險(xiǎn)明顯增高。目前，腹腔鏡手術(shù)與性激素的應(yīng)用已成為EMs主要治療方法，輔助生殖技術(shù)為患者改善生育狀況帶來希望，然而術(shù)后復(fù)發(fā)、粘連引起的盆腔疼痛及不孕，甚至遠(yuǎn)處轉(zhuǎn)移及癌變嚴(yán)重?fù)p害著女性身心健康。 1927年Sampson提出的經(jīng)血逆流種植學(xué)說至今仍被眾多實(shí)驗(yàn)證據(jù)所支持，但經(jīng)血逆流是婦女月經(jīng)期常見的現(xiàn)象，而EMs發(fā)生率與其比較相對較低。近年來EMs“在位內(nèi)膜決定論”的發(fā)病學(xué)說補(bǔ)充和發(fā)展了經(jīng)血逆流理論。但是在位子宮內(nèi)膜究竟通過什么途徑“決定”了EMs何時發(fā)生及如何發(fā)展，尚無相關(guān)報(bào)道。EMs直接與月經(jīng)相關(guān)，月經(jīng)期間子宮螺旋動脈收縮，子宮內(nèi)膜處于缺血、缺氧狀態(tài)，內(nèi)膜碎片隨經(jīng)血逆流至盆腔，在異位粘附、侵襲、血管形成之前均處于缺氧狀態(tài)。近年研究發(fā)現(xiàn)缺氧影響著腫瘤的發(fā)生、發(fā)展；而EMs可以浸潤、轉(zhuǎn)移及復(fù)發(fā)，有類似惡性腫瘤的特性和生物學(xué)行為。因此，探索缺氧在EMs發(fā)生發(fā)展中的可能作用機(jī)制，對于解釋EMs諸多生物學(xué)行為根源，尋找新的診斷、治療和預(yù)防措施具有一定的理論和實(shí)踐意義。 目的研究持續(xù)缺氧以及急性缺氧對EMs細(xì)胞及組織增生，凋亡，血管生成等影響；探討不同來源的子宮在位、異位內(nèi)膜組織，，以及在SCID小鼠模型的不同移植部位，EMs病灶形成與缺氧、血管生成和粘連等相關(guān)因子的關(guān)系。 方法原代培養(yǎng)人子宮內(nèi)膜細(xì)胞，比較缺氧或者常氧培養(yǎng)對于不同來源子宮內(nèi)膜細(xì)胞增殖凋亡的影響；通過SCID小鼠異種移植人子宮內(nèi)膜建立EMs在體模型，觀察經(jīng)缺氧或高氧預(yù)處理移植物后病灶的生長情況；在SCID小鼠EMs模型觀察EMs在位或異位內(nèi)膜移植，以及SCID小鼠腹腔內(nèi)或皮下不同部位EMs移植物的生長情況。應(yīng)用細(xì)胞熒光免疫方法、流式細(xì)胞儀分析，HE染色，免疫組化技術(shù)，Western blot和ELISA等方法檢測與增殖，凋亡，血管生成和缺氧等相關(guān)蛋白因子的表達(dá)。 結(jié)果1、子宮內(nèi)膜細(xì)胞在缺氧培養(yǎng)組比常氧培養(yǎng)組凋亡率上升；在缺氧環(huán)境下，EMs細(xì)胞偽足比對照組細(xì)胞形成增多，而凋亡率減少。缺氧培養(yǎng)使EMs細(xì)胞表達(dá)HIF-1α，VEGF，Ki67比常氧對照組增加。2、在SCID小鼠EMs模型中，經(jīng)缺氧預(yù)處理后的移植物與高氧及常氧預(yù)處理對照組比較，移植物重量增加；缺氧預(yù)處理組病灶中的MVD、HIF-1α、VEGF和Ki67等蛋白表達(dá)增加。3、不同來源的內(nèi)膜組織在SCID小鼠腹腔內(nèi)生長方式不同，EMs異位內(nèi)膜移植物的重量，組織中CD34、VEGF、HIF-1α和CA125表達(dá)以及SCID小鼠腹水中CA125含量比在位內(nèi)膜組以及對照組內(nèi)膜移植物明顯增加。4、SCID小鼠腹腔內(nèi)移植組織與網(wǎng)膜及腹膜形成粘連帶，粘連帶內(nèi)可見血管生成；而皮下移植的組織粘連帶形成相對較少。皮下移植病灶的體積增加明顯，以及MMP-2和TIMP-2，MVD，VEGF和HIF-1α表達(dá)均高于腹腔內(nèi)移植病灶。 結(jié)論1、缺氧培養(yǎng)促使EMs內(nèi)膜細(xì)胞凋亡的同時，也促使EMs內(nèi)膜細(xì)胞表達(dá)HIF-1α、血管生成和增生相關(guān)蛋白；EMs細(xì)胞抗凋亡以及增生能力要高于對照內(nèi)膜細(xì)胞。2、急性缺氧促進(jìn)EMs移植病灶表達(dá)HIF-1α、血管生成以及增生相關(guān)因子，影響病灶生長。3、EMs異位內(nèi)膜移植病灶生長能力要強(qiáng)于EMs在位內(nèi)膜以及非EMs對照組內(nèi)膜， EMs在位內(nèi)膜與對照內(nèi)膜也存在差別，異位內(nèi)膜細(xì)胞可能是EMs在位內(nèi)膜缺氧選擇的結(jié)果。4、SCID小鼠不同部位的EMs移植物生長以及粘連帶形成存在差異，粘連、血管生成以及HIF-1α等因子表達(dá)也不同，這說明局部缺氧微環(huán)境影響EMs病灶生物學(xué)行為。
[Abstract]:Background endometriosis (EMs) is a modern gynecologic disease. The incidence of women in the childbearing age is 6-10%. The risk of ovarian endometrioid and transparent cell carcinoma in the patients with infertile women is significantly higher than that of 30%.EMs patients. At present, the application of laparoscopic hand and sex hormone has become the main treatment of EMs. Assisted reproductive technology brings hope for the patients to improve their fertility. However, postoperative recurrence, pelvic pain and infertility caused by adhesion, even distant metastasis and canceration seriously damage the physical and mental health of women.
The theory of reflux planting for blood flow, proposed by Sampson in 1927, is still supported by numerous experimental evidence, but the reverse flow of blood is a common phenomenon in women's menstrual period, and the incidence of EMs is relatively low. In recent years, the theory of EMs "eutopic endometrial determinism" has been supplemented and developed by the theory of reverse flow of blood, but the eutopic endometrium is in fact By what means "determines" when and how EMs develops, there is no related reports that.EMs is directly related to menstruation, the uterine spiral artery contraction, the endometrium in the ischemic and anoxic state, the endometrium fragments with the menstrual flow to the pelvic cavity, in the anoxic state before ectopic adhesion, invasion and angiogenesis. Anoxia affects the occurrence and development of tumor, and EMs can infiltrate, metastases and relapse with the characteristics and biological behavior of malignant tumor. Therefore, it is possible to explore the possible mechanism of hypoxia in the development of EMs, and to explain the root of the biological behavior of EMs and to find new diagnosis, treatment and prevention measures. Practice meaning.
Objective to study the effects of continuous hypoxia and acute hypoxia on the proliferation, apoptosis and angiogenesis of EMs cells and tissues, and to explore the relationship between different sources of uterus, ectopic endometrium, different sites of transplantation of SCID mice, the formation of EMs focus and the related factors of hypoxia, angiogenesis and adhesion.
Methods the primary endometrium cells were cultured, and the effects of hypoxia or aerobia on the proliferation and apoptosis of endometrium cells in different sources were compared. EMs in the endometrium of the xenotransplantation of SCID mice was established to observe the growth of the lesion after the hypoxia or hyperoxia preconditioning of the graft, and the EMs in the EMs model of SCID mice was observed. The growth of EMs grafts in the abdominal or subcutaneous parts of SCID mice and the growth of transplanting or ectopic endometrium, and using cell fluorescence immunoassay, flow cytometry, HE staining, immunohistochemistry, Western blot and ELISA, were used to detect the expression of protein factors, such as proliferation, apoptosis, angiogenesis and hypoxia.
Results 1, the apoptosis rate of the endometrium cells in the hypoxia culture group was higher than that in the normal oxygen culture group. In the hypoxia environment, the cell formation of EMs cell hypoplastic cells increased and the apoptosis rate decreased. The hypoxia culture made EMs cells express HIF-1 alpha, VEGF, Ki67 more.2 than the normal oxygen control group. In the EMs model of SCID mice, the anoxic pretreated graft was found in the EMs model of SCID mice. Compared with hyperoxia and hyperoxia preconditioning group, the weight of graft increased, and the expression of MVD, HIF-1 a, VEGF and Ki67 increased.3 in the focus of the hypoxia preconditioning group. The growth patterns of endometrium in the abdominal cavity of SCID mice were different in different sources, the weight of EMs endometrium graft, CD34, VEGF, HIF-1 alpha and CA125 expression in the tissue and the small SCID. The content of CA125 in rat ascites was significantly increased by.4 than in the eutopic endometrium and the control group. The intraperitoneal transplantation tissue in SCID mice was visially connected with the omentum and peritoneum, and the vascular formation was visible in the adhesion zone; and the tissue adhesion zone formed by subcutaneous transplantation was relatively small. The volume of subcutaneous transplantation lesions increased significantly, and MMP-2 and TIMP-2, MV The expressions of D, VEGF and HIF-1 alpha were all higher than those in abdominal cavity.
Conclusion 1, hypoxia culture promotes the apoptosis of EMs intima cells, and also promotes the expression of HIF-1 alpha, angiogenesis and proliferation related proteins in EMs endometrium cells. The anti apoptosis and proliferation ability of EMs cells is higher than that of the control endometrial cells.2. Acute hypoxia promotes the expression of HIF-1 a in EMs transplant lesions, angiogenesis and proliferation related factors, which affect the growth of the lesion. The growth ability of.3, EMs ectopic endometrium transplantation focus is stronger than that of EMs eutopic endometrium and non EMs control group. EMs eutopic endometrium and control endometrium are also different. Ectopic endometrium cells may be the result of EMs eutopic endometrial anoxia selection,.4, EMs graft growth and adhesion formation in different parts of SCID mice, adhesions, and blood vessels The expression of HIF-1 and alpha is also different. This indicates that the local hypoxia microenvironment affects the biological behavior of EMs lesions.

【學(xué)位授予單位】：中國人民解放軍醫(yī)學(xué)院
【學(xué)位級別】：博士
【學(xué)位授予年份】：2014
【分類號】：R711.71

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