【摘要】： 目的:子宮內(nèi)膜異位癥(endometriosis,EM)目前其發(fā)病機(jī)制不清,治療效果不理想,復(fù)發(fā)率高達(dá)50%。在人類(lèi)研究中由于有創(chuàng)性操作受到限制,因此需要?jiǎng)游锬Ｐ妥鳛檠芯康幕A(chǔ)。在我國(guó)由于經(jīng)濟(jì)和社會(huì)的因素,大鼠常作為主要的動(dòng)物模型。傳統(tǒng)的實(shí)驗(yàn)方法是通過(guò)檢查雌鼠陰道脫落細(xì)胞來(lái)選擇有規(guī)律動(dòng)情周期的大鼠,然后在其動(dòng)情期進(jìn)行實(shí)驗(yàn),這一過(guò)程比較復(fù)雜,實(shí)驗(yàn)周期長(zhǎng)。本文主要研究在不需確定大鼠動(dòng)情周期的前提下,通過(guò)補(bǔ)充一定量的雌激素來(lái)支持異位灶的生長(zhǎng),建立大鼠自體移植模型,為EM的研究提供模型。 在EM病灶形成過(guò)程中,粘附-侵襲-血管形成是被多數(shù)學(xué)者認(rèn)定的病理生理過(guò)程。異位灶的形成及維持需要血管建立來(lái)提供所需營(yíng)養(yǎng),在這一過(guò)程中有多種相關(guān)因子、酶的參與,有激素、免疫作用及局部微環(huán)境的影響。該實(shí)驗(yàn)通過(guò)檢測(cè)大鼠在位及異位內(nèi)膜中促血管形成因子的表達(dá)、腹腔微環(huán)境的改變,來(lái)研究大鼠模型的表現(xiàn)是否與人類(lèi)相似,從而對(duì)大鼠模型做出正確的評(píng)價(jià)。 方法:將性成熟雌性SD大鼠的子宮角中段切下,使其內(nèi)膜面朝向腹壁縫合到腹壁上。術(shù)后將大鼠隨機(jī)分為兩組,第8天開(kāi)始給予不同劑量的雌激素,連續(xù)5天。術(shù)后4周再次剖腹,觀察內(nèi)膜生長(zhǎng)情況;同時(shí)收集其腹腔沖洗液,用ELISA法檢測(cè)腫瘤壞死因子(TNF-α)的表達(dá)情況;然后將成模與未成模的各分為兩組,用免疫組化法檢測(cè)大鼠在位及異位內(nèi)膜血管內(nèi)皮生長(zhǎng)因子(VEGF)的表達(dá)。 結(jié)果:(1)給予高劑量雌激素組的成膜率為50%,低劑量雌激素組成模率為45%,差異沒(méi)有統(tǒng)計(jì)學(xué)意義。(2)高劑量成模組腹腔沖洗液中TNF-α的含量為13.35±1.82pg/ml,未成模組的為8.80±2.02pg/ml;低劑量組中成模的為13.01±1.76pg/ml,未成模的為8.74±1.49pg/ml。高、低劑量組中成模都較未成模TNF-α的表達(dá)顯著增高,差異有統(tǒng)計(jì)學(xué)意義;高劑量與低劑量各組之間相比沒(méi)有顯著性差異。(3)高劑量組中的成模組在位內(nèi)膜VEGF的表達(dá)為1.22±0.59,異位內(nèi)膜的為0.85±0.73,未成模組在位內(nèi)膜VEGF的含量為0.59±0.40,其成模的在位和異位內(nèi)膜都較未成模的在位內(nèi)膜VEGF的表達(dá)增高,但差異沒(méi)有統(tǒng)計(jì)學(xué)意義;低劑量組中的成模組在位內(nèi)膜VEGF的表達(dá)為1.10±0.92,異位內(nèi)膜的是1.10±0.92,未成模的在位內(nèi)膜是0.30±0.33,同樣成模的在位和異位內(nèi)膜都較未成模的在位內(nèi)膜VEGF的表達(dá)增高,且差異有統(tǒng)計(jì)學(xué)意義;高劑量與低劑量各組之間VEGF的表達(dá)沒(méi)有顯著性差異。 結(jié)論:(1)在不確定大鼠動(dòng)情周期的情況下,通過(guò)術(shù)后補(bǔ)充雌激素,模型也可成功建立;同時(shí)過(guò)高劑量的雌激素對(duì)成模率沒(méi)有影響。(2)TNF-α在EM的形成過(guò)程中起到了一定的促進(jìn)作用,這與人類(lèi)EM患者表現(xiàn)相似。(3)在位內(nèi)膜VEGF的表達(dá)異常在EM形成過(guò)程中起關(guān)鍵作用,這與人類(lèi)“在位內(nèi)膜決定論”相似。總的來(lái)說(shuō)大鼠模型可以作為研究EM的較好模型。
[Abstract]:Objective: at present, the pathogenesis of endometriosis (endometriosis,EM) is unclear, the therapeutic effect is not ideal, and the recurrence rate is as high as 50%. Because of the limitation of invasive operation in human research, animal models are needed as the basis of the research. In China, due to economic and social factors, rats are often used as the main animal model. The traditional experimental method is to select the rats with regular estrous cycle by examining the vaginal exfoliated cells of female rats, and then to conduct experiments in the estrus period. This process is complicated and the experimental period is long. This paper mainly studies the establishment of autotransplantation model of EM by adding a certain amount of estrogen to support the growth of ectopic foci without determining the estrous cycle of rats. Adhesion-invasion-angiogenesis is a pathophysiological process in the process of EM lesion formation. The formation and maintenance of ectopic foci require the establishment of blood vessels to provide the required nutrition. In this process, there are many related factors, the involvement of enzymes, hormones, immune function and local microenvironment. In this study, the expression of angiogenic factors and the changes of peritoneal microenvironment in eutopic and ectopic endometrium of rats were detected to study whether the performance of the rat model was similar to that of human, so as to make a correct evaluation of the rat model. Methods: the middle segment of the uterus horn of female SD rats was cut off and the endometrium was sutured to the abdominal wall towards the abdominal wall. Rats were randomly divided into two groups after operation. The rats were given different doses of estrogen on the 8th day for 5 days. Four weeks after operation, the growth of the intima was observed, the peritoneal lavage fluid was collected and the expression of tumor necrosis factor 偽 (TNF- 偽) was detected by ELISA method. The expression of vascular endothelial growth factor (VEGF) in eutopic and ectopic endometrium of rats was detected by immunohistochemistry. Results: (1) the rate of membrane formation was 50 in high dose estrogen group, and 45 in low dose estrogen group. (2) the content of TNF- 偽 in peritoneal lavage fluid was 13.35 鹵1.82 PG / ml in high dose model group, 8.80 鹵2.02pg / ml in non-model group, and 8.80 鹵2.02pg / ml in low dose group. The model-forming rate was 13.01 鹵1.76 PG / ml, while that of the non-modular group was 8.74 鹵1.49 pg / ml. The expression of TNF- 偽 in the high and low dose groups was significantly higher than that in the non-model group, and the difference was statistically significant. (3) the expression of VEGF in eutopic endometrium was 1.22 鹵0.59 in high dose group, 0.85 鹵0.73 in ectopic endometrium, and 0.59 鹵0.40 in non-model group. The expression of VEGF in eutopic endometrium was significantly higher than that in eutopic endometrium. The expression of VEGF in eutopic endometrium was 1.10 鹵0.92 in low dose group, 1.10 鹵0.92 in ectopic endometrium and 0.30 鹵0.33 in non-model eutopic endometrium. The expression of VEGF in eutopic and ectopic endometrium was higher than that in non-model eutopic endometrium. There was no significant difference in the expression of VEGF between high and low dose groups. Conclusion: (1) under the condition of uncertain estrous cycle, the model can be successfully established by supplement of estrogen after operation, and the rate of model formation is not affected by the high dose of estrogen. (2) TNF- 偽 plays a certain role in promoting the formation of EM. (3) abnormal expression of VEGF in eutopic endometrium plays a key role in the formation of EM, which is similar to human eutopic endometrium determinism. In general, the rat model can be used as a better model for studying EM.
【學(xué)位授予單位】：大連醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類(lèi)號(hào)】：R-332;R711.71

【引證文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 張穎;姜黃素抑制子宮內(nèi)膜異位癥模型大鼠血管形成的實(shí)驗(yàn)研究[D];湖北中醫(yī)學(xué)院;2008年

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本文編號(hào)：2208675