内异症大鼠模型的建立及血管形成在其发病过程中的作用
发布时间：2018-08-28 07:14
【摘要】： 目的:子宫内膜异位症(endometriosis,EM)目前其发病机制不清,治疗效果不理想,复发率高达50%。在人类研究中由于有创性操作受到限制,因此需要动物模型作为研究的基础。在我国由于经济和社会的因素,大鼠常作为主要的动物模型。传统的实验方法是通过检查雌鼠阴道脱落细胞来选择有规律动情周期的大鼠,然后在其动情期进行实验,这一过程比较复杂,实验周期长。本文主要研究在不需确定大鼠动情周期的前提下,通过补充一定量的雌激素来支持异位灶的生长,建立大鼠自体移植模型,为EM的研究提供模型。 在EM病灶形成过程中,粘附-侵袭-血管形成是被多数学者认定的病理生理过程。异位灶的形成及维持需要血管建立来提供所需营养,在这一过程中有多种相关因子、酶的参与,有激素、免疫作用及局部微环境的影响。该实验通过检测大鼠在位及异位内膜中促血管形成因子的表达、腹腔微环境的改变,来研究大鼠模型的表现是否与人类相似,从而对大鼠模型做出正确的评价。 方法:将性成熟雌性SD大鼠的子宫角中段切下,使其内膜面朝向腹壁缝合到腹壁上。术后将大鼠随机分为两组,第8天开始给予不同剂量的雌激素,连续5天。术后4周再次剖腹,观察内膜生长情况;同时收集其腹腔冲洗液,用ELISA法检测肿瘤坏死因子(TNF-α)的表达情况;然后将成模与未成模的各分为两组,用免疫组化法检测大鼠在位及异位内膜血管内皮生长因子(VEGF)的表达。 结果:(1)给予高剂量雌激素组的成膜率为50%,低剂量雌激素组成模率为45%,差异没有统计学意义。(2)高剂量成模组腹腔冲洗液中TNF-α的含量为13.35±1.82pg/ml,未成模组的为8.80±2.02pg/ml;低剂量组中成模的为13.01±1.76pg/ml,未成模的为8.74±1.49pg/ml。高、低剂量组中成模都较未成模TNF-α的表达显著增高,差异有统计学意义;高剂量与低剂量各组之间相比没有显著性差异。(3)高剂量组中的成模组在位内膜VEGF的表达为1.22±0.59,异位内膜的为0.85±0.73,未成模组在位内膜VEGF的含量为0.59±0.40,其成模的在位和异位内膜都较未成模的在位内膜VEGF的表达增高,但差异没有统计学意义;低剂量组中的成模组在位内膜VEGF的表达为1.10±0.92,异位内膜的是1.10±0.92,未成模的在位内膜是0.30±0.33,同样成模的在位和异位内膜都较未成模的在位内膜VEGF的表达增高,且差异有统计学意义;高剂量与低剂量各组之间VEGF的表达没有显著性差异。 结论:(1)在不确定大鼠动情周期的情况下,通过术后补充雌激素,模型也可成功建立;同时过高剂量的雌激素对成模率没有影响。(2)TNF-α在EM的形成过程中起到了一定的促进作用,这与人类EM患者表现相似。(3)在位内膜VEGF的表达异常在EM形成过程中起关键作用,这与人类“在位内膜决定论”相似。总的来说大鼠模型可以作为研究EM的较好模型。
[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 伪 (TNF- 伪) 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- 伪 in peritoneal lavage fluid was 13.35 卤1.82 PG / ml in high dose model group, 8.80 卤2.02pg / ml in non-model group, and 8.80 卤2.02pg / ml in low dose group. The model-forming rate was 13.01 卤1.76 PG / ml, while that of the non-modular group was 8.74 卤1.49 pg / ml. The expression of TNF- 伪 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 卤0.59 in high dose group, 0.85 卤0.73 in ectopic endometrium, and 0.59 卤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 卤0.92 in low dose group, 1.10 卤0.92 in ectopic endometrium and 0.30 卤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- 伪 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.
【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R-332;R711.71
本文编号：2208675
[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 伪 (TNF- 伪) 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- 伪 in peritoneal lavage fluid was 13.35 卤1.82 PG / ml in high dose model group, 8.80 卤2.02pg / ml in non-model group, and 8.80 卤2.02pg / ml in low dose group. The model-forming rate was 13.01 卤1.76 PG / ml, while that of the non-modular group was 8.74 卤1.49 pg / ml. The expression of TNF- 伪 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 卤0.59 in high dose group, 0.85 卤0.73 in ectopic endometrium, and 0.59 卤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 卤0.92 in low dose group, 1.10 卤0.92 in ectopic endometrium and 0.30 卤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- 伪 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.
【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R-332;R711.71
【引证文献】
相关硕士学位论文 前1条
1 张颖;姜黄素抑制子宫内膜异位症模型大鼠血管形成的实验研究[D];湖北中医学院;2008年
，本文编号：2208675
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