大鼠薄型子宫内膜模型的建立和鉴定

发布时间：2018-07-03 17:55

本文选题：薄型子宫内膜 + 无水乙醇　；参考：《中南大学》2011年硕士论文

【摘要】：目的：探索宫腔注射无水乙醇建立大鼠薄型子宫内膜实验模型的方法,并鉴定模型是否成立。方法：取性成熟未交配雌性具有动情周期的Sprague-Dawley(SD)大鼠25只,随机分为两组：对照组5只(10个子宫),实验组20只(40个子宫)。对照组自宫角处注入生理盐水,贮留5分钟,实验组自宫角处注入95%无水乙醇,分别贮留5分钟、10分钟。根据液体贮留时间不同,实验组随机分为5分钟组、10分钟组,各10只大鼠(20个子宫)。经过两个动情周期后,取处于动情期大鼠的子宫,测量子宫内膜厚度：若实验组子宫内膜厚度与对照组相比无变化或者子宫内膜层完全坏死,则造模失败。若子宫内膜变薄,我们将内膜变薄的子宫归为造模组,对其HE染色后子宫内膜组织形态学进行进一步观察,并采用免疫组织化学法方法检测角蛋白、波形蛋白、VEGF、ERα的表达,分析子宫内膜形态学特点、子宫内膜上皮细胞和间质中细胞再生长情况以及VEGF、ERα表达情况是否符合薄型内膜的特点,从而鉴定模型是否成功建立。结果： 1.宫腔注射95%无水乙醇经过两个动情周期后,5分钟组中,1只大鼠死亡,4个子宫的内膜厚度恢复至正常,14个子宫的内膜变薄；10分钟组中,2只大鼠死亡,8只大鼠子宫内膜层坏死或者子宫全层坏死。 2.与对照组相比：造模组子宫壁全层明显变薄,子宫内膜层(764.1±100.2μmvs 234.6±48.1μm,p0.01)较肌层更明显,且未见明显波浪形；腺体稀疏,部分腺腔膨大成囊状,腺上皮及腔上皮细胞呈低柱、立方甚至扁平状；血管减少,部分管腔膨大。10分钟组子宫内膜层大部分为无细胞结构区域,仅存少数细胞结构轮廓；浅肌层甚至子宫全层坏死,肌层血管扩张,内可见血栓形成。 3.造模组单位内膜面积中角蛋白面积小于对照组(11.46±2.40%vs 17.82±2.28%,p0.05),差异有统计学意义；造模组单位间质面积中波形蛋白面积小于对照组(8.30±2.84%vs 13.62±3.18%,p0.05),差异有统计学意义；造模组子宫内膜上皮细胞和间质细胞中VEGF平均光密度低于对照组(0.033±0.0053 vs 0.054±0.0084,p0.05),差异有统计学意义。造模组子宫内膜上皮细胞中ERa组织学积分稍低于对照组(3.12±0.07 vs 3.20±0.12,p0.05),差异无统计学意义。结论：初步证实95%无水乙醇0.5ml宫腔贮留5分钟可以成功形成大鼠薄型子宫内膜,造模成功率为70%。
[Abstract]:Aim: to explore the method of establishing rat model of thin endometrium by intrauterine injection of anhydrous alcohol and to identify whether the model was established. Methods: Twenty-five Sprague-Dawley (SD) rats with oestrous cycle were randomly divided into two groups: control group (n = 5) (n = 10) and experimental group (n = 20) (n = 40). The control group was injected with normal saline from the angle of uterus for 5 minutes, and the experimental group was injected with 95% anhydrous alcohol from the corner of the uterus for 5 minutes and 10 minutes respectively. According to the time of liquid storage, the experimental group was randomly divided into 5 minute group and 10 minutes group, each 10 rats (20 uteri). After two estrus cycles, the endometrium thickness of rats in estrus period was measured. If the endometrial thickness of the experimental group was not changed compared with the control group or the endometrium was completely necrotic, the model failed. If the endometrium became thinner, we classified the thinned endometrium as a model group, observed the morphology of endometrium after HE staining, and detected the expression of keratin, vimentin and VEGFER 伪 by immunohistochemical method. The morphological characteristics of endometrium, the growth of endometrial epithelial cells and stromal cells, and the expression of VEGF ER 伪 were analyzed. Results: 1. After two estrous cycles, one rat died, four endometrium thickness returned to normal, and 14 endometrium became thinner in uterine cavity injection of 95% anhydrous alcohol after two estrous cycles. In the 10-minute group, 2 rats died and 8 rats died of endometrial necrosis or full uterine necrosis. 2. Compared with the control group, the endometrium layer (764.1 卤100.2 渭 mvs 234.6 卤48.1 渭 m P0.01) was significantly thinner than that of the control group (764.1 卤100.2 渭 mvs 234.6 卤48.1 渭 m P0.01), and there was no obvious wavy shape in the endometrial layer, and the glands were sparse, some of the glandular lumens were cystic, the glandular epithelium and the lumen epithelial cells were low columnar. Cuboidal or even flattened; reduced blood vessels. The endometrial layer of partial lumen bulging for 10 minutes was mostly cell-free, with only a few cell contours; superficial muscle layer and even the whole uterine layer were necrotic, myometrium vasodilated. Intrinsic thrombus formation. 3. The median area of keratin per unit intima in the model group was smaller than that in the control group (11.46 卤2.40%vs 17.82 卤2.28), and the area of vimentin in the interstitial area of the model group was lower than that in the control group (8.30 卤2.84%vs 13.62 卤3.18p0.05). The average optical density in endometrial epithelial cells and stromal cells in model group was significantly lower than that in control group (0.033 卤0.0053 vs 0.054 卤0.0084 p0.05). The ERa histological score in endometrial epithelial cells in the model group was slightly lower than that in the control group (3.12 卤0.07 vs 3.20 卤0.12p0.05). Conclusion: 95% anhydrous ethanol 0.5ml uterine cavity storage for 5 minutes can successfully form rat thin endometrium, and the success rate of modeling is 70.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R-332

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