巴多昔芬对子宫内膜异位症模型大鼠子宫内膜组织中ERα、ERβ、AP-1、VEGF表达的影响

发布时间：2018-04-24 21:43

本文选题：巴多昔芬 + 子宫内膜异位症　；参考：《郑州大学》2017年硕士论文

【摘要】：背景和目的子宫内膜异位症(endometriosis,EMs)是临床上常见的一种危害女性健康的慢性疾病,组织病理学上呈良性改变,但有很强的种植、侵袭、转移能力。有研究提示子宫内膜异位症的生物学行为可能源于雌激素暴露,作用于雌激素受体(estrogen receptor,ER),启动对应的信号通路,多条信号通路相互作用的结果促进了逆流的子宫内膜能在宫腔以外生长。目前临床上腹腔镜手术作为治疗子宫内膜异位症的金标准已经取得了很好的效果,但有时仍需辅助药物治疗,而用于治疗子宫内膜异位症的药物多在于减少内源性雌激素的产生,长期用药易导致机体低雌激素而出现一系列的围绝经期症状,而限制了这些药物的临床应用。有研究证明巴多昔芬作为第三代选择性雌激素受体调节剂(SERMs)能对大鼠EMs模型起到治疗作用并使雌激素受体(ER)的表达明显减少,因其表现出对大鼠EMs的治疗作用,而逐渐进入人们的视野,其临床应用前景广阔。本课题通过建立大鼠EMs模型,免疫组化法检测对照组大鼠子宫内膜、大鼠EMs模型(模型组、巴多昔芬组)在位子宫内膜和异位病灶中雌激素受体亚型α(ERα)、雌激素受体亚型β(ERβ)、AP-1组成成分c-jun蛋白、血管内皮生长因子(VEGF)表达情况,探讨ERα、ERβ、AP-1、VEGF在大鼠EMs模型中的表达特点及巴多昔芬治疗大鼠子宫内膜异位症的可能的作用机制。材料与方法1.材料河南省动物实验中心购买的清洁健康雌性未孕的SD大鼠100只,鼠龄60d~90d,质量220~250g,进入研究的各组(对照组、模型组、巴多昔芬组)大鼠的鼠龄、质量之间均无明显差异。2.方法观察雌性大鼠的正常发情周期,选取连续观察2个正常发情周期的大鼠进行建模。建模前一天均肌注雌激素,使大鼠处于发情期,皮下移植法建立子宫内膜异位症的动物模型:将剥除的子宫内膜剪为5×5mm大小的片段;采用皮下种植法将子宫内膜片段植入受体大鼠的腹肌和皮下筋膜之间,术后常规应用硫酸庆大霉素0.1ml肌肉注射预防感染3天。术后常规喂养3周,选取建模成功的56只大鼠(4只为无效造模)随机分为巴多昔芬组(3 mg·kg-1·d-1巴多昔芬灌胃,n=28)和模型组(每日等体积的生理盐水灌胃,n=28)。另外随机选取同一批次10只正常雌性大鼠作为对照组(喂养条件与术后实验组相同),4周后,在消毒、无菌条件下取模型组及巴多昔芬组异位病灶组织、在位子宫内膜,对照组子宫内膜分别固定于4%多聚甲醛,石蜡包埋,将蜡块连续切片,部分切片常规HE染色,部分用免疫组化方法检测ERα,ERβ,AP-1(c-jun蛋白)、VEGF的表达情况。结果1.对照组与模型组比较:(1)大鼠EMs模型在位内膜及异位病灶腺体、间质血管明显增加。(2)在大鼠EMs模型的异位病灶与对照组子宫内膜间ERα阳性率未见统计学差异(P=0.107),而其在在位内膜中的表达高于对照组(P0.01)。ERβ、c-jun、VEGF在模型组在位内膜及异位病灶中阳性率均高于对照组(P0.01),但其在位内膜及异位病灶中的表达未见明显差异。2.巴多昔芬组与模型组比较:巴多昔芬治疗后,巴多昔芬组与模型组相比,ERα的表达未表现出明显差异,但ERβ、AP-1、VEGF表达较模型组明显下降(P0.01)。结论1.ERα、ERβ、AP-1、VEGF在大鼠EMs模型中的表达特点支持“在位内膜决定论”。2.大鼠EMs模型在位内膜中ERα和ERβ均高表达,异位病灶中则仅表现为ERβ高表达,同时上调c-jun,VEGF的表达,ERβ可能在EMs的发生发展中起到较ERα更为重要的作用。3.巴多昔芬表现出对ERβ较强的的拮抗作用,并下调c-jun,VEGF的表达,这种表达模式的改变可能是巴多昔芬治疗大鼠EMs的一个重要机制。
[Abstract]:Background and objective endometriosis (EMs) is a clinically common chronic disease which is harmful to women's health. There is a benign pathological change in histopathology, but it has strong planting, invasion and metastasis. Estrogen receptor, ER), starting the corresponding signaling pathway, the results of multiple signal pathways interact to promote the reverse flow of endometrium outside the intrauterine growth. Clinical laparoscopic surgery as the gold standard for the treatment of endometriosis has achieved good results, but it is sometimes necessary to assist in the treatment of drugs, and for treatment. The drug of endometriosis is mainly due to the reduction of endogenous estrogen, which is easy to lead to a series of perimenopausal symptoms of low estrogen in the body, which restricts the clinical application of these drugs. It has been proved that baropoxifene as the third generation of selective estrogen receptor modulator (SERMs) can play a role in the EMs model of rats. The effect of the treatment and the expression of estrogen receptor (ER) is obviously reduced, because it shows the therapeutic effect of the rat EMs, and gradually enters the people's field of vision, and its clinical application is promising. In this subject, the rat endometrium of the control group was detected by the establishment of the rat EMs model and the immunohistochemical method was used in the rat EMs model (model group, barooxifene group). Estrogen receptor subtype alpha (ER alpha), estrogen receptor subtype beta (ER beta), AP-1 component c-jun protein and vascular endothelial growth factor (VEGF) expression in endometrium and ectopic lesions, and explore the expression of ER alpha, ER beta, AP-1, VEGF in rat EMs model and the possible mechanism of barooxifen in the treatment of endometriosis in rats. Materials and methods 1. SD rats were purchased from the animal experiment center of Henan Province, 100 rats of clean and healthy female unpregnant, rat age 60d~90d, and quality 220~250g. The rats of all groups (control group, model group, barooxifen group) had no obvious difference between the quality of the rats and the normal estrous cycle of female rats was observed by.2. method, and 2 continuous observation was selected. Rats in the normal oestrus cycle were modeled. A day before modeling, the rats were injected with estrogen to make the rats in the oestrus period. The animal model of endometriosis was established by subcutaneous transplantation: the stripped endometrium was cut into 5 x 5mm size fragments; the intrauterine fragment was implanted into the abdominal and subcutaneous fascia of the rat by subcutaneous implantation. For 3 days after operation, routine feeding of gentamicin sulfate 0.1ml was used to prevent infection for 3 days. After 3 weeks of routine feeding, 56 rats with successful modeling were randomly divided into barmoxifen group (3 mg. Kg-1. D-1 basoloxifen gavage, n=28) and model group (n=28). 10 normal female rats were used as the control group (the same as the control group). After 4 weeks, the ectopic tissue of the model group and batotecoxifene group were taken under disinfection and aseptic conditions. The endometrium was found in the endometrium. The endometrium in the control group was fixed to 4% polyformaldehyde and the paraffin was embedded. The paraffin blocks were sliced continuously, and some sections were stained with HE staining. Partial immunohistochemistry was used to detect the expression of ER alpha, ER beta, AP-1 (c-jun protein) and VEGF. Results the 1. control groups were compared with the model group: (1) the eutopic and ectopic lesion glands of the rat EMs model and the interstitial blood vessels were significantly increased. (2) there was no statistical difference between the ectopic lesion of the rat EMs model and the endometrium ER alpha positive rate in the control group (P=0.107). The expression in the eutopic endometrium (P0.01) was higher than that of the control group (P0.01).ER beta, and the positive rate of c-jun and VEGF in the eutopic and ectopic focus of the model group was higher than that of the control group (P0.01), but there was no significant difference in the expression of the eutopic and ectopic focus in the eutopic and ectopic focus group from the.2. barbadoxifen group and the model group: baruoxiifene group and the model group after treatment with barpecoxifene. The expression of ER beta, AP-1, and VEGF decreased significantly compared with that of the model group (P0.01). Conclusion 1.ER alpha, ER beta, AP-1, VEGF in the rat EMs model support the "eutectist of eutopic endometrium" in the intima of.2. rat EMs model. At the same time, the expression of c-jun, VEGF, ER beta may play a more important role in the development of EMs than ER alpha,.3. barbadoxifene shows strong antagonism to ER beta, and reduces the expression of c-jun and VEGF. The change of this expression pattern may be an important mechanism for the treatment of EMs in rats.

【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R711.71;R-332

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