卵巢型子宫内膜异位症大鼠模型的建立与评价
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河北中医药大学,河北省中西医结合肝肾病证研究重点实验室, 石家庄 050091

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Establishment and evaluation of a rat model of ovarian endometriosis
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Hebei Integrated Traditional Chinese and Western Medicine Key Laboratory of Liver and Kidney Diseases, Hebei University of Traditional Chinese Medicine, Shijiazhuang 050091, China

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    摘要:

    目的 采用子宫角返折法建立卵巢型子宫内膜异位症(endometriosis,EMS)大鼠模型,评估子宫角返折法建立卵巢型EMS大鼠模型的可行性,为探究EMS发病机制和治疗方法等提供理想的动物模型。 方法 将50只SPF级雌性SD大鼠随机分为5组,分别为假手术组、术后1周组、术后2周组、术后3周组、术后4周组,每组10只,除假手术组外采用子宫角返折法建立卵巢型EMS模型,分别在术后1、2、3、4周观察造模成功率、异位灶体积及质量,初步评价模型。采用苏木素-伊红(HE)染色观察异位灶形态,免疫组化法检测子宫及异位灶组织中增殖细胞核抗原(PCNA)、细胞增殖核抗原(Ki67)、上皮细胞钙黏蛋白(E-cadherin)、神经钙黏蛋白(N-cadherin)的表达,进一步评价模型并分析术后不同时间点异位灶细胞增殖及上皮间质转化程度。 结果 术后1、2、3、4周组造模成功率分别为80%、90%、100%、100%(P>0.05)。与术后1、2周组相比,术后3、4周组异位灶体积明显增大,质量明显增加(P<0.01)。HE染色可见异位灶组织中有子宫内膜上皮细胞、间质细胞和少量腺体。免疫组织化学染色结果显示,与假手术组相比,术后1、2、3、4周组大鼠子宫组织中Ki67、PCNA、N-cadherin表达水平显著升高(P<0.05,P<0.01);E-cadherin表达水平显著降低(P<0.05,P<0.01)。与术后1周组相比,术后2、3、4周组大鼠子宫及异位灶组织中Ki67、PCNA、N-cadherin表达水平显著升高(P<0.05,P<0.01);E-cadherin表达水平显著降低(P<0.05,P<0.01)。 结论 通过子宫角返折法可以建立大鼠卵巢型EMS的模型,术后1周可以观察到异位灶形成,造模成功率随造模时间延长而提高,于术后3周趋于稳定。Ki67、PCNA、N-cadherin在子宫及异位灶组织中均有显著性表达,表达强度随造模时间延长而递增。E-cadherin在子宫及异位灶组织中的表达强度随造模时间延长而降低。结果表明,子宫角返折法构建EMS大鼠模型成模率高,可作为一种稳定的EMS建模方法。

    Abstract:

    Objective To establish a rat model of ovarian endometriosis (EMS) using the horn reversion method, to provide an ideal animal model for exploring the pathogenesis and treatment of EMS. Methods Fifty SPFgrade female SD rats were divided randomly into five groups: a sham group, and 1, 2, 3, and 4 weeks after surgery groups, respectively (n=10 rats per group). Apart from the sham group, an ovarian-type EMS model was established in the other groups by the uterine horn refracture method, and the modeling success rate, and ectopic foci volume and mass were observed in each group. The morphology of ectopic foci was observed by hematoxylin-eosin (HE), and expression of proliferating cell nuclear antigen (PCNA), Ki67, epithelial cadherin (E-cadherin), and neural cadherin (N-cadherin) in the uterus and ectopic foci tissues were detected by immunohistochemistry. The model was further evaluated and the degree of cell proliferation and epithelial mesenchymal transformation at different times after modeling were analyzed. Results The modeling success rates in the 1, 2, 3, and 4 weeks after surgery groups were 80%, 90%, 100%, and 100%, respectively (P>0.05). The volume and mass of the ectopic foci were significantly greater in the 3 and 4 weeks after surgery groups compared with the 1 and 2 weeks after surgery groups (P<0.01). HE staining showed endometrial epithelial cells, mesenchymal cells and a few glands in the ectopic foci tissues. Immunohistochemical staining showed that expression levels of Ki67, PCNA, and N-cadherin in uterus tissues were significantly higher (P<0.05, P<0.01) in all the model groups compared with the sham group, while expression levels of E-cadherin were significantly lower (P<0.05, P<0.01). Expression levels of Ki67, PCNA, and N-cadherin in the uterus and ectopic foci tissues were significantly higher (P<0.05, P<0.01) in the 2, 3, and 4 weeks after surgery groups compared with the 1 week after surgery group, while expression levels of E-cadherin were significantly lower (P<0.05, P<0.01). Conclusions The uterine horn reversion method can be used to establish an ovarian EMS model in rats. Ectopic lesions can be observed 1 week after surgery. The success rate of modeling increases with modeling time, stabilizing at 3 weeks postoperatively. Ki67, PCNA, and N-cadherin were significantly expressed in the uterus and ectopic foci tissues, and their expression levels increased with modeling time, while E-cadherin expression in the uterus and ectopic foci tissues decreased with modeling time. These Results showed good modeling success of the EMS rat model, suggesting that it could be used as a stable EMS modeling method.

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马艺鸣,刘慧敏,孟鑫,齐嘉泽,安铭立,付欣平,陈景伟.卵巢型子宫内膜异位症大鼠模型的建立与评价[J].中国实验动物学报,2025,33(7):947~957.

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  • 收稿日期:2024-12-11
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