子宫内膜异位症纤维化小鼠模型构建方法及其应用
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1.上海中医药大学,上海 201203; 2. 海军军医大学附属长海医院,上海 200433

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Establishment and application of a mouse model of endometriosis with fibrosis
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1.Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China. 2. Changhai Hospital, Naval Military Medical University, Shanghai 200433

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    摘要:

    目的 探索一种高效的子宫内膜异位症纤维化小鼠模型建立方法及其评价方法。 方法 改良的腹腔注射法构建 BALB/ c 小鼠子宫内膜异位症纤维化模型;收集小鼠腹腔灌洗液,ELISA 试剂盒检测灌洗液 TGF-β1 水平;对小鼠腹腔粘连程度进行评分;HE 染色验证子宫内膜异位症模型是否成功;Masson 染色检测异位灶胶原纤 维沉积情况,评估其胶原沉积程度;免疫组化法检测纤维化相关蛋白 E-cadherin、Collagen I、α-SMA、smooth muscle myosin heavy chain II (SMMHC-II) 的表达情况,评估异位灶纤维化程度。 结果 改良腹腔注射法构建小鼠子宫内膜异位症纤维化小鼠模型成功率可达到 100%。 小鼠腹腔灌洗液 TGF-β1 水平随造模时间延长有明显升高。 Masson 染色和纤维化相关蛋白免疫组化染色观察到子宫内膜异位症小鼠腹腔存在纤维化表型。 结论 改良的腹 腔注射法构建小鼠子宫内膜异位症纤维化模型成功率高,腹腔灌洗液 TGF-β1 水平、Masson 染色和纤维化相关蛋白 E-cadherin、Collagen I、α-SMA、SMMHC-II 免疫组化染色能够评价小鼠子宫内膜异位症盆腔纤维化程度。

    Abstract:

    Objective To establish a mouse model of endometriosis with fibrosis and to study its application. Methods A mouse model of endometriosis with fibrosis was established by modified intraperitoneal injection using BALB/ c mice; the peritoneal lavage fluid of mice was collected and the level of TGF-β1 in lavage fluid was detected by ELISA. The degree of abdominal adhesion was scored and the success of the endometriosis model was verified by hematoxylin-eosin(HE)staining, while the collagen deposition in the ectopic focus was detected by Masson staining and the degree of collagen deposition was evaluated. The expression of fibrosis-related proteins E-cadherin, collagen I, α-SMA, and smooth muscle myosin heavy chain II ( SMMHC-II) were detected by immunohistochemistry, and the degree of ectopic fibrosis was evaluated. Results The success rate of a mouse model of endometriosis with fibrosis constructed by a modified intraperitoneal injection method could reach 100%. The level of TGF-β1 in the peritoneal lavage fluid of mice increased significantly with the extension of modeling time. Masson staining and immunohistochemical staining of fibrosis-related proteins revealed a fibrotic phenotype in the abdominal cavity of endometriosis mice. Conclusions The improved intraperitoneal injection method has a high success rate in the establishment of a mouse model of endometriosis with fibrosis. TGF-β1 levels in peritoneal lavage fluid, Masson staining, and immunohistochemical staining of fibrosis-related proteins can evaluate the degree of pelvic fibrosis in mice with endometriosis.

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毕艳丽,倪喆鑫,李溪,俞超芹.子宫内膜异位症纤维化小鼠模型构建方法及其应用[J].中国实验动物学报,2020,28(4):447~454.

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  • 收稿日期:2020-02-01
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  • 在线发布日期: 2020-09-15
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