吉林大学学报(医学版) ›› 2023, Vol. 49 ›› Issue (1): 215-221.doi: 10.13481/j.1671-587X.20230129

• 方法学 • 上一篇    

化学和热传导双重损伤构建大鼠宫腔黏连模型的方法及其评价

笪琳萃,郑备红,孙艳,陈素珠,林运鸿,陈清汾,杜生荣()   

  1. 福建省妇幼保健院 福建医科大学妇儿临床医学院生殖医学中心,福建 福州 350001
  • 收稿日期:2022-03-17 出版日期:2023-01-28 发布日期:2023-02-03
  • 通讯作者: 杜生荣 E-mail:dushengrong2001@126.com
  • 作者简介:笪琳萃(1989-),女,江西省德兴市人,助理研究员,理学博士,主要从事再生医学相关方面的研究。
  • 基金资助:
    福建省卫健委中青年骨干人才培养资助项目(2019-ZQNB-16);福建省科技厅自然科学基金项目(2019J01511);福建省妇幼保健院科技创新启动基金项目(妇幼YCXB 18-03)

Method to establish rat model of intrauterine adhesion through chemical and heat conduction dual injury and its evaluation

Lincui DA,Beihong ZHENG,Yan SUN,Suzhu CHEN,Yunhong LIN,Qingfen CHEN,Shengrong DU()   

  1. Reproductive Center,Fujian Provincial Maternity and Children’s Hospital,College of Clinical Medicine for Obstetrics & Gynecology and Pediatrics,Fujian Medical University,Fuzhou 350001,China
  • Received:2022-03-17 Online:2023-01-28 Published:2023-02-03
  • Contact: Shengrong DU E-mail:dushengrong2001@126.com

摘要:

目的 采用化学和热传导双重损伤法构建大鼠宫腔黏连(IUA)模型,阐明该方法作为化学损伤的改良方法构建接近 IUA 病理学变化动物模型的特点和优势。 方法 6 只 6~8 周龄雌性 SD 大鼠随机分为化学损伤组和化学及热传导双重损伤组(双重损伤组),每组3只。以大鼠左侧宫角为实验组,右侧宫角为自身对照组。化学损伤组大鼠左侧宫角作为化学损伤实验组采用95% 乙醇作用3 min,右侧宫角作为化学损伤自身对照组不进行任何处理;双重损伤组大鼠左侧宫角作为双重损伤实验组采用95% 乙醇作用 3 min 后100 ℃ 热水再处理1 min,右侧宫角作为双重损伤自身对照组不进行任何处理。造模术后第14天,采用 HE染色、Masson 染色和 Ki67 免疫组织化学染色法检测各组大鼠子宫内膜厚度、腺体数量、纤维化面积比及腺体上皮细胞和基质细胞的增殖指数(PI)。 结果 HE 染色,与相对应的自身对照组比较,化学损伤实验组和双重损伤实验组大鼠子宫内膜厚度和腺体数量明显减少(P<0.05);与化学损伤实验组比较,双重损伤实验组大鼠子宫宫腔封闭,子宫内膜厚度和腺体数量均明显减少(P<0.05)。Masson 染色,与相对应的自身对照组比较,化学损伤实验组和双重损伤实验组大鼠子宫内膜层呈纤维化增生,子宫内膜组织中纤维化面积比明显升高(P<0.05);与化学损伤实验组比较,双重损伤实验组大鼠子宫内膜层可见纤维化黏连带和完整肌层,子宫内膜组织中纤维化面积比明显升高(P<0.05)。Ki67 免疫组织化学染色,与相对应的自身对照组比较,化学损伤实验组和双重损伤实验组大鼠子宫内膜组织中腺体上皮细胞核形状和排列发生改变,基质细胞 PI 明显升高(P<0.05)。 结论 化学和热传导双重损伤法操作简便且可重复性好,可为 IUA 治疗的安全性评价、机制研究和临床转化提供具有纤维化黏连带和完整肌层的IUA模型,具有良好的应用前景。

关键词: 宫腔黏连, 动物模型, 子宫内膜, 化学损伤法, 热传导损伤法

Abstract:

Objective To establish the rat models of intrauterine adhesion (IUA) with the chemical and heat conduction dual injury method, and to clarify its characteristics and advantages as an improved approach of chemical injury in constructing the animal model with similar pathological changes of IUA. Methods Six female SD rats aged 6-8 weeks were randomly divided into chemical injury group and chemical and heat conduction dual injury group (dual injury group),with 3 rats in each group.The left uterine horns of rats were used as experimental group and the right uterine horns were used as self-control group. The left uterine horns of the rats in chemical injury group, which were defined as the chemical injury experiment group, were treated with 95% ethanol for 3 min; the right uterine horns of the rats, which were defined as chemical injury self-control group,didn’t received any treatment. The left uterine horns of the rats in dual injury group, which were defined as dual injury experiment group, were treated with 95% ethanol in the left uterus for 3 min, and then treated with 100 ℃ hot water for 1 min; the right uterine horns of the rats, which were defined as dual injury self-control group, didn’t receive any treatment.On the 14 th day after modeling, the thickness of endometrial, the number of glands, the area of endometrial fibrosis and the proliferation index (PI) of glandular epithelial cells and stromal cells of the rats in various groups were detected by hematoxylin and eosin (HE) staining, Masson staining, and Ki67 immunohistochemical staining method. Results The HE staining results showed that compared with self-control group,the thickness of endometrium of the rats and the number of glands in chemical injury experiment group and dual injury experiment group were significantly reduced(P<0.05).Compared with chemical injury experiment group, the uterine cavity of the rats in dual injury experiment group was closed, the endometrial thickness was reduced (P<0.05), and the number of endometrial glands was decreased(P<0.05). The Masson staining results showed that compared with self-control group, the fibrotic hyperplasia in endometrial layers of the rats in chemical injury experiment group and dual injury experiment group was seen, and the ratios of endometrial fibrosis area in exdometrium tissue were apparently increased (P<0.05). Compared with chemical injury experiment group, fibrotic adhesion and intact myometrium of the rats in dual injury experiment group were found, and the ratio of endometrial fibrosis area was apparently increased (P<0.05). The Ki67 immunohistochemical staining results showed that compared with self-control group, the nuclear shape and arrangement of glandular epithelial cells appeared the changes in endometrium tissue of the rats in chemical injury experiment group and dual injury experiment group, and the PI was significantly increased (P<0.05). Conclusion Chemical and heat conduction dual injury method is an easy and reproducible operation that can establish the IUA model with fibrotic adhesion and intact muscle layer for the safety evaluation, mechanism research and clinical transformation of IUA, which possesses a good application perspect.

Key words: Intrauterine adhesion, Animal model, Endometrium, Chemical injury method, Heat conduction injury method

中图分类号: 

  • R711.74