| 常品,张胜辉,陈童彤,孙钰椋,张花,潘莹,刘彦礼,林俊堂.经宫颈机械损伤联合脂多糖灌注法构建小鼠宫腔黏连模型[J].中国实验动物学报,2022,30(3):400~407. |
| 经宫颈机械损伤联合脂多糖灌注法构建小鼠宫腔黏连模型 |
| Establishment of a mouse intrauterine adhesion model by transcervical mechanical injury combined with lipopolysaccharide perfusion |
| 投稿时间：2021-10-25 |
| DOI：10. 3969 / j.issn.1005-4847. 2022. 03. 013 |
| 中文关键词: 宫腔粘连 机械损伤 脂多糖 动物模型 |
| 英文关键词:intrauterine adhesion mechanical injury lipopolysaccharide animal model |
| 基金项目: |
| 作者 | 单位 | E-mail | | 常品 | 1. 新乡医学院第三附属医院,河南 新乡 453003
2. 新乡医学院干细胞与生物治疗技术研究中心,河南 新乡 453003 | pin282669919@ 163. com | | 张胜辉 | 1. 新乡医学院第三附属医院,河南 新乡 453003
2. 新乡医学院干细胞与生物治疗技术研究中心,河南 新乡 453003 | | | 陈童彤 | 2. 新乡医学院干细胞与生物治疗技术研究中心,河南 新乡 453003
3. 新乡医学院生命科学技术学院,河南 新乡 453003 | | | 孙钰椋 | 2. 新乡医学院干细胞与生物治疗技术研究中心,河南 新乡 453003
4. 新乡医学院医学工程学院,河南 新乡 453003 | | | 张花 | 新乡医学院第三附属医院,河南 新乡 453003 | | | 潘莹 | 新乡医学院第三附属医院,河南 新乡 453003 | panying@ xxmu. edu. cn | | 刘彦礼 | 2. 新乡医学院干细胞与生物治疗技术研究中心,河南 新乡 453003
3. 新乡医学院生命科学技术学院,河南 新乡 453003 | | | 林俊堂 | 2. 新乡医学院干细胞与生物治疗技术研究中心,河南 新乡 453003
4. 新乡医学院医学工程学院,河南 新乡 453003 | |
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| Author Name | Affiliation | E-mail | | CHANG Pin | 1.the Third Affiliated Hospital of Xinxiang Medical University, Xinxiang 453003, China. 2. Stem Cell and Biotherapy Technology Research Center, Xinxiang 453003. | pin282669919@ 163. com | | ZHANG Shenghui | 1.the Third Affiliated Hospital of Xinxiang Medical University, Xinxiang 453003, China. 2. Stem Cell and Biotherapy Technology Research Center, Xinxiang 453003. | | | CHEN Tongtong | 2. Stem Cell and Biotherapy Technology Research Center, Xinxiang 453003. 3. College of Life Science and Technology, Xinxiang Medical University, Xinxiang 453003. | | | SUN Yuliang | 2. Stem Cell and Biotherapy Technology Research Center, Xinxiang 453003.4. College of Medical Engineering, Xinxiang Medical University, Xinxiang 453003 | | | ZHANG Hua | the Third Affiliated Hospital of Xinxiang Medical University, Xinxiang 453003, China | | | PAN Ying | the Third Affiliated Hospital of Xinxiang Medical University, Xinxiang 453003, China | panying@ xxmu. edu. cn | | LIU Yanli | 2. Stem Cell and Biotherapy Technology Research Center, Xinxiang 453003. 3. College of Life Science and Technology, Xinxiang Medical University, Xinxiang 453003. | | | LIN Juntang | 2. Stem Cell and Biotherapy Technology Research Center, Xinxiang 453003.4. College of Medical Engineering, Xinxiang Medical University, Xinxiang 453003 | |
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| 中文摘要: |
| 目的 通过微创的方式,利用机械损伤联合脂多糖灌注法构建符合临床损伤特征的宫腔黏连 (intrauterine adhesion,IUA)动物模型,为深入研究 IUA 的发病机制、病理变化及探索临床治疗方案提供支持。 方法 取雌性 ICR 小鼠 20 只随机分为 2 组(n= 10),间苯三酚松弛宫颈后,造模组动物经宫颈给予宫腔内机械损伤+脂多糖(LPS)灌注(0. 5 mg / kg),假手术组动物仅接受等体积生理盐水灌注。 分别于术后 7、14、21 d 收集子宫标本, 常规 HE 和 Masson 染色,检测子宫内膜形态及纤维化情况;免疫荧光检测子宫内膜中巨噬细胞(F4 / 80)分布,qPCR 及 Western Blot 检测造模 14 d 小鼠子宫内膜中炎症因子(IL-1β、TNF-α、IL-4、IL-10 和 IL-6)及子宫内膜容受性相关标志物(Integrin β3 和 LIF)的表达变化。 随后,取 20 只小鼠随机分为经宫颈机械损伤+脂多糖灌注组(n= 10)为造模组和假手术组(n= 10),造模 2 周后合笼,记录妊娠母鼠数量;妊娠约 18 d 处死母鼠,记录胚胎数量。 结果 机械损伤+脂多糖灌注后小鼠子宫中炎症反应和纤维化程度显著增加。 其中模型组小鼠子宫内膜厚度较假手术组显著降低,子宫内膜细胞坏死明显,腺体缺失,腔上皮和腺上皮细胞结构不完整,组织纤维化严重。 随后,检测发现巨噬细胞标志物 F4 / 80 在模型组小鼠子宫内膜中表达显著上升(P< 0.05),并且模型组小鼠子宫内膜中促炎因子 IL-1β、TNF-α、和 IL-6 表达显著升高(P< 0.05),而抑炎因子 IL-4、IL-10 表达明显下降(P< 0.05)。 进一步研究发现子宫内膜容受性标志物 LIF 和 Integrin β3 表达量显著下降(P< 0.05),且生育实验结果发现妊娠母鼠及胎鼠数量均显著下降(P< 0.05)。 结论 本研究通过微创的方式经宫颈机械损伤+脂多糖灌注,成功建立小鼠 IUA 模型构建方法,符合临床 IUA 损伤特征,可为后续针对 IUA 发病机制及治疗方法研究提供稳定的动物模型。 |
| 英文摘要: |
| Objective To establish a stable animal model of intrauterine adhesion ( IUA) using a minimally invasive method that recapitulates the clinicopathologic characteristics of IUA. Methods Overall, 20 female ICR mice were randomly divided into two groups (n= 10). After the cervix was relaxed with phloroglucinol, the uterine horn of mice in the model group was subjected to mechanical injury and lipopolysaccharide perfusion (0. 5 mg / kg) through the cervix to induce endometrial injury, while mice in the sham operation group only received the same volume of normal saline perfusion. Uteri of the mice were collected at 7, 14 and 21 days after the procedure. HE and Masson staining were used to assess uterine morphology and fibrosis. Immunofluorescence was performed to evaluate macrophage infiltration into the endometrium. qPCR and Western Blot were used to detect the expression of inflammatory factors ( IL-1β, TNF-α, IL-4, IL-10 and IL-6) and endometrial receptivity-related markers ( integrin β3 and LIF) in mice at 14 days after modeling. Furthermore, mouse fertility was evaluated in model and sham operation groups. Results Compared with the sham operation group, endometria in the model group were significantly thinner and exhibited severe necrosis, glandular loss, and incomplete luminal epithelial and glandular epithelial cell structures. Subsequently, severe tissue fibrosis was observed in endometria in the model group. Additionally, increased macrophage infiltration and upregulated expression of proinflammatory factors demonstrated an activated inflammatory response in endometria of the model group. Moreover, endometrial receptivity was significantly decreased after mechanical injury combined with lipopolysaccharide perfusion. Finally, significant decreases in pregnancy and the number of fetuses were observed in the model group. Conclusions This study demonstrated successful establishment of an IUA mouse model through a minimally invasive transcervical mechanical injury combined with lipopolysaccharide perfusion, which was consistent with the clinical characteristics of severe IUA. This model might support in-depth study of IUA pathogenesis and promote the development of IUA therapies. |
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