多重方法建立大鼠子宫内膜深度损伤模型

张奕; 胡秀娟; 戴志俊; 刘超; 洪名云

doi:10.16766/j.cnki.issn.1674-4152.001948

多重方法建立大鼠子宫内膜深度损伤模型

doi: 10.16766/j.cnki.issn.1674-4152.001948

张奕^{1, 2},
胡秀娟³,
戴志俊^{1, 2},
刘超³,
洪名云^{1, 2, ,}

1.
安徽省妇幼保健院生殖中心，安徽合肥 230001
2.
安徽医科大学附属妇幼保健院生殖中心，安徽合肥 230001
3.
安徽医科大学组织胚胎学教研室，安徽合肥 230032

基金项目:

安徽省重点研究与开发计划项目 1704a0802171

合肥市“借转补”基金项目 J2018Y03

合肥市第六周期医学重点学科建设专科合卫科教[2019]160号

详细信息

通讯作者:
洪名云，E-mail：2016mingyun@sina.com

中图分类号: R711.74 R-332
计量
- 文章访问数: 258
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- PDF下载量: 16
- 被引次数: 0
出版历程
- 收稿日期: 2020-08-05
- 网络出版日期: 2022-02-16

Multiple methods to establish rat endometrial deep injury model

ZHANG Yi^{1, 2},
HU Xiu-juan³,
DAI Zhi-jun^{1, 2},
LIU Chao³,
HONG Ming-yun^{1, 2
, ,}

1.
Reproductive Center of Anhui Maternal and Child Health Hospital, Hefei, Anhui 230001, China

摘要

摘要: 目的建立一种大鼠子宫内膜深度损伤的动物模型，为子宫内膜干细胞复合生物支架疗法提供动物模型。方法将15只健康有正常动情周期的雌性大鼠按照随机数表的分组方式分为假手术组(5只)及模型组(10只)；假手术组雌性大鼠仅开腹不对子宫进行操作，模型组雌性大鼠开腹后对每只大鼠左侧子宫做手术切口后行子宫内膜机械搔刮及宫腔注入细菌脂多糖溶液、子宫缺血处理，右侧子宫做手术切口但不做其他任何处理。经过2个动情周期，假手术组及模型组大鼠均于动情期开腹收集子宫组织，行HE和Masson染色，免疫组织化学检测内膜Collagen Ⅰ、CK18、Vimentin及PECAM-1的表达，对损伤子宫内膜的腺体数目和纤维化面积比率，内膜Collagen Ⅰ、CK18、Vimentin及PECAM-1的表达水平进行统计学分析。结果病理学结果显示模型组子宫内膜明显变薄，腺体数量减少，结构紊乱，间质纤维组织增多，胶原聚集，纤维化较明显。免疫组织化学结果显示模型组子宫内膜Collagen Ⅰ表达水平比假手术组高，CK18、Vimentin、PECAM-1表达水平低于假手术组，差异有统计学意义(均P < 0.05)。结论采用机械损伤、宫腔注入细菌脂多糖及缺血三重损伤法可以建立大鼠子宫内膜深度损伤模型。
- 子宫内膜 /
- 损伤 /
- SD大鼠
Abstract: Objective To establish an animal model of rat endometrial deep injury, and to provide a basis for subsequent treatment of endometrial injury with endometrial stem cells combined with biological scaffolds. Methods Fifteen healthy female rats with normal oestrous cycle were randomly divided into the following two groups: 5 rats in the sham operation group and 10 rats in the model group. The female rats in the sham operation group only performed laparotomy without uterus operation. After laparotomy, the left uterus of each rat in the model group received mechanical endometrial scratching, injection of lipopolysaccharide solution and uterine ischemia. The right uterus received surgical incision without any other treatment. After two oestrus cycles, oestrus laparotomy uterine tissues were collected from control group and model group rats. HE and Masson staining and immunohistochemical detection were used for the lining of Collagen Ⅰ; the expression of CK18, Vimentin, PECAM-1 and the damage area ratio of the number of endometrial glands and fibrosis. The expression levels of lining collagen Ⅰ, CK18, Vimentin and PECAM-1 were used for statistical analysis. Results Pathological observation showed that the endometrium in the model group was significantly thinner, the number of glands decreased, the structure was disordered, the interstitial fibrous tissue increased and collagen aggregation and fibrosis were obvious. In the immunohistochemical results, the expression level of Collagen Ⅰ in the model group was higher than that in the sham group, whereas the expression levels of CK18, Vimentin and PECAM-1 were lower than that in the sham group, with statistically significant differences (all P < 0.05). Conclusion The deep injury model of rat endometrium can be established by mechanical injury, bacterial lipopolysaccharide injection and triple injury after ischemia.
- Endometrium /
- Damage /
- SD rats

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图 1 模型组大鼠子宫内膜HE染色(×40)

注：模型组大鼠内膜腺体数量减少。

下载: 全尺寸图片幻灯片

图 2 模型组大鼠子宫内膜Masson染色(×40)

注：模型组大鼠子宫内膜纤维化面积增大。

下载: 全尺寸图片幻灯片

图 3 模型组大鼠子宫内膜Collagen Ⅰ的表达(×40)

注：免疫组化染色，模型组大鼠子宫内膜基质有大量Ⅰ型胶原蛋白表达。

下载: 全尺寸图片幻灯片

图 4 模型组大鼠子宫内膜ck18表达(×40)

注：免疫组化染色，模型组子宫内膜只有部分腺体残存的CK18, 大多数腺细胞呈阴性。

下载: 全尺寸图片幻灯片

图 5 模型组大鼠子宫内膜Vimentin表达水平(×40)

注：免疫组化染色，模型组子宫内膜Vimentin表达降低。

下载: 全尺寸图片幻灯片

图 6 模型组大鼠子宫内膜PECAM-1的表达(×40)

注：免疫组化染色，模型大鼠子宫内膜PECAM-1表达降低

下载: 全尺寸图片幻灯片

参考文献(15)

[1]	HANSON B, JOHNSTONE E, DORAIS J, et al. Female infertility, infertility-associated diagnoses, and comorbidities: a review[J]. J Assist Reprod Genet, 2017, 34(2): 167-177. doi: 10.1007/s10815-016-0836-8
[2]	宋天然, 孙海翔. 子宫内膜损伤致不孕症的临床处理[J]. 中国实用妇科与产科杂志, 2020, 36(6): 500-504. https://www.cnki.com.cn/Article/CJFDTOTAL-ZGSF202006006.htm
[3]	LEE Y J, YI K W. Bone marrow-derived stem cells contribute to regeneration of the endometrium[J]. Clin Exp Reprod Med, 2018, 45(4): 149-153. doi: 10.5653/cerm.2018.45.4.149
[4]	BOSTEELS J, WEYERS S, D'HOOGHE T M, et al. Anti-adhesion therapy following operative hysteroscopy for treatment of female subfertility[J]. Cochrane Database Syst Rev, 2017, 11(11): pCD011110. http://www.researchgate.net/profile/Jan_Bosteels/publication/321317080_Anti-adhesion_therapy_following_operative_hysteroscopy_for_treatment_of_female_subfertility/links/5ac10166aca27222c75b4d51/Anti-adhesion-therapy-following-operative-hysteroscopy-for-treatment-of-female-subfertility.pdf
[5]	刘彩姣, 王蔼明. 干细胞在子宫内膜损伤修复中的研究进展[J]. 安徽医药, 2017, 21(5): 781-785. doi: 10.3969/j.issn.1009-6469.2017.05.001
[6]	TERSOGLIO A E, TERSOGLIO S, SALATINO D R, et al. Regenerative therapy by endometrial mesenchymal stem cells in thin endometrium with repeated implantation failure. A novel strategy[J]. JBRA Assist Reprod, 2020, 24(2): 118-127. http://www.researchgate.net/publication/333115282_O-08_Regenerative_therapy_by_endometrial_mesenchymal_stem_cells_in_thin_endometrium_with_repeated_implantation_failure_A_novel_strategy/download
[7]	吴瑾柔, 骆婕, 陶莹. 干细胞治疗子宫内膜损伤及促进生育的研究进展[J]. 重庆医学, 2019, 48(16): 2843-2846. doi: 10.3969/j.issn.1671-8348.2019.16.035
[8]	王萌, 刘晨, 夏波. 宫腔粘连的诊疗进展[J]. 医学信息, 2020, 33(8): 35-39. doi: 10.3969/j.issn.1006-1959.2020.08.013
[9]	南楠, 梁敏, 刘婷婷, 等. 乙醇作用时间对大鼠子宫内膜损伤的影响[J]. 生殖医学杂志, 2019, 28(1): 66-70. doi: 10.3969/j.issn.1004-3845.2019.01.013
[10]	许春燕, 宋阳, 李坤寅, 等. 薄型子宫内膜模型的动物选择及其造模方法改进[J]. 中国实验动物学报, 2016, 24(2): 217-220. doi: 10.3969/j.issn.1005-4847.2016.02.021
[11]	张引红, 王春芳, 陈朝阳, 等. 薄型子宫内膜小鼠模型的建立及初步鉴定[J]. 山西医科大学学报, 2017, 48(5): 432-435. https://www.cnki.com.cn/Article/CJFDTOTAL-SXYX201705006.htm
[12]	韩笑, 黄晓武. 干细胞在子宫内膜损伤后修复的研究进展[J]. 国际妇产科学杂志, 2019, 46(4): 365-369. doi: 10.3969/j.issn.1674-1870.2019.04.001
[13]	蔡慧华, 何援利, 李慧娟, 等. 宫腔粘连大鼠模型的建立及其与白细胞介素21的相关性分析[J]. 医学研究生学报, 2015, 28(4): 346-349. doi: 10.3969/j.issn.1008-8199.2015.04.003
[14]	冯妍, 倪和民, 郭勇, 等. 脂多糖对体外培养奶牛子宫内膜上皮细胞子宫容受性因子表达的影响[J]. 中国畜牧兽医, 2019, 46(8): 2288-2295. https://www.cnki.com.cn/Article/CJFDTOTAL-GWXK201908013.htm
[15]	孙瑶琦, 蒋惠慈, 刘洁. 炎症因子与子宫腺肌病发病机制研究进展[J]. 国际妇产科学杂志, 2020, 47(1): 96-100. doi: 10.3969/j.issn.1674-1870.2020.01.023