间充质干细胞修复大鼠子宫切口瘢痕缺陷的实验研究

doi:10.3760/cma.j.issn.1007-1245.2024.14.012

摘要/Abstract

摘要：

目的　探讨间充质干细胞（MSCs）对大鼠子宫切口瘢痕缺陷（PCSD）的修复效果，并为瘢痕妊娠的治疗提供潜在策略。方法　实验在广东省医学实验动物中心进行，实验时间为2023年12月至2024年1月。选取雌性特定病原体自由级Sprague-Dawley（SPF级SD）大鼠120只，7～8周龄，体质量为300～350 g，按照随机数字表法分为3组，每组40只。通过连续两次的剖宫产建立大鼠子宫憩室模型并通过B超和组织学验证。对照组接受0.2 ml的局部生理盐水注射和0.5 ml的尾静脉生理盐水注射；实验组-点位注射组局部注射0.2 ml的MSCs凝胶；实验组-尾静脉注射组通过尾静脉注射0.2 ml MSCs凝胶。对3组子宫切口瘢痕的修复效果进行观察，系统记录大鼠的饮食、饮水和精神状态表现。给药结束后第4周处死并解剖大鼠，对其脏器进行观察。进一步取子宫组织样本进行苏木精-伊红（HE）染色，观测子宫肌层和内膜的厚度，并计算子宫内膜缺陷率、α-平滑肌肌动蛋白（α-SMA）和转化生长因子-β1（TGF-β1）阳性面积百分比。统计学方法采用t检验、方差分析和χ²检验。结果　实验组中正常饮食、饮水、精神状态正常的大鼠比例高于对照组［72.5%（58/80）比52.5%（21/40）、76.3%（61/80）比42.5%（17/40）、70.0%（56/80）比47.5%（19/40）］，差异均有统计学意义（χ²=4.74、13.35、6.46，均P<0.05）。HE染色结果显示，点位注射组大鼠子宫内膜腺体增生、少量纤维组织增生、炎症细胞浸润，肌层见巨噬细胞、手术缝线。尾静脉注射组大鼠子宫内膜腺体增生、少量纤维组织增生、炎症细胞浸润，肌层见巨噬细胞、手术缝线。尾静脉注射组与点位注射组的基层厚度、内膜厚度、α-SMA阳性面积百分比均高于对照组［（116.32±4.59）μm比（109.21±5.73）μm比（74.23±6.30）μm、（200.34±7.21）μm比（156.98±6.98）μm比（119.55±7.38）μm、（32.99±1.26）%比（25.93±1.33）%比（20.59±1.48）%］，TGF-β1阳性面积百分比、子宫内膜缺陷均低于对照组［（14.11±1.01）%比（17.54±1.08）%比（20.09±1.13）%、35.00%（14/40）比62.50%（25/40）比100.00%（40/40）］，差异均有统计学意义（F=650.86、1 264.19、836.93、311.99，χ²=37.86，均P<0.05）。结论　脐带MSCs能促进子宫憩室的修复，具有良好的安全性，有望为子宫切口后瘢痕形成的治疗提供新的策略。

关键词:

间充质干细胞, 大鼠造模, 子宫切口瘢痕缺陷, 动物实验

Abstract:

Objective　To investigate the repair effect of mesenchymal stem cells (MSCs) on rat previous cesarean scar defect (PCSD) and to provide a potential strategy for the treatment of scar pregnancy. Methods　This research experiment was conducted from December 2023 to January 2024 at Guangdong Medical Laboratory Animal Center. A total of 120 female specific pathogen free grade Sprague-Dawley (SPF SD) rats, aged 7 to 8 weeks and weighing 300 to 350 g, were selected and were divided into 3 groups with 40 rats per group according to the random number table method. A rat uterine diverticulum model was established through two consecutive cesarean sections and verified by ultrasound and histology. The control group received 0.2 ml of local saline injection and 0.5 ml of tail vein saline injection, the local injection group (experimental group) was injected with 0.2 ml of MSCs gel locally, and the tail vein injection group (experimental group) was injected with 0.2 ml of MSCs gel through the tail vein. The repair effects on uterine scar defects were observed. The rats' diet, water intake, and mental status were systematically recorded. Four weeks post-administration, the rats were euthanized for autopsy, and their organs were macroscopically observed. Uterine tissue samples were collected for hematoxylin and eosin (H&E) staining to observe the thickness of the myometrium and endometrium, and the endometrial defect rate, the percentage of α-smooth muscle actin (α-SMA) positive area, and the percentage of transforming growth factor-β1 (TGF-β1) positive area were calculated. The statistical methods used were t test, ANOVA, and χ² test. Results　The proportions of the rats with normal diet, water intake, and mental status in the experimental group were higher than those in the control group [72.5% (58/80) vs. 52.5% (21/40), 76.3% (61/80) vs. 42.5% (17/40), 70.0% (56/80) vs. 47.5% (19/40)], with statistically significant differences (χ²=4.74, 13.35, and 6.46, all P<0.05). H&E staining revealed that in both local and tail vein injection groups, there was an increase in endometrial gland proliferation, a small amount of fibrous tissue proliferation, and inflammatory cell infiltration, with macrophages and surgical sutures observed in the muscle layer. The basal layer thickness, endometrial thickness, and percentage of α-SMA positive area in the tail vein and local injection groups were higher than those in the control group [(116.32±4.59) μm vs. (109.21±5.73) μm vs. (74.23±6.30) μm, (200.34±7.21) μm vs. (156.98±6.98) μm vs. (119.55±7.38) μm, (32.99±1.26)% vs. (25.93±1.33)% vs. (20.59±1.48)%], but the percentage of TGF-β1 positive area and endometrial defect rate were lower than those in the control group [(14.11±1.01)% vs. (17.54±1.08)% vs. (20.09±1.13)%, 35.00% (14/40) vs. 62.50% (25/40) vs. 100.00% (40/40)], with statistically significant differences (F=650.86, 1 264.19, 836.93, and 311.99, χ²=37.86, all P<0.05). Conclusions　Umbilical cord MSCs can significantly promote the repair of uterine diverticula and are safe. This provides an experimental basis for the application of MSCs in the treatment of uterine scar pregnancy and offers a new strategy for the treatment of post-cesarean scar formation.

Key words:

Mesenchymal stem cells, Rat modeling, Previous cesarean scar defect, Animal experiment

袁雪青朱梅.

间充质干细胞修复大鼠子宫切口瘢痕缺陷的实验研究 [J]. 国际医药卫生导报, 2024, 30(14): 2343-2347.

Yuan Xueqing, Zhu Mei.

Experimental study of mesenchymal stem cell repair of previous cesarean scar defect in rats [J]. International Medicine and Health Guidance News, 2024, 30(14): 2343-2347.

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