[脐带血间充质干细胞对兔宫内粘连模型的影响]。

Q3 Medicine
J J Wang, Q Hua, H J Li, D M Zhang, Y L Wang
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Four weeks after the IUA modeling surgery, the rabbits were euthanized and samples were collected. Hematoxylin and eosin (HE) staining and Masson staining were used to assess the number of endometrial glands and the fibrosis rate. RNA sequencing was performed on the endometrium of the IUA group and hUCB-MSC transplantation group.The mRNA and protein expression of the fibrosis markers [transforming growth factor β1 (TGF-β1) and tissue inhibitors of metalloproteinase 1 (TIMP1)] and RNA sequencing-related parameters were detected by quantitative real-time PCR (qRT-PCR) and Western blot. <b>Results:</b> Compared with the control group, the number of glands in IUA group decreased (26.33±1.53 vs 4.33±1.53, <i>P</i><0.001), and the fibrosis rate increased (18.01%±2.21% vs 69.55%±2.42%, <i>P</i><0.001), indicating successful modeling. 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引用次数: 0

摘要

目的通过建立动物模型,研究人脐血间充质干细胞(hUCB-MSC)对宫腔内粘连(IUA)子宫内膜的修复作用。研究方法按随机数字表法将18只健康雌性新西兰白兔分为对照组、IUA组和hUCB-间充质干细胞移植组。对照组仅进行开腹手术。IUA 组进行 IUA 造模手术,采用刮宫术,并在子宫腔内放置脂多糖(LPS)棉花进行双重损伤。hUCB-间充质干细胞移植组接受同样的IUA建模手术,术后一周在双侧子宫肌层多点注射hUCB-间充质干细胞悬液(2×106个细胞/毫升,500微升)。IUA 造模手术四周后,兔子被安乐死并采集样本。采用苏木精和伊红(HE)染色法和马森染色法评估子宫内膜腺体的数量和纤维化率。通过实时定量PCR(qRT-PCR)和Western blot检测纤维化标志物[转化生长因子β1(TGF-β1)和金属蛋白酶1组织抑制剂(TIMP1)]的mRNA和蛋白表达以及RNA测序相关参数。结果与对照组相比,IUA 组腺体数量减少(26.33±1.53 vs 4.33±1.53,PPPP=0.001)。纤维化相关蛋白(TGF-β1和TIMP1)的Western印迹分析显示,与对照组相比,IUA组TGF-β1(0.91±0.05)和TIMP1(0.99±0.01)蛋白水平升高(0.61±0.04,0.68±0.07)(P=0.001,0.015)。与 IUA 组相比,hUCB-间充质干细胞移植组的 TGF-β1 (0.69±0.04)和 TIMP1 (0.62±0.08)蛋白表达量减少(P=0.005,0.014)。与PI3K/AKT信号通路相关的Western Blot结果显示,IUA组p-PI3K(1.05±0.05)和p-AKT(1.17±0.06)的表达量较对照组(0.78±0.03,0.85±0.05)有所增加(P=0.002,0.002)。与 IUA 组相比,hUCB-MSC 移植组 p-PI3K (0.74±0.02)和 p-AKT(0.93±0.04)蛋白的表达下降(P=0.003,0.005)。结论:hUCB-间充质干细胞可通过增加子宫内膜腺体数量和改善其纤维化水平来修复兔宫腔内粘连模型中受损的子宫内膜。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
[Effect of mesenchymal stem cell derived from umbilical cord blood on rabbit intrauterine adhesion model].

Objective: To investigate the repair effect of human umbilical cord blood mesenchymal stem cells (hUCB-MSC) on endometrium of intrauterine adhesion (IUA) by establishing animal model. Methods: Eighteen healthy female New Zealand white rabbits were divided into a control group, an IUA group and an hUCB-MSC transplantation group according to random number table method. The control group underwent laparotomy only. The IUA group underwent IUA modeling surgery using curettage and lipopolysaccharide (LPS) cotton placed in uterine cavity for double injury. The hUCB-MSC transplantation group received the same IUA modeling surgery followed by multipoint injection of hUCB-MSC suspension (2×106 cells/ml, 500 μl) into the bilateral uterine myometrium one week after surgery. Four weeks after the IUA modeling surgery, the rabbits were euthanized and samples were collected. Hematoxylin and eosin (HE) staining and Masson staining were used to assess the number of endometrial glands and the fibrosis rate. RNA sequencing was performed on the endometrium of the IUA group and hUCB-MSC transplantation group.The mRNA and protein expression of the fibrosis markers [transforming growth factor β1 (TGF-β1) and tissue inhibitors of metalloproteinase 1 (TIMP1)] and RNA sequencing-related parameters were detected by quantitative real-time PCR (qRT-PCR) and Western blot. Results: Compared with the control group, the number of glands in IUA group decreased (26.33±1.53 vs 4.33±1.53, P<0.001), and the fibrosis rate increased (18.01%±2.21% vs 69.55%±2.42%, P<0.001), indicating successful modeling. HE and Masson staining revealed that the number of glands in the hUCB-MSC transplantation group was increased compared with that in IUA group (17.33±2.52 vs 4.33±1.53, P<0.001), and the fibrosis rate decreased (69.55%±2.42% vs 41.55%±1.99%,P=0.001). Western blot analyses of fibrosis-related proteins (TGF-β1 and TIMP1) showed elevated levels of TGF-β1 (0.91±0.05) and TIMP1 (0.99±0.01) proteins in the IUA group compared to control group (0.61±0.04, 0.68±0.07) (P=0.001, 0.015). The hUCB-MSC transplantation group exhibited reduced expression of TGF-β1 (0.69±0.04) and TIMP1 (0.62±0.08) proteins compared to the IUA group (P=0.005, 0.014). Western Blot results related to the PI3K/AKT signaling pathway [phosphorylated phosphatidylinositol 3-kinase (p-PI3K), phosphorylated protein kinase B (p-AKT)] showed that the expression of p-PI3K(1.05±0.05) and p-AKT(1.17±0.06) in the IUA group increased compared to the control group (0.78±0.03, 0.85±0.05) (P=0.002, 0.002). The expression of p-PI3K (0.74±0.02) and p-AKT (0.93±0.04) proteins in the hUCB-MSC transplantation group decreased compared to the IUA group (P=0.003, 0.005). Conclusion: hUCB-MSC can repair the damaged endometrium in rabbit intrauterine adhesion model by increasing the number of endometrial glands and improving its level of fibrosis.

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来源期刊
Zhonghua yi xue za zhi
Zhonghua yi xue za zhi Medicine-Medicine (all)
CiteScore
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