间充质基质细胞逆转子宫内膜纤维化的机制研究

Mechanism of mesenchymal stromal cells reversing fibrosis of the uterine endometrium after injury

摘要

为探讨间充质基质细胞（mesenchymal stromal cells, MSCs）对子宫内膜损伤小鼠模型纤维化进程及内膜再生的影响，采用物理搔刮联合脂多糖（LPS）化学诱导建立小鼠子宫内膜损伤模型，单侧给予 MSCs 后通过苏木精-伊红（HE）染色和 Masson 染色评估内膜形态与胶原沉积；采用 RT-qPCR 和免疫荧光法检测上皮标志物 Epcam 及胶原蛋白Ⅰ（Col1）的表达；提取原代子宫内膜上皮细胞，检测细胞周期蛋白 p21 的表达；通过转录组测序分析及 RT-qPCR 验证纤维化相关基因及信号通路。实验结果表明，MSCs 增加了子宫内膜厚度与腺体数量（P<0.05），并上调了Epcam 的表达（P<0.01），同时显著降低了子宫胶原纤维沉积（P<0.05）及原代上皮细胞中 p21 的表达（P<0.000 1）。转录组学分析表明，MSCs 通过调控 TGF-β 通路相关基因，抑制胶原沉积。本研究揭示了 MSCs 通过逆转子宫内膜纤维化、促进上皮再生，从而修复子宫损伤，其作用与调控 TGF-β/SMAD 通路、抑制胶原异常沉积有关。

Abstract

To investigate the effect of mesenchymal stromal cells (MSCs) on the fibrosis progression and endometrial regeneration in mouse models of endometrial injury, a mouse endometrial injury model was established by physical scraping combined with lipopolysaccharide (LPS) chemical induction, and the endometrium morphology and collagen deposition were evaluated by HE and Masson staining after unilateral administration of MSCs. RT-qPCR and immunofluorescence were used to detect the expression of epithelial markers Epcam and collagen I (Col1). Primary endometrial epithelial cells were isolated to assess p21 expression. Fibrosis-related genes and signaling pathways were validated using transcriptome sequencing and RT-qPCR. The experimental results showed that MSCs increased endometrial thickness and number of glands (P<0.05), up-regulated the expression ofEpcam (P<0.01), while significantly reducing uterine collagen fiber deposition (P<0.05) and the expression of p21 in primary epithelial cells (P<0.000 1). Transcriptomic analysis revealed that MSCs attenuated collagen deposition by modulating genes involved in the TGF-β signaling pathway. This study elucidates that MSCs facilitate uterine repair by reversing endometrial fibrosis and promoting epithelial regeneration, and their role is related to the regulation of the TGF-β/SMAD pathway and suppression of aberrant collagen accumulation.

关键词

子宫内膜损伤 / 间充质基质细胞 / 胶原纤维 / TGF-β/SMAD 通路

Key words

endometrial injury / mesenchymal stromal cells / collagen fiber / TGF-β/SMAD pathway

参考文献

[1] Li YF, Yin WM, Dong HQ, et al. Novel injectable sodium alginate hydrogel developed for improved endometrial repair with human umbilical cord mesenchymal stem cells[J]. Drug Deliv Transl Res, 2025, 15(12): 4638-4653.

[2] Hooker AB, Lemmers M, Thurkow AL, et al. Systematic review and meta—analysis of intrauterine adhesions after miscarriage: prevalence, risk factors and long—term reproductive outcome[J]. Hum Reprod Update, 2014, 20(2): 262-278.

[3] Ebrahim N, Mostafa O, El Dosoky RE, et al. Human mesenchymal stem cell—derived extracellular vesicles/estrogen combined therapy safely ameliorates experimentally induced intrauterine adhesions in a female rat model[J]. Stem Cell Res Ther, 2018, 9(1): 175.

[4] Wang J, Chen Q, Wang YY, et al. Multiresponsive microcapsules for prevention of intrauterine adhesion[J]. ACS Nano, 2025, 19(6): 6499-6510.

[5] Lin JY, Wang Z, Huang JL, et al. Microenvironment—protected exosome—hydrogel for facilitating endometrial regeneration, fertility restoration, and live birth of offspring[J]. Small, 2021, 17(11): 2007235.

[6] Yin Z, Wang J, Cui WG, et al. Advanced biomaterials for promoting endometrial regeneration[J]. Adv Healthc Mater, 2023, 12(16): e2202490.

[7] Kitaya K, Takeuchi T, Mizuta S, et al. Endometritis: new time, new concepts[J]. Fertil Steril, 2018, 110(3): 344-350.

[8] Shi YF, Wang Y, Li Q, et al. Immunoregulatory mechanisms of mesenchymal stem and stromal cells in inflammatory diseases[J]. Nat Rev Nephrol, 2018, 14(8): 493-507.

[9] Rodríguez—Eguren A, Bueno—Fernandez C, Gómez—Álvarez M, et al. Evolution of biotechnological advances and regenerative therapies for endometrial disorders: a systematic review[J]. Hum Reprod Update, 2024, 30(5): 584-613.

[10] Lu S, Xue HQ, Zhang FB, et al. Blastocyst structure—inspired core—shell stem cell microspheres with hyperoside delivery for endometrial regeneration[J]. Adv Healthc Mater, 2026, 15(6): e02293.

[11] Jiang XX, Li XM, Fei XW, et al. Endometrial membrane organoids from human embryonic stem cell combined with the 3D Matrigel for endometrium regeneration in asherman syndrome[J]. Bioact Mater, 2021, 6(11): 3935-3946.

[12] Song YT, Liu PC, Tan J, et al. Stem cell—based therapy for ameliorating intrauterine adhesion and endometrium injury[J]. Stem Cell Res Ther, 2021, 12(1): 556.

[13] Song MM, Zhao GF, Sun HX, et al. circPTPN12/miR—21—5 p/∆Np63α pathway contributes to human endometrial fibrosis[J]. eLife, 2021, 10: e65735.

[14] Park M, Hong SH, Park SH, et al. Perivascular stem cell—derived cyclophilin a improves uterine environment with asherman's syndrome via HIF1α—dependent angiogenesis[J]. Mol Ther, 2020, 28(8): 1818-1832.

[15] Zhang YL, Lin XN, Dai YD, et al. Endometrial stem cells repair injured endometrium and induce angiogenesis via AKT and ERK pathways[J]. Reproduction, 2016, 152(5): 389-402.

[16] Chen MY, Liu YH, Cao Y, et al. Remodeling the proinflammatory microenvironment in osteoarthritis through interleukin—1 beta tailored exosome cargo for inflammatory regulation and cartilage regeneration[J]. ACS Nano, 2025, 19(4): 4924-4941.

[17] Yuan CS, Zhang Y, He YH, et al. Pantothenic acid alleviates endometrial injury induced by vagotomy in mice[J]. Mucosal Immunol, 2026, 19(1): 1573-1586.

[18] Mengru Wei . Pharmacodynamics and Pharmacokinetics of mesenchymal stem cells in damaged uterus[D]. Nanjing: China Pharmaceutical University, 2023.

[19] Ang CJ, Skokan TD, McKinley KL. Mechanisms of regeneration and fibrosis in the endometrium[J]. Annu Rev Cell Dev Biol, 2023, 39: 197-221.

[20] Munro MG, Salazar CA, Bhagavath B, et al. The epidemiology, clinical burden, and prevention of intrauterine adhesions (IUAs) related to surgically induced endometrial trauma: a systematic literature review and selective meta—analyses[J]. Hum Reprod Update, 2025, 31(6): 588-625.

[21] Shi Q, Gao JW, Jiang Y, et al. Differentiation of human umbilical cord Wharton's jelly—derived mesenchymal stem cells into endometrial cells[J]. Stem Cell Res Ther, 2017, 8(1): 246.

[22] Marquardt RM, Grimm SA, Wu SP, et al. Serum response factor is essential for endometrial function and prevention of inflammatory fibrosis[J]. Proc Natl Acad Sci USA, 2025, 122(44): e2510060122.

[23] Zhou YJ, Geng YQ, Gao RF, et al. Early pregnancy exposure to beta—cypermethrin compromises endometrial decidualisation in mice via downregulation of cyclin D3, CDK4/6, and p21[J]. Food Chem Toxicol, 2022, 169: 113382.

[24] Sun Q, Zhang D, Ai QY, et al. Human umbilical cord mesenchymal stem cells improve uterine incision healing after cesarean delivery in rats by modulating the TGF—β/Smad signaling pathway[J]. Arch Gynecol Obstet, 2024, 310(1): 103-111.

[25] Yang HW, Zhang WM, Fang JC, et al. Intrauterine infusion of platelet—rich plasma improves fibrosis by transforming growth factor beta 1/Smad pathway in a rat intrauterine adhesion model[J]. Reprod Biol, 2024, 24(2): 100882.

[26] Hou CY, Zhu HL, Chang XH. Mesenchymal stem cells: opening a new chapter in the treatment of gynecological diseases[J]. Stem Cell Res Ther, 2025, 16(1): 520.

[27] Ushakov RE, Burova EB. Conditioned medium of human mesenchymal stromal/stem cells cultured on decellularized extracellular matrix promotes murine skeletal muscle repair after acute injury[J]. Biochem Biophys Res Commun, 2024, 736: 150511.

基金资助

国家自然科学基金项目(82173882)

中国科协青年人才托举项目(2023QNRC001)

江苏省自然科学基金项目(BK20220149)