Zhang Haiwei, Yang Hui, Wu Yingyi, Shi Yirui, Xu Min, Zhang Yueyang, Chen Hongwei, Sun Lingyun
Department of Rheumatology and Immunology, Nanjing Drum Tower Hospital, Clinical College of Xuzhou Medical University, Nanjing, China.
Department of Rheumatology and Immunology, Nanjing Drum Tower Hospital, Clinical College of Nanjing University of Chinese Medicine, Nanjing, China.
Lupus Sci Med. 2025 Aug 5;12(2):e001468. doi: 10.1136/lupus-2024-001468.
SLE is a multisystem autoimmune disease characterised by chronic inflammation and progressive organ damage, including ovarian dysfunction. This study investigated the therapeutic efficacy of umbilical cord-derived mesenchymal stem cells (UC-MSCs) in ameliorating ovarian impairment and restoring ovarian function through the inhibition of fibrosis in a lupus mouse model.
Serum levels of sex hormones were quantified via ELISA. Ovarian tissue samples were histologically evaluated for follicle count and fibrosis via H&E and Masson's trichrome staining. Quantitative reverse-transcriptase-PCR, western blot, immunofluorescence and immunohistochemistry were employed to evaluate inflammatory cytokines, fibrotic factors, hormone receptors and signalling proteins. Primary granulosa cells (GCs) isolated from lupus mice (MRL/lpr) were cocultured with MSCs and the expression of fibrotic factors was analysed by western blot. Additionally, a human GC line (KGN) was used to further explore the relationships among connective tissue growth factor (CTGF), focal adhesion kinase (FAK)/FAK-Tyr576/577 phosphorylation and fibrosis. This was achieved through stimulation with recombinant CTGF, the CTGF antagonist FG-3019 or the FAK inhibitor SU6656.
UC-MSC transplantation significantly downregulated the expression of proinflammatory cytokines (, ) and fibrotic markers (, ) while upregulating the expression of key hormone receptors (, , ). Additionally, a reduction in CD3/CD4 T-cell infiltration, C3 complement deposition and IgG levels was observed, accompanied by an increase in regulatory T cells. Further analysis revealed that fibrotic markers and FAK-Tyr576/577 phosphorylation were markedly suppressed in primary ovarian GCs following MSC transplantation. In vitro experiments demonstrated that recombinant CTGF promoted fibrogenesis in the human GC line KGN. Conversely, MSC treatment inhibited phosphorylated FAK-Tyr576/577 and downregulated the expression of Collagen 1 and α-SMA, suggesting that UC-MSCs alleviate ovarian fibrosis by suppressing FAK-Tyr576/577 phosphorylation.
This study demonstrated that UC-MSC treatment ameliorated ovarian dysfunction and attenuated ovarian fibrosis in lupus mice by modulating the CTGF/FAK-Tyr576/577 phosphorylation pathway.
系统性红斑狼疮（SLE）是一种多系统自身免疫性疾病，其特征为慢性炎症和进行性器官损害，包括卵巢功能障碍。本研究通过抑制狼疮小鼠模型中的纤维化，探讨脐带间充质干细胞（UC-MSCs）在改善卵巢损伤和恢复卵巢功能方面的治疗效果。
通过酶联免疫吸附测定（ELISA）对血清性激素水平进行定量。通过苏木精-伊红（H&E）染色和马松三色染色对卵巢组织样本进行组织学评估，以计数卵泡和检测纤维化情况。采用定量逆转录聚合酶链反应（qRT-PCR）、蛋白质印迹法、免疫荧光法和免疫组织化学法评估炎性细胞因子、纤维化因子、激素受体和信号蛋白。将从狼疮小鼠（MRL/lpr）分离的原代颗粒细胞（GCs）与间充质干细胞共培养，并通过蛋白质印迹法分析纤维化因子的表达。此外，使用人GC系（KGN）进一步探讨结缔组织生长因子（CTGF）、粘着斑激酶（FAK）/FAK-Tyr576/577磷酸化与纤维化之间的关系。这是通过用重组CTGF、CTGF拮抗剂FG-3019或FAK抑制剂SU6656刺激来实现的。
UC-MSC移植显著下调促炎细胞因子（……）和纤维化标志物（……）的表达，同时上调关键激素受体（……）的表达。此外，可以观察到CD3/CD4 T细胞浸润减少、C3补体沉积和IgG水平降低，同时调节性T细胞增加。进一步分析表明移植间充质干细胞后，原代卵巢颗粒细胞中的纤维化标志物和FAK-Tyr576/577磷酸化受到显著抑制。体外实验表明重组CTGF促进人GC系KGN中的纤维生成。相反，间充质干细胞处理抑制了磷酸化的FAK-Tyr576/577，并下调了胶原蛋白1和α-平滑肌肌动蛋白（α-SMA）的表达，表明UC-MSCs通过抑制FAK-Tyr576/577磷酸化减轻卵巢纤维化。
本研究表明，UC-MSC治疗通过调节CTGF/FAK-Tyr576/577磷酸化途径改善了狼疮小鼠的卵巢功能障碍并减轻了卵巢纤维化。
