间充质干细胞通过CTGF/FAK信号抑制系统性红斑狼疮纤维化,改善卵巢功能。

IF 3.5 2区 医学 Q1 RHEUMATOLOGY
Haiwei Zhang, Hui Yang, Yingyi Wu, Yirui Shi, Min Xu, Yueyang Zhang, Hongwei Chen, Lingyun Sun
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引用次数: 0

摘要

目的:SLE是一种多系统自身免疫性疾病,以慢性炎症和进行性器官损害为特征,包括卵巢功能障碍。本研究探讨了脐带间充质干细胞(UC-MSCs)在狼疮小鼠模型中通过抑制纤维化改善卵巢损伤和恢复卵巢功能的治疗效果。方法:采用ELISA法测定血清性激素水平。通过H&E和马松三色染色对卵巢组织样本进行卵泡计数和纤维化的组织学评估。采用定量逆转录酶链式pcr、免疫印迹、免疫荧光和免疫组织化学检测炎症因子、纤维化因子、激素受体和信号蛋白。将从狼疮小鼠分离的原代颗粒细胞(MRL/lpr)与MSCs共培养,采用western blot方法分析其纤维化因子的表达。此外,利用人GC细胞系(KGN)进一步探讨结缔组织生长因子(CTGF)、局灶黏附激酶(FAK)/FAK- tyr576 /577磷酸化与纤维化的关系。这是通过重组CTGF、CTGF拮抗剂FG-3019或FAK抑制剂SU6656刺激实现的。结果:UC-MSC移植显著下调促炎因子(Tnf-α、Il-1β)和纤维化标志物(Ctgf、α-Sma)的表达,上调关键激素受体(Amh、Esr1、Esr2)的表达。此外,观察到CD3+/CD4+ T细胞浸润,C3补体沉积和IgG水平降低,并伴有调节性T细胞的增加。进一步分析显示,MSC移植后,纤维化标志物和FAK-Tyr576/577磷酸化在原发卵巢GCs中明显抑制。体外实验表明,重组CTGF促进了人GC细胞系KGN的纤维生成。相反,MSC处理抑制磷酸化的FAK-Tyr576/577,下调胶原1和α-SMA的表达,表明UC-MSCs通过抑制FAK-Tyr576/577磷酸化来减轻卵巢纤维化。结论:本研究表明UC-MSC治疗可通过调节CTGF/FAK-Tyr576/577磷酸化通路改善狼疮小鼠卵巢功能障碍,减轻卵巢纤维化。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Mesenchymal stem cells improve ovarian function by suppressing fibrosis through CTGF/FAK signalling in systemic lupus erythematosus.

Objective: 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.

Methods: 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.

Results: UC-MSC transplantation significantly downregulated the expression of proinflammatory cytokines (Tnf-α, Il-1β) and fibrotic markers (Ctgf, α-Sma) while upregulating the expression of key hormone receptors (Amh, Esr1, Esr2). 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.

Conclusion: 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.

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来源期刊
Lupus Science & Medicine
Lupus Science & Medicine RHEUMATOLOGY-
CiteScore
5.30
自引率
7.70%
发文量
88
审稿时长
15 weeks
期刊介绍: Lupus Science & Medicine is a global, peer reviewed, open access online journal that provides a central point for publication of basic, clinical, translational, and epidemiological studies of all aspects of lupus and related diseases. It is the first lupus-specific open access journal in the world and was developed in response to the need for a barrier-free forum for publication of groundbreaking studies in lupus. The journal publishes research on lupus from fields including, but not limited to: rheumatology, dermatology, nephrology, immunology, pediatrics, cardiology, hepatology, pulmonology, obstetrics and gynecology, and psychiatry.
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