Department of Rheumatology and Immunology, The Second Xiangya Hospital, Central South University, Changsha, China.
Clinical Immunology Research Center, Central South University, Changsha, China.
Eur J Clin Invest. 2022 Aug;52(8):e13791. doi: 10.1111/eci.13791. Epub 2022 Apr 27.
Iguratimod (T-614), exerting a powerful anti-inflammatory ability, has therapeutic efficacy in multiple autoimmune diseases. However, the effect of T-614 on systemic sclerosis (SSc) is unclear. Here, we investigate the effect and molecular mechanism of T-614 in experimental SSc models.
In vitro, cultured dermal fibroblasts from four SSc patients were subjected to different doses of T-614 in the presence or absence of TGF-β1 stimulation. Cell proliferation, apoptosis and migration were determined by CCK-8, flow cytometry and transwell assay, respectively. Fibrosis markers and smad signalling pathway-related proteins were detected by immunoblotting and immunofluorescence. In vivo, a bleomycin-induced SSc mouse model was used to evaluate the effect of T-614 on skin fibrosis. Pathological changes in skin tissues were evaluated by HE, Masson staining and immunohistochemistry.
In the study, we found T-614 inhibited TGF-β1-induced cell proliferation, migration and promoted apoptosis in a dose-dependent manner (all p < 0.01). T-614 partially reversed TGF-β1-induced upregulation of fibrosis markers and phosphorylation of smad2 and smad3 and blocked p-Smad3 nuclear translocation (all p < 0.05), suggesting T-614 may inhibit dermal fibroblasts activation by regulating TGF-β1/smad pathway. In vivo experiments, T-614 alleviated skin thickness in bleomycin-induced SSc mice (all p < 0.05). The expression of fibrosis markers and the infiltration of macrophages in skin tissue were significantly decreased after T-614 treatment (all p < 0.05).
Our preliminary data indicated T-614 inhibited dermal fibroblasts activation and skin fibrosis at least partly by regulating TGF-β1/smad pathway in experimental SSc models and may be a promising therapeutic agent for SSc.
昔布特罗（T-614）具有强大的抗炎能力，在多种自身免疫性疾病中具有治疗效果。然而，T-614 对系统性硬化症（SSc）的作用尚不清楚。在这里，我们研究了 T-614 在实验性 SSc 模型中的作用和分子机制。
在体外，来自四名 SSc 患者的培养真皮成纤维细胞在存在或不存在 TGF-β1 刺激的情况下接受不同剂量的 T-614。通过 CCK-8、流式细胞术和 Transwell 测定分别测定细胞增殖、凋亡和迁移。通过免疫印迹和免疫荧光法检测纤维化标志物和 smad 信号通路相关蛋白。在体内，使用博来霉素诱导的 SSc 小鼠模型评估 T-614 对皮肤纤维化的影响。通过 HE、Masson 染色和免疫组化评估皮肤组织的病理变化。
在研究中，我们发现 T-614 以剂量依赖性方式抑制 TGF-β1 诱导的细胞增殖、迁移并促进凋亡（均 p<0.01）。T-614 部分逆转 TGF-β1 诱导的纤维化标志物上调和 smad2 和 smad3 的磷酸化，并阻止 p-Smad3 核转位（均 p<0.05），表明 T-614 可能通过调节 TGF-β1/smad 通路抑制真皮成纤维细胞的激活。体内实验中，T-614 减轻博来霉素诱导的 SSc 小鼠的皮肤厚度（均 p<0.05）。T-614 治疗后皮肤组织中纤维化标志物的表达和巨噬细胞的浸润明显减少（均 p<0.05）。
我们的初步数据表明，T-614 通过调节实验性 SSc 模型中的 TGF-β1/smad 通路至少部分抑制真皮成纤维细胞的激活和皮肤纤维化，可能是 SSc 的一种有前途的治疗药物。
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