尼达尼布可下调培养的系统性硬化症成纤维细胞向肌成纤维细胞的转化及其促纤维化活性。
Nintedanib downregulates the transition of cultured systemic sclerosis fibrocytes into myofibroblasts and their pro-fibrotic activity.
机构信息
Laboratory of Experimental Rheumatology and Academic Division of Clinical Rheumatology, Department of Internal Medicine, University of Genova, IRCCS San Martino Polyclinic Hospital, Genoa, Italy.
Department of Rheumatology, Ghent University Hospital, Ghent, Belgium.
出版信息
Arthritis Res Ther. 2021 Aug 3;23(1):205. doi: 10.1186/s13075-021-02555-2.
BACKGROUND
Circulating fibrocytes are an important source of fibroblasts and myofibroblasts, which are involved in fibrotic processes, including systemic sclerosis (SSc). The study aimed to investigate the effect of nintedanib (a tyrosine kinase inhibitor) in inhibiting the in vitro transition of circulating SSc fibrocytes into myofibroblasts and their pro-fibrotic activity.
METHODS
Circulating fibrocytes were obtained from 18 SSc patients and 5 healthy subjects (HSs). Cultured SSc fibrocytes were maintained in growth medium (untreated cells) or treated with nintedanib 0.1 and 1 μM for 3 and 24 h. Fibroblast-specific protein-1 (S100A4) and α-smooth muscle actin (αSMA), as markers of fibroblast/myofibroblast phenotype, together with type I collagen (COL1) and fibronectin (FN), were investigated by qRT-PCR and Western blotting. Non-parametric tests were used for statistical analysis.
RESULTS
Significantly elevated gene and protein expressions of αSMA, S100A4, COL1, and FN were observed in SSc fibrocytes compared to HS fibrocytes (gene: αSMA p < 0.001; others p < 0.0001; protein: all p < 0.05). Interestingly, an increased gene and protein expression of αSMA and S100A4 was found in fibrocytes from SSc patients positive for anti-Scl70 and with interstitial lung disease (ILD) (Scl70ILD) compared to Scl70ILD patients (S100A4: gene: p < 0.01; protein: p < 0.05), whereas no differences were observed for COL1 and FN. Nintedanib reduced gene and protein expression of αSMA, S100A4, COL1, and FN in SSc fibrocytes compared to untreated ones with different statistical significance. Noteworthy, nintedanib significantly downregulated gene and protein expression of αSMA, S100A4, COL1, and FN in Scl70ILD fibrocytes (all p < 0.05), whereas only that of S100A4 and FN was significantly downregulated (p < 0.05) in Scl70ILD fibrocytes compared to the related untreated cells.
CONCLUSIONS
Nintedanib seems to downregulate in vitro the transition of fibrocytes into myofibroblasts and their pro-fibrotic activity, particularly in cells isolated from Scl70ILD SSc patients.
背景
循环成纤维细胞是成纤维细胞和肌成纤维细胞的重要来源，参与纤维化过程，包括系统性硬皮病（SSc）。本研究旨在探讨尼达尼布（一种酪氨酸激酶抑制剂）在抑制循环 SSc 成纤维细胞向肌成纤维细胞转化及其促纤维化活性中的作用。
方法
从 18 例 SSc 患者和 5 例健康对照（HS）中获得循环成纤维细胞。培养的 SSc 成纤维细胞在生长培养基中（未处理的细胞）或用尼达尼布 0.1 和 1μM 处理 3 和 24 小时。通过 qRT-PCR 和 Western blot 检测成纤维细胞/肌成纤维细胞表型的标志，如纤维母细胞特异性蛋白-1（S100A4）和α-平滑肌肌动蛋白（αSMA），以及 I 型胶原（COL1）和纤维连接蛋白（FN）。采用非参数检验进行统计学分析。
结果
与 HS 成纤维细胞相比，SSc 成纤维细胞中 αSMA、S100A4、COL1 和 FN 的基因和蛋白表达显著升高（基因：αSMA p < 0.001；其他 p < 0.0001；蛋白：均 p < 0.05）。有趣的是，与 SSc 患者中抗 Scl70 阳性且伴有间质性肺病（ILD）（Scl70ILD）的患者相比，Scl70ILD 患者的成纤维细胞中 αSMA 和 S100A4 的基因和蛋白表达增加（S100A4：基因：p < 0.01；蛋白：p < 0.05），而 COL1 和 FN 则无差异。与未处理的细胞相比，尼达尼布降低了 SSc 成纤维细胞中 αSMA、S100A4、COL1 和 FN 的基因和蛋白表达，差异具有统计学意义。值得注意的是，尼达尼布显著下调了 Scl70ILD 成纤维细胞中 αSMA、S100A4、COL1 和 FN 的基因和蛋白表达（均 p < 0.05），而仅下调了 Scl70ILD 成纤维细胞中 S100A4 和 FN 的基因和蛋白表达（均 p < 0.05）与相关的未处理细胞相比。
结论
尼达尼布似乎可下调体外成纤维细胞向肌成纤维细胞的转化及其促纤维化活性，尤其是在 Scl70ILD SSc 患者分离的细胞中。
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