The novel inhibitor PRI-724 for Wnt/β-catenin/CBP signaling ameliorates bleomycin-induced pulmonary fibrosis in mice
The novel inhibitor PRI-724 for Wnt/β-catenin/CBP signaling ameliorates bleomycin-induced pulmonary fibrosis in mice
复制标题
DOI：
10.1080/01902148.2019.1638466
复制
发表时间：
2019-07-12
影响因子：
1.7
通讯作者：
Nishioka, Yasuhiko
中科院分区：
文献类型：
--
作者：
Okazaki, Hiroyasu;Sato, Seidai;Nishioka, Yasuhiko
研究目的/目的：据报道，Wnt/β-连环蛋白信号传导在肺纤维化中被激活，并且被集中作为抗纤维化治疗的靶点。然而，抑制Wnt/β-catenin信号通路如何改善肺纤维化的机制尚未完全阐明。本研究的目的是探索Wnt/beta-catenin抑制剂在肺纤维化中的靶细胞，并检测特异性阻断beta-catenin和CBP相互作用的新型抑制剂PRI-724的抗纤维化作用。材料与方法：在成纤维细胞和巨噬细胞上检查了PRI-724的活性代谢物C-82对TGF-β 1和α-平滑肌肌动蛋白（SMA）表达的影响。我们还检查了PRI-724在博来霉素诱导的肺纤维化小鼠模型中的作用。结果如下：使用蛋白质印迹法在肺成纤维细胞以及由Wnt 3a刺激的巨噬细胞中检测到经典途径中β-连环蛋白的活化和增加的积累。用C-82处理减少了CBP蛋白，增加了肺成纤维细胞核中p300蛋白与β-连环蛋白的结合。此外，C-82抑制用TGF-β处理的肺成纤维细胞中SMA的表达，表明肌成纤维细胞分化的抑制。在博莱霉素诱导的纤维化肺中，β-连环蛋白在巨噬细胞中染色强烈，而在肺泡上皮细胞和成纤维细胞中染色较弱。当以晚期而非早期治疗方案施用时，PRI-724的施用改善了小鼠中由博来霉素诱导的肺纤维化。支气管肺泡液（BALF）分析显示肺泡巨噬细胞数量减少。此外，在用PRI-724处理的小鼠中，BALF中TGF-β 1的水平降低。C-82还抑制肺泡巨噬细胞产生TGF-β 1。结论：这些结果表明，β-连环蛋白/CBP抑制剂PRI-724是一种有效的抗纤维化药物，通过调节肺中巨噬细胞的活性发挥作用。
Purpose/Aim of the Study: Wnt/beta-catenin signaling was reported to be activated in pulmonary fibrosis, and was focused on as a target for antifibrotic therapy. However, the mechanism how the inhibition of Wnt/beta-catenin signaling ameliorate pulmonary fibrosis has not been fully elucidated. The purpose of this study is to explore the target cells of Wnt/beta-catenin inhibition in pulmonary fibrosis and to examine the antifibrotic effect of the novel inhibitor PRI-724 specifically disrupting the interaction of beta-catenin and CBP. Materials and Methods: The effect of C-82, an active metabolite of PRI-724, on the expression of TGF-beta 1 and alpha-smooth muscle actin (SMA) was examined on fibroblasts and macrophages. We also examined the effects of PRI-724 in mouse model of bleomycin-induced pulmonary fibrosis. Results: The activation and increased accumulation of beta-catenin in the canonical pathway were detected in lung fibroblasts as well as macrophages stimulated by Wnt3a using Western blotting. Treatment with C-82 reduced CBP protein and increased p300 protein binding to beta-catenin in the nucleus of lung fibroblasts. In addition, C-82 inhibited the expression of SMA in lung fibroblasts treated with TGF-beta, indicating the inhibition of myofibroblast differentiation. In the fibrotic lungs induced by bleomycin, beta-catenin was stained strongly in macrophages, but the staining of beta-catenin in alveolar epithelial cells and fibroblasts was weak. The administration of PRI-724 ameliorated pulmonary fibrosis induced by bleomycin in mice when administered with a late, but not an early, treatment schedule. Analysis of bronchoalveolar fluid (BALF) showed a decreased number of alveolar macrophages. In addition, the level of TGF-beta 1 in BALF was decreased in mice treated with PRI-724. C-82 also inhibited the production of TGF-beta 1 by alveolar macrophages. Conclusions: These results suggest that the beta-catenin/CBP inhibitor PRI-724 is a potent antifibrotic agent that acts by modulating the activity of macrophages in the lungs.