RS4651 通过 TGF-β1/SMAD 信号通路抑制肺成纤维细胞的激活。

RS4651 suppresses lung fibroblast activation via the TGF-β1/SMAD signalling pathway.

机构信息

Department of Infectious Disease, Beijing Friendship Hospital, Capital Medical University, Beijing, 100050, China.

Experimental Center, Liver Research Center, Beijing Friendship Hospital, Capital Medical University, National Clinical Research Center for Digestive Disease, Beijing 100050, PR China.

出版信息

Eur J Pharmacol. 2021 Jul 15;903:174135. doi: 10.1016/j.ejphar.2021.174135. Epub 2021 May 1.

DOI:10.1016/j.ejphar.2021.174135

PMID:33940030

Abstract

BACKGROUND

Idiopathic pulmonary fibrosis (IPF) is a progressive disease resulting in respiratory failure with no efficient treatment options. We investigated the protective effect of RS4651 on pulmonary fibrosis in mice and the mechanism.

METHODS

Intratracheal injection of bleomycin (BLM) was used to induce pulmonary fibrosis in mice. RS4561 was administered intraperitoneally at different doses. Histopathological changes were observed. The level of alpha-smooth muscle actin (α-SMA) were also tested. In vitro, the proliferation and migratory effects of RS4651 treatment on MRC-5 cells pre-treated with transforming growth factor (TGF-β1) were examined. RNA-sequencing was used to detect differentially expressed target genes. Then, the expression of α-SMA, pSMAD2 and SMAD7 were analysed during RS4651 treatment of MRC-5 cells with or without silencing by SMAD7 siRNA.

RESULTS

Histopathological staining results showed decreased collagen deposition in RS4651 administered mice. Additionally, a lower level of α-SMA was also observed compared to the BLM group. The results of in vitro studies confirmed that RS4651 can inhibit the proliferation and migration, as well as α-SMA and pSMAD2 expression in MRC-5 cells treated with TGF-β1. RNA-sequencing data identified the target gene SMAD7. We found that RS4651 could upregulate SMAD7 expression and inhibit the proliferation and migration of MRC-5 cells via SMAD7, and RS4651 inhibition of α-SMA and pSMAD2 expression was blocked in SMAD7-siRNA MRC-5 cells. In vivo studies further confirmed that RS4651 could upregulate SMAD7 expression in BLM-induced lung fibrosis in mice.

CONCLUSIONS

Our data suggest that RS4651 alleviates BLM-induced pulmonary fibrosis in mice by inhibiting the TGF-β1/SMAD signalling pathway.

摘要

背景

特发性肺纤维化（IPF）是一种导致呼吸衰竭的进行性疾病，目前尚无有效的治疗方法。我们研究了 RS4651 对小鼠肺纤维化的保护作用及其机制。

方法

通过气管内注射博莱霉素（BLM）诱导小鼠发生肺纤维化，腹腔内给予不同剂量的 RS4651。观察组织病理学变化，检测α-平滑肌肌动蛋白（α-SMA）的水平。体外，观察 RS4651 对 TGF-β1 预处理的 MRC-5 细胞增殖和迁移的影响，进行 RNA 测序检测差异表达的靶基因，然后分析 RS4651 处理对 MRC-5 细胞中 SMAD7 沉默或不沉默时α-SMA、pSMAD2 和 SMAD7 的表达。

结果

组织学染色结果显示，RS4651 给药组胶原沉积减少。与 BLM 组相比，α-SMA 水平也较低。体外研究结果证实，RS4651 可抑制 TGF-β1 处理的 MRC-5 细胞的增殖和迁移，以及α-SMA 和 pSMAD2 的表达。RNA 测序数据鉴定出靶基因 SMAD7。我们发现 RS4651 可通过 SMAD7 上调 SMAD7 表达，抑制 MRC-5 细胞的增殖和迁移，并且在 SMAD7-siRNA MRC-5 细胞中 RS4651 抑制α-SMA 和 pSMAD2 的表达被阻断。体内研究进一步证实，RS4651 可上调 BLM 诱导的肺纤维化小鼠肺组织中 SMAD7 的表达。