Abstract
Cardiac fibrosis is a cause of morbidity and mortality in people with heart disease. Anti-fibrosis treatment is a significant therapy for heart disease, but there is still no thorough understanding of fibrotic mechanisms. This study was carried out to ascertain the functions of cytokine receptor-like factor 1 (CRLF1) in cardiac fibrosis and clarify its regulatory mechanisms. We found that CRLF1 was expressed predominantly in cardiac fibroblasts. Its expression was up-regulated not only in a mouse heart fibrotic model induced by myocardial infarction, but also in mouse and human cardiac fibroblasts provoked by transforming growth factor-β1 (TGF-β1). Gain- and loss-of-function experiments of CRLF1 were carried out in neonatal mice cardiac fibroblasts (NMCFs) with or without TGF-β1 stimulation. CRLF1 overexpression increased cell viability, collagen production, cell proliferation capacity, and myofibroblast transformation of NMCFs with or without TGF-β1 stimulation, while silencing of CRLF1 had the opposite effects. An inhibitor of the extracellular signal-regulated kinase 1/2 (ERK1/2) signaling pathway and different inhibitors of TGF-β1 signaling cascades, comprising mothers against decapentaplegic homolog (SMAD)-dependent and SMAD-independent pathways, were applied to investigate the mechanisms involved. CRLF1 exerted its functions by activating the ERK1/2 signaling pathway. Furthermore, the SMAD-dependent pathway, not the SMAD-independent pathway, was responsible for CRLF1 up-regulation in NMCFs treated with TGF-β1. In summary, activation of the TGF-β1/SMAD signaling pathway in cardiac fibrosis increased CRLF1 expression. CRLF1 then aggravated cardiac fibrosis by activating the ERK1/2 signaling pathway. CRLF1 could become a novel potential target for intervention and remedy of cardiac fibrosis.
摘要
心脏纤维化是心脏疾病患者发病和死亡的原因之一. 抗纤维化治疗是一种治疗心脏疾病的重要手段, 但目前对纤维化的机制仍缺乏深入了解. 本研究旨在确定细胞因子受体样因子1(CRLF1)在心脏纤维化中的功能并阐明其调节机制. 我们发现CRLF1主要在心脏成纤维细胞中表达; 无论是在心肌梗死诱导的小鼠心脏纤维化模型还是在转化生长因子-β1(TGF-β1)刺激的小鼠和人心脏成纤维细胞中, CRLF1表达均上调. 本研究在使用或不使用TGF-β1刺激的新生乳鼠心脏成纤维细胞(NMCFs)中开展了CRLF1的功能获得和丧失实验. 在TGF-β1刺激或不刺激的情况下, CRLF1的过表达均可增加NMCFs的细胞活力、 胶原生成、 细胞增殖能力及肌成纤维细胞转化, 而CRLF1沉默则具有相反效果. 应用细胞外信号调节激酶1/2(ERK1/2)信号通路抑制剂以及包括SMAD依赖和非依赖信号在内的不同TGF-β1下游信号通路抑制剂来开展机制研究. CRLF1通过激活ERK1/2信号通路发挥其功能. 此外, CRLF1在TGF-β1处理的NMCFs中表达上调是由SMAD依赖性通路介导, 而不是SMAD非依赖性通路介导. 总而言之, 心脏纤维化中TGF-β1/SMAD信号通路的激活增加了CRLF1的表达. 随后, CRLF1通过激活ERK1/2信号通路加重了心脏纤维化. 因此, CRLF1可作为一个干预和治疗心脏纤维化的新的潜在靶点.
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Acknowledgments
This work was supported by the National Key Research and Development Project of China (No. 2018YFA0800404), the National Natural Science Foundation of China (Nos. 82100255 and 81970736), and the China Postdoctoral Science Foundation (Nos. 2021M691459 and 2022T150299).
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Huijie ZHANG, Shenjian LUO, and Jin LI contributed to conception and designed this study. Zhi YANG and Ruxin CHEN carried out the experiments. Shenjian LUO performed the statistical analysis and revised the manuscript. Danming YOU, Fei TENG, and Wenhui LIU wrote the first manuscript version. Shenjian LUO, Zhi YANG, and Ruxin CHEN discussed and modified all sections of the manuscript. Youwen YUAN supervised the experiments. All authors have reviewed and approved the final manuscript, and therefore, have full access to all the data in the study and take responsibility for the integrity and security of the data.
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Shenjian LUO, Zhi YANG, Ruxin CHEN, Danming YOU, Fei TENG, Youwen YUAN, Wenhui LIU, Jin LI, and Huijie ZHANG declare that they have no conflict of interest.
The animal experiments were carried out under the approval of the Institutional Animal Care and Use Committee of Nanfang Hospital, Southern Medical University (No.NFYY-2021-1047).
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Luo, S., Yang, Z., Chen, R. et al. Cytokine receptor-like factor 1 (CRLF1) promotes cardiac fibrosis via ERK1/2 signaling pathway. J. Zhejiang Univ. Sci. B 24, 682–697 (2023). https://doi.org/10.1631/jzus.B2200506
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DOI: https://doi.org/10.1631/jzus.B2200506
Key words
- Cytokine receptor-like factor 1 (CRLF1)
- TGF-β1/SMAD signaling pathway
- ERK1/2 signaling pathway
- Cardiac fibrosis
- Myofibroblast transformation
- Extracellular matrix (ECM)