Matsuno Kei, Harada Norihiro, Harada Sonoko, Takeshige Tomohito, Ishimori Ayako, Itoigawa Yukinari, Katsura Yoko, Kodama Yuzo, Makino Fumihiko, Ito Jun, Atsuta Ryo, Akiba Hisaya, Takahashi Kazuhisa
a Department of Respiratory Medicine, Faculty of Medicine , Graduate School of Medicine, Juntendo University , Tokyo , Japan.
b Research Institute for Diseases of Old Ages, Faculty of Medicine , Graduate School of Medicine, Juntendo University , Tokyo , Japan.
Exp Lung Res. 2018 Sep;44(7):332-343. doi: 10.1080/01902148.2018.1522558. Epub 2019 Jan 24.
In patients with asthma, chronic inflammatory processes and the subsequent remodeling of the airways contribute to the symptoms and the pathophysiological changes. Epithelial-mesenchymal transition (EMT) is thought to play an important role in tissue remodeling. Previous reports show that tumor necrosis factor (TNF)-like weak inducer of apoptosis (TWEAK) is a cytokine of the TNF superfamily, exerts pro-inflammatory effects, and enhances transforming growth factor (TGF)-β-induced EMT in bronchial epithelial cells. In this study, we investigated the TWEAK-induced cytokine and chemokine production in the human bronchial epithelial cell line BEAS-2B during EMT.
Quantitative real-time RT-PCR, enzyme-linked immunosorbent assays, western blotting, and immunohistochemistry were used to define the production of cytokines and chemokines.
We found that TWEAK increases mRNA and protein levels of thymic stromal lymphopoietin (TSLP), monocyte chemoattractant protein -1 (MCP-1), regulated upon activation normal T cell express sequence (RANTES), and IL-8 in BEAS-2B bronchial epithelial cells. Moreover, co-treatment with TWEAK and TGF-β1 induces not only features of EMT but also enhances the production of TSLP and RANTES. Thymus- and activation-regulated chemokines (TARC) production is induced by the co-treatment of TWEAK and TGF-β1 but not by TWEAK or TGF-β1 stimulation alone. Furthermore, the increased mRNA expression of TSLP and RANTES after co-treatment with TWEAK and TGF-β1 is prevented by inhibitors of Smad-independent signaling pathways.
In the present study, we have revealed a novel mechanism for the production of asthma-related cytokines and chemokines in EMT driven by the co-stimulation with TWEAK and TGF-β1. We conclude that cellular EMT processes caused by TWEAK and TGF-β1 may contribute to chronic airway inflammation and remodeling.
在哮喘患者中，慢性炎症过程及随后的气道重塑导致了症状和病理生理变化。上皮-间质转化（EMT）被认为在组织重塑中起重要作用。既往报道显示，肿瘤坏死因子（TNF）-凋亡微弱诱导因子（TWEAK）是TNF超家族的一种细胞因子，具有促炎作用，并增强支气管上皮细胞中转化生长因子（TGF）-β诱导的EMT。在本研究中，我们调查了TWEAK诱导人支气管上皮细胞系BEAS-2B在EMT过程中细胞因子和趋化因子的产生情况。
采用定量实时RT-PCR、酶联免疫吸附测定、蛋白质印迹法和免疫组织化学法来确定细胞因子和趋化因子的产生。
我们发现TWEAK可增加BEAS-2B支气管上皮细胞中胸腺基质淋巴细胞生成素（TSLP）、单核细胞趋化蛋白-1（MCP-1）、活化正常T细胞表达和分泌因子（RANTES）以及IL-8的mRNA和蛋白质水平。此外，TWEAK与TGF-β1联合处理不仅诱导了EMT特征，还增强了TSLP和RANTES的产生。TWEAK与TGF-β1联合处理可诱导胸腺和活化调节趋化因子（TARC）的产生，但单独的TWEAK或TGF-β1刺激则不能。此外，TWEAK与TGF-β1联合处理后TSLP和RANTES mRNA表达的增加可被Smad非依赖信号通路抑制剂所阻断。
在本研究中，我们揭示了一种由TWEAK和TGF-β1共同刺激驱动的EMT过程中哮喘相关细胞因子和趋化因子产生的新机制。我们得出结论，TWEAK和TGF-β1引起的细胞EMT过程可能导致慢性气道炎症和重塑。
