Yu Hong Wei, Wang Wei Wei, Jing Qian, Pan Yong Liang
School of Medicine, Huzhou University, Huzhou, Zhejiang, China.
Am J Rhinol Allergy. 2023 Nov;37(6):739-750. doi: 10.1177/19458924231193154. Epub 2023 Aug 3.
Airway remodeling is demonstrated in Asian patients with allergic rhinitis (AR). The epithelial-mesenchymal transition (EMT) is one of the key mechanisms underlying airway remodeling. Thymic stromal lymphopoietin (TSLP) is an important contributor to airway remodeling. Although increased TSLP is found in AR, little is known about whether TSLP is involved in airway remodeling through induction of the EMT.
We investigated the effect of TSLP on the EMT in human nasal epithelial cells (HNECs) from AR patients.
Human nasal epithelial cells from AR patients were stimulated with TSLP in the absence or presence of the preincubation with a selective inhibitor of transforming growth factor beta 1 (TGF-β1) receptor (SB431542). The expression of TGF-β1 in the cells was evaluated by using real-time polymerase chain reaction, Western blotting, and immunocytochemistry. Western blotting and immunocytochemistry were used to assay EMT markers including vimentin, fibroblast-specific protein 1 (FSP1) and E-cadherin, small mothers against decapentaplegic homolog2/3 (Smad2/3), and phosphorylated Smad2/3 in the cells. The levels of extracellular matrix components such as collagens I and III in supernatants were measured by enzyme-linked immunoassay. Morphological changes of the cells were observed under inverted phase-contrast microscope.
A concentration-dependent increase of TGF-β1 mRNA and protein was observed following stimulation with TSLP. Furthermore, TSLP decreased the expression of E-cadherin protein, but upregulated the production of FSP1 and vimentin proteins along with increased levels of collagens I and III, and the morphology of the cells was transformed into fibroblast-like shape. Additionally, a significant increase was found in phosphorylation of Smad2/3 protein. However, these effects were reversed by SB431542 preincubation.
TSLP-induced HNECs to undergo the EMT process via TGF-β1-mediated Smad2/3 activation. TSLP is an activator of the EMT in HNECs and might be a potential target for inhibiting EMT and reducing airway remodeling in AR.
在亚洲过敏性鼻炎（AR）患者中已证实存在气道重塑。上皮-间质转化（EMT）是气道重塑的关键机制之一。胸腺基质淋巴细胞生成素（TSLP）是气道重塑的重要促成因素。虽然在AR患者中发现TSLP水平升高，但关于TSLP是否通过诱导EMT参与气道重塑却知之甚少。
我们研究了TSLP对AR患者人鼻上皮细胞（HNECs）中EMT的影响。
在存在或不存在转化生长因子β1（TGF-β1）受体选择性抑制剂（SB431542）预孵育的情况下，用TSLP刺激AR患者的人鼻上皮细胞。通过实时聚合酶链反应、蛋白质印迹法和免疫细胞化学法评估细胞中TGF-β1的表达。蛋白质印迹法和免疫细胞化学法用于检测细胞中的EMT标志物，包括波形蛋白、成纤维细胞特异性蛋白1（FSP1）和E-钙黏蛋白、小母亲抗五体不全同源蛋白2/3（Smad2/3）以及磷酸化Smad2/3。通过酶联免疫吸附测定法测量上清液中细胞外基质成分如I型和III型胶原蛋白的水平。在倒置相差显微镜下观察细胞的形态变化。
用TSLP刺激后，观察到TGF-β1 mRNA和蛋白质呈浓度依赖性增加。此外，TSLP降低了E-钙黏蛋白的表达，但上调了FSP1和波形蛋白的产生，同时I型和III型胶原蛋白水平增加，细胞形态转变为成纤维细胞样形状。此外，还发现Smad2/3蛋白的磷酸化显著增加。然而，SB431542预孵育可逆转这些作用。
TSLP通过TGF-β1介导的Smad2/3激活诱导HNECs发生EMT过程。TSLP是HNECs中EMT的激活剂，可能是抑制AR中EMT和减少气道重塑的潜在靶点。
