Department of Otolaryngology-Head and Neck Surgery, Kyoto Prefectural University of Medicine, Kyoto, Japan.
Department of Otolaryngology-Head and Neck Surgery, NYU Grossman School of Medicine, New York, New York, USA.
Laryngoscope. 2024 Nov;134(11):4593-4598. doi: 10.1002/lary.31570. Epub 2024 Jun 11.
Vocal fold scar remains a therapeutic challenge. Vocal fold fibroblasts (VFFs) secrete extracellular matrix (ECM), and transforming growth factor-beta 1 (TGF-β1)-mediated fibroblast to myofibroblast differentiation is central to the development of fibrosis. The transient receptor potential (TRP) channel superfamily is a group of nonselective cation channels, and activation of TRP ankyrin 1 (TRPA1) channel has been shown to have antifibrotic effects through TGF-β1/Smad signaling in various organs. This study aimed to elucidate expression of TRPA1 and the impact of TRPA1 activation on TGF-β1/Smad signaling in VFFs.
Vocal folds were dissected from 10-week-old, male Sprague-Dawley rats and primary VFFs were established. TRPA1 was examined in VFFs and lamina propria via immunostaining. VFFs were treated with allyl isothiocyanate (AITC, TRP channel agonist, 10 M) ± TGF-β1 (10 ng/ml) ± A-967079 (selective TRPA1 channel antagonist, 5.0 × 10 M) for 4 or 24 h. Trpa1, Smad3, Smad7, Col1a1, Acta2, and Has1 mRNA expression were quantified via qPCR.
TRPA1 was expressed in cultured VFFs and the lamina propria. TGF-β1 administration significantly increased Trpa1 compared to control. AITC alone did not alter Smad3, Smad7, Acta2, or ECM related genes. However, the combination of AITC and TGF-β1 significantly increased Smad3 and decreased Smad7 and Acta2 compared to TGF-β1 alone; A-967079 significantly reduced this response.
VFFs expressed TRPA1, and the activation of TRPA1 regulated TGF-β1/Smad signaling in VFFs. These findings provide preliminary insights into potential anti-fibrotic mechanisms of TRPA1 activation through TGF-β1/Smad signaling in VFFs.
NA Laryngoscope, 134:4593-4598, 2024.
声带瘢痕仍然是一个治疗挑战。声带成纤维细胞（VFFs）分泌细胞外基质（ECM），转化生长因子-β1（TGF-β1）介导的成纤维细胞向肌成纤维细胞的分化是纤维化发展的核心。瞬时受体电位（TRP）通道超家族是一组非选择性阳离子通道，TRP 锚蛋白 1（TRPA1）通道的激活已被证明通过 TGF-β1/Smad 信号通路在各种器官中具有抗纤维化作用。本研究旨在阐明 TRPA1 的表达以及 TRPA1 激活对 VFFs 中 TGF-β1/Smad 信号通路的影响。
从 10 周龄雄性 Sprague-Dawley 大鼠分离声带，建立原代 VFFs。通过免疫染色检测 VFFs 和固有层中的 TRPA1。用丙烯基异硫氰酸酯（AITC，TRP 通道激动剂，10 μM）±TGF-β1（10ng/ml）±A-967079（选择性 TRPA1 通道拮抗剂，5.0×10 μM）处理 VFFs 4 或 24 小时。通过 qPCR 定量 Trpa1、Smad3、Smad7、Col1a1、Acta2 和 Has1 的 mRNA 表达。
TRPA1 在培养的 VFFs 和固有层中表达。与对照组相比，TGF-β1 给药显著增加了 Trpa1。单独使用 AITC 不会改变 Smad3、Smad7、Acta2 或 ECM 相关基因。然而，与 TGF-β1 单独给药相比，AITC 和 TGF-β1 的组合显著增加了 Smad3，降低了 Smad7 和 Acta2；A-967079 显著降低了这种反应。
VFFs 表达 TRPA1，TRPA1 的激活调节 VFFs 中的 TGF-β1/Smad 信号通路。这些发现为通过 TGF-β1/Smad 信号通路在 VFFs 中激活 TRPA1 潜在的抗纤维化机制提供了初步见解。
无。喉镜，134：4593-4598，2024。
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