Ozawa Satomi, Mukudai Shigeyuki, Sugiyama Yoichiro, Branski Ryan C, Hirano Shigeru
Department of Otolaryngology-Head and Neck Surgery, Kyoto Prefectural University of Medicine, Kyoto, Japan.
Department of Rehabilitation Medicine, NYU Grossman School of Medicine, New York, New York, U.S.A.
Laryngoscope. 2021 Oct;131(10):2285-2291. doi: 10.1002/lary.29355. Epub 2020 Dec 30.
OBJECTIVES/HYPOTHESIS: Vocal fold fibrosis remains a significant clinical challenge. Estrogens, steroid hormones predominantly responsible for secondary sexual characteristics in women, have been shown to alter wound healing and limit fibrosis, but the effects on vocal fold fibrosis are unknown. We sought to elucidate the expression of estrogen receptors and the effects of estrogens on TGF-β1 signaling in rat vocal fold fibroblasts (VFFs).
In vitro.
VFFs were isolated from 10-week-old, male Sprague-Dawley rats, and estrogen receptor alpha (ERα) and G protein-coupled receptor 30 (GPR30) were examined via immunostaining and quantitative polymerase chain reaction (qPCR). VFFs were treated with estradiol (E2, 10 , 10 or 10 M) ± transforming growth factor beta 1 (TGF-β1, 10 ng/mL). ICI 182,780 (ICI, 10 M) or G36 (10 M) were employed as antagonists of ERα or GPR30, respectively. qPCR was employed to determine estrogen receptor-mediated effects of E2 on genes related to fibrosis.
ERα and GPR30 were expressed in VFFs at both the protein and the mRNA levels. E2 (10 M) did not alter Smad3, Smad7, Acta2 mRNA, or extracellular matrix related genes. However, the combination of E2 (10 M) and TGF-β1 significantly increased Smad7 (P = .03) and decreased Col1a1 (P = .04) compared to TGF-β1 alone; this response was negated by the combination of ICI and G36 (P = .009).
E2 regulated TGF-β1/Smad signaling via estrogen receptors in VFFs. These findings provide insight into potential mechanisms of estrogens on vocal fold injury with the goal of enhanced therapeutics for vocal fold fibrosis.
NA Laryngoscope, 131:2285-2291, 2021.
目的/假设：声带纤维化仍然是一个重大的临床挑战。雌激素是主要负责女性第二性征的类固醇激素，已被证明可改变伤口愈合并限制纤维化，但对声带纤维化的影响尚不清楚。我们试图阐明雌激素受体在大鼠声带成纤维细胞（VFFs）中的表达以及雌激素对转化生长因子-β1（TGF-β1）信号传导的影响。
体外研究。
从10周龄雄性Sprague-Dawley大鼠中分离出VFFs，通过免疫染色和定量聚合酶链反应（qPCR）检测雌激素受体α（ERα）和G蛋白偶联受体30（GPR30）。VFFs用雌二醇（E2，10⁻⁹、10⁻⁸或10⁻⁷M）±转化生长因子-β1（TGF-β1，10 ng/mL）处理。ICI 182,780（ICI，10⁻⁶M）或G36（10⁻⁶M）分别用作ERα或GPR30的拮抗剂。采用qPCR确定E2对纤维化相关基因的雌激素受体介导的影响。
ERα和GPR30在VFFs中均在蛋白和mRNA水平表达。E2（10⁻⁷M）未改变Smad3、Smad7、Acta2 mRNA或细胞外基质相关基因。然而，与单独使用TGF-β1相比，E2（10⁻⁷M）和TGF-β1的组合显著增加了Smad7（P = 0.03）并降低了Col1a1（P = 0.04）；ICI和G36的组合消除了这种反应（P = 0.009）。
E2通过VFFs中的雌激素受体调节TGF-β1/Smad信号传导。这些发现为雌激素对声带损伤的潜在机制提供了见解，目标是增强声带纤维化的治疗效果。
NA 《喉镜》，131:2285 - 2291，2021年。
