Centre for Tissue Engineering and Regenerative Medicine, Faculty of Medicine, Universiti Kebangsaan Malaysia, Cheras, Kuala Lumpur 56000, Malaysia.
Graduate School of Medicine, KPJ Healthcare University College, Kota Seriemas, Nilai 71800, Malaysia.
Int J Mol Sci. 2023 Feb 16;24(4):3974. doi: 10.3390/ijms24043974.
Hydroxytyrosol (HT) is an olive polyphenol with anti-inflammatory and antioxidant properties. This study aimed to investigate the effect of HT treatment on epithelial-mesenchymal transition (EMT) in primary human respiratory epithelial cells (RECs) isolated from human nasal turbinate. HT dose-response study and growth kinetic study on RECs was performed. Several approaches on HT treatment and TGFβ1 induction with varying durations and methods was studied. RECs morphology and migration ability were evaluated. Vimentin and E-cadherin immunofluorescence staining and Western blotting [E-cadherin, vimentin, SNAIL/SLUG, AKT, phosphorylated (p)AKT, SMAD2/3 and pSMAD2/3] were performed after 72-h treatment. In silico analysis (molecular docking) of HT was performed to evaluate the potential of HT to bind with the TGFβ receptor. The viability of the HT-treated RECs was concentration-dependent, where the median effective concentration (EC) was 19.04 μg/mL. Testing of the effects of 1 and 10 µg/mL HT revealed that HT suppressed expression of the protein markers vimentin and SNAIL/SLUG while preserving E-cadherin protein expression. Supplementation with HT protected against SMAD and AKT pathway activation in the TGFβ1-induced RECs. Furthermore, HT demonstrated the potential to bind with ALK5 (a TGFβ receptor component) in comparison to oleuropein. TGFβ1-induced EMT in RECs and HT exerted a positive effect in modulating the effects of EMT.
羟基酪醇 (HT) 是一种具有抗炎和抗氧化特性的橄榄多酚。本研究旨在探讨 HT 处理对人鼻甲骨分离的原代人呼吸道上皮细胞 (RECs) 上皮-间充质转化 (EMT) 的影响。对 HT 处理的 RECs 进行了剂量反应研究和生长动力学研究。研究了不同时间和方法的 HT 处理和 TGFβ1 诱导的几种方法。评估了 RECs 的形态和迁移能力。进行了波形蛋白和 E-钙黏蛋白免疫荧光染色和 Western blot [E-钙黏蛋白、波形蛋白、SNAIL/SLUG、AKT、磷酸化 (p)AKT、SMAD2/3 和 pSMAD2/3]，处理 72 小时后。对 HT 进行了计算机模拟分析（分子对接），以评估 HT 与 TGFβ 受体结合的潜力。HT 处理的 RECs 的活力呈浓度依赖性，其中中效浓度 (EC) 为 19.04μg/ml。测试 1 和 10μg/ml HT 的效果表明，HT 抑制了蛋白标志物波形蛋白和 SNAIL/SLUG 的表达，同时保留了 E-钙黏蛋白的蛋白表达。HT 补充剂可防止 TGFβ1 诱导的 RECs 中 SMAD 和 AKT 通路的激活。此外，与橄榄苦苷相比，HT 显示出与 ALK5（TGFβ 受体成分）结合的潜力。TGFβ1 诱导的 EMT 在 RECs 中，HT 对调节 EMT 效应具有积极作用。
