Division of Pulmonary, Critical Care and Sleep Medicine, Department of Internal Medicine, University of Kansas Medical Center, Kansas City, KS, USA.
Tissue Barriers. 2024 Jan 2;12(1):2203841. doi: 10.1080/21688370.2023.2203841. Epub 2023 Apr 20.
House dust mite (HDM) is a common aeroallergen that can disrupt the airway epithelial barrier leading to dysregulated immune response, resulting in allergic lung diseases such as asthma. Cryptochrome (CRY), a circadian clock gene, plays an important role in the regulation of metabolism, and immune response. It remains unclear whether stabilizing CRY using KL001 can attenuate HDM/Th2 cytokine-induced epithelial barrier dysfunction in 16-HBE cells. We evaluate the effect of KL001 (20 µM) pre-treatment (4 hrs) in HDM/Th2 cytokine (IL-4 or IL-13)-mediated change in epithelial barrier function. HDM and Th2 cytokine-induced changes in transepithelial electrical resistance (TEER) were determined by an xCELLigence real-time cell analyzer and delocalization of adherens junction complex (AJC: E-cadherin and β-catenin) and tight junction proteins (TJP: Occludin and Zonula occludens-1) by immunostaining and confocal microscopy. Finally, quantitative real-time PCR (qRT-PCR) and Western blotting were used to measure altered gene expression and protein abundance of the epithelial barrier function and core clock genes, respectively. HDM and Th2 cytokine treatment significantly decreased TEER associated with altered gene expression and protein abundance of the selected epithelial barrier function and circadian clock genes. However, pre-treatment with KL001 attenuated HDM and Th2 cytokine-induced epithelial barrier dysfunction as early as 12-24 hrs. KL001 pre-treatment showed attenuation of HDM and Th2 cytokine-induced alteration in the localization and gene expression of AJP and TJP ( and ) and core clock genes ( and ). We demonstrate, for the first time, the protective role of KL001 in HDM and Th2 cytokine-mediated epithelial barrier dysfunction.
屋尘螨（HDM）是一种常见的过敏原，它会破坏气道上皮屏障，导致免疫反应失调，从而引发哮喘等过敏性肺部疾病。隐花色素（CRY）是一种生物钟基因，在调节代谢和免疫反应方面发挥着重要作用。目前尚不清楚使用 KL001 稳定 CRY 是否可以减轻 HDM/Th2 细胞因子诱导的 16-HBE 细胞上皮屏障功能障碍。我们评估了 KL001（20 μM）预处理（4 小时）对 HDM/Th2 细胞因子（IL-4 或 IL-13）介导的上皮屏障功能改变的影响。通过 xCELLigence 实时细胞分析仪测定 HDM 和 Th2 细胞因子诱导的跨上皮电阻（TEER）变化，通过免疫染色和共聚焦显微镜测定黏附连接复合物（AJC：E-钙粘蛋白和β-连环蛋白）和紧密连接蛋白（TJP：闭合蛋白和封闭蛋白-1）的定位。最后，使用定量实时 PCR（qRT-PCR）和 Western blot 分别测量上皮屏障功能和核心生物钟基因的改变基因表达和蛋白丰度。HDM 和 Th2 细胞因子处理显著降低了与选定的上皮屏障功能和生物钟基因的改变基因表达和蛋白丰度相关的 TEER。然而，KL001 的预处理早在 12-24 小时即可减轻 HDM 和 Th2 细胞因子诱导的上皮屏障功能障碍。KL001 预处理显示出对 HDM 和 Th2 细胞因子诱导的 AJP 和 TJP（和）和核心生物钟基因（和）的定位和基因表达改变的抑制作用。我们首次证明了 KL001 在 HDM 和 Th2 细胞因子介导的上皮屏障功能障碍中的保护作用。
