House dust mite and Th2 cytokine-mediated epithelial barrier dysfunction attenuation by KL001 in 16-HBE cells.
House dust mite and Th2 cytokine-mediated epithelial barrier dysfunction attenuation by KL001 in 16-HBE cells.
复制标题
KL001 在 16-HBE 细胞中减弱屋尘螨和 Th2 细胞因子介导的上皮屏障功能障碍。
DOI：
10.1080/21688370.2023.2203841
复制
发表时间：
2024
期刊：
影响因子：
3.1
通讯作者：
Sundar,IsaacKirubakaran
中科院分区：
文献类型：
--
作者：
Duraisamy,SanthoshKumar;Srinivasan,Ashokkumar;Sundar,IsaacKirubakaran
屋尘螨(HDM)是一种常见的空气变应原，可破坏呼吸道上皮屏障，导致免疫反应失调，导致哮喘等过敏性肺部疾病。隐花色素是一种生物钟基因，在新陈代谢和免疫反应中起着重要的调节作用。目前尚不清楚使用KL001稳定CRY是否能减轻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型细胞因子(IL-4或IL-13)介导的上皮屏障功能改变的影响。用XCELLigence实时细胞分析仪检测hDM和Th2型细胞因子诱导的跨上皮细胞电阻的变化，用免疫组织化学和共聚焦显微镜观察黏附连接复合体(AJC：E-钙粘素和β-连环蛋白)和紧密连接蛋白(TJP：occludin和阻断带-1)的非定位化。用实时定量聚合酶链式反应(qRT-PCR)和Western blotting分别检测上皮屏障功能基因和核心时钟基因的表达变化和蛋白丰度。HDM和Th2细胞因子治疗显著降低了与所选上皮屏障功能和生物钟基因的基因表达和蛋白丰度改变相关的TEER。然而，KL001的预治疗可在12-24小时内减轻HDM和Th2细胞因子诱导的上皮屏障功能障碍。KL001可减弱HDM和Th2型细胞因子诱导的AJP和TJP(CDH1、Ocln和Zo1)和核心时钟基因(Clock、Arnt1/BMal1、Cry1/2、Per1/2、Nr1d1/Rev-erbα和Nfil3)的定位和基因表达的改变。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。
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 (Cdh1, Ocln,andZo1) and core clock genes (Clock, Arntl/Bmal1, Cry1/2, Per1/2, Nr1d1/Rev-erbα,andNfil3). We demonstrate, for the first time, the protective role of KL001 in HDM and Th2 cytokine-mediated epithelial barrier dysfunction.