The Effect of Zerumbone on Tight Junctional Proteins Localization in House Dust Mite-Induced Airway Epithelial Barrier Disruption

Mohd Sebri, Nor Farhanah (2020) The Effect of Zerumbone on Tight Junctional Proteins Localization in House Dust Mite-Induced Airway Epithelial Barrier Disruption. [Project Paper] (Submitted)

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Abstract

Human airway systems are lined by epithelial cells that are attached to each other by junctional proteins. These junctional proteins act as a physical barrier that limits the diffusion of substances across the intercellular spaces. Allergen such as house dust mite (HDM) can disrupt the epithelial barrier via junctional disassembly resulting in respiratory diseases such as asthma. Zerumbone is a monocyclic sesquiterpene compound found in the rhizome of wild ginger, Zingiber zerumbet (L) Smith. Previous study has shown that Zerumbone possesses anti-asthmatic effect by modulating Th1/Th2 cytokines. Our research group also recently found that Zerumbone was able to attenuate the loss of junctional integrity and increase the paracellular permeability in HDM-induced epithelial barrier dysfunction. However, Zerumbone was not able to prevent the cleavage/degradation of ZO-1 and occludin. Objective: This study aims to investigate the effect of Zerumbone on tight junctional proteins localization in HDM-induced airway epithelial barrier disruption. Methodology: An immortalized human bronchial epithelial cell line (16HBE14o-) was used as a cellular model of the airway epithelial barrier. 16HBE14o- cells were treated with increasing concentrations of Zerumbone (6.25 μM-200 μM) for 24 hours and the cell viability was assessed using MTT assay. The cells were then co-treated with 100 μg/mL HDM and non-cytotoxic concentrations of Zerumbone (6.25, 12.5 and 25 μM) for 24 hours, followed by immunofluorescence (IF) assay to investigate the localization of tight junctional proteins, ZO-1 and occludin. The gap area of both junctional proteins was measured by analyzing the images using ImageJ software. Results: No significant cell death was observed at a concentration of 25µM and below. Zerumbone was shown to preserve the localization of ZO-1 and occludin at the periphery of 16HBE14o- cells. This was confirmed by a significant reduction of the gap area in Zerumbone treatment group for ZO-1 (12.5 and 25 μM, p<0.01 and p<0.001) and occludin (12.5 and 25 μM, p<0.05 and p<0.001) respectively. Conclusion: Zerumbone possesses protective effects on HDM-induced airway epithelial barrier disruption by maintaining the localization of tight junctional proteins (ZO-1 and occludin).

Item Type:	Project Paper
Faculty:	Faculty of Medicine and Health Science
Depositing User:	Ms. Nor Safa'aton Saidin
Date Deposited:	23 Aug 2023 00:47
Last Modified:	23 Aug 2023 00:47
URI:	http://psaspb.upm.edu.my/id/eprint/1288

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