PM2.5 Aggravated OVA-Induced Epithelial Tight Junction Disruption Through Fas Associated via Death Domain-Dependent Apoptosis in Asthmatic Mice

J Asthma Allergy. 2021 Nov 20;14:1411-1423. doi: 10.2147/JAA.S335590. eCollection 2021.

Abstract

Background: Exposure to air pollutants cause exacerbation of asthma, but the experimental evidence and the mechanisms still need to be collected and addressed.

Methods: Asthma model was constructed by ovalbumin (OVA) combined with or without airborne fine particulate matter 2.5 (PM2.5) exposure. Lung sections were stained by hematoxylin-eosin staining (H&E) and Masson's trichrome. RNA-seq and gene set enrichment analysis (GSEA) was performed to identify the key pathway. TdT mediated dUTP Nick End Labeling (TUNEL) assay, real-time qPCR, Western blot, immunofluorescence and lentivirus transfection were applied for mechanism discovery.

Results: In this study, we found PM2.5 aggravated airway inflammation in OVA-induced asthmatic mice. RNA-seq analysis also showed that epithelial mesenchymal transition (EMT) was enhanced in OVA-induced mice exposed to PM2.5 compared with that in OVA-induced mice. In the meantime, we observed that apoptosis was significantly increased in asthmatic mice exposed to PM2.5 by using GSEA analysis, which was validated by TUNEL assay. By using bioinformatic analysis, Fas associated via death domain (FADD), a new actor in innate immunity and inflammation, was identified to be related to apoptosis, EMT and tight junction. Furthermore, we found that the transcript and protein levels of tight junction markers, E-cadherin, zonula occludens (ZO)-1 and Occludin, were decreased after PM2.5 exposure in vivo and in vitro by using RT-qPCR and immunofluorescence, with the increased expression of FADD. Moreover, down-regulation of FADD attenuated PM2.5-induced apoptosis and tight junction disruption in human airway epithelial cells.

Conclusion: Taken together, we demonstrated that PM2.5 aggravated epithelial tight junction disruption through apoptosis mediated by up-regulation of FADD in OVA-induced model.

Keywords: FADD; PM2.5; apoptosis; asthma; epithelial tight junction disruption.

Grant support

This work was supported by the National Natural Science Foundation of China (81970026, 82000029), Health Commission of Sichuan Province (19ZD002, 20PJ208), Science and Technology Department of Sichuan Province (2018JY0380), Health Commission of Chengdu (2021011, 2021021, 2021040), Chengdu High-level Key Clinical Specialty Construction Project (ZX20201202020), Sichuan Province Administration of Traditional Chinese Medicine (2020JC0118), Chengdu Science and Technology Bureau (2021-YF09-00102-SN, 2020-YF05-00003-SN), the program for combination of Medical and Engineering of Southwest Jiaotong University (2682021ZTPY007, 2682020ZT8).