PM2.5 Aggravated OVA-Induced Epithelial Tight Junction Disruption Through Fas Associated via Death Domain-Dependent Apoptosis in Asthmatic Mice.
PM2.5 Aggravated OVA-Induced Epithelial Tight Junction Disruption Through Fas Associated via Death Domain-Dependent Apoptosis in Asthmatic Mice.
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DOI：
10.2147/jaa.s335590
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发表时间：
2021
影响因子：
3.2
通讯作者：
Li G
中科院分区：
文献类型：
--
作者：
He X;Zhang L;Hu L;Liu S;Xiong A;Wang J;Xiong Y;Li G
暴露于空气污染物会导致哮喘加重，但实验证据和机制仍需要收集和解决。采用卵清蛋白（OVA）联合或不联合大气细颗粒物2.5（PM2.5）建立哮喘模型。肺切片通过苏木精-伊红染色（H&E）和Masson三色染色。进行RNA-seq和基因集富集分析（GSEA）以鉴定关键途径。采用原位末端标记（TUNEL）、实时定量PCR、Western blot、免疫荧光和慢病毒转染等方法进行机制研究。在本研究中，我们发现PM2.5加重了OVA诱导的哮喘小鼠的气道炎症。RNA-seq分析还显示，与OVA诱导的小鼠相比，暴露于PM2. 5的OVA诱导的小鼠的上皮间质转化（EMT）增强。同时，我们通过GSEA分析观察到PM2.5暴露的哮喘小鼠细胞凋亡显著增加，并通过TUNEL法验证了这一点。通过生物信息学分析，发现Fas通过死亡结构域（FADD）与细胞凋亡、EMT和紧密连接有关，是天然免疫和炎症的一个新的参与者。此外，我们发现，在体内和体外暴露于PM2. 5后，紧密连接标志物，E-cadherin，zonula occludens（ZO）-1和Occludin的转录和蛋白水平降低，FADD的表达增加。此外，下调FADD减弱PM2.5诱导的人气道上皮细胞凋亡和紧密连接破坏。综上所述，我们证明了PM2.5通过在OVA诱导的模型中上调FADD介导的细胞凋亡加重上皮紧密连接的破坏。
Exposure to air pollutants cause exacerbation of asthma, but the experimental evidence and the mechanisms still need to be collected and addressed. 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. 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. Taken together, we demonstrated that PM2.5 aggravated epithelial tight junction disruption through apoptosis mediated by up-regulation of FADD in OVA-induced model.