Transcriptomic profiling of human corneal epithelial cells exposed to airborne fine particulate matter (PM2.5)
Transcriptomic profiling of human corneal epithelial cells exposed to airborne fine particulate matter (PM2.5)
复制标题
暴露于空气细颗粒物 (PM2.5) 的人角膜上皮细胞的转录组分析
DOI：
10.1016/j.jtos.2020.06.003
复制
发表时间：
2020-10-01
期刊：
影响因子：
6.4
通讯作者：
Fu, Qiuli
中科院分区：
文献类型：
--
作者：
Lyu, Danni;Chen, Zhijian;Fu, Qiuli
目的：探讨PM2.5诱导的人角膜上皮细胞功能障碍的分子机制，以及纤溶酶原激活物抑制物-2(PAI-2)在PM2.5诱导的体内外自噬中的作用。方法：采用RNA-Seq技术鉴定PM2.5暴露的人角膜上皮细胞与未暴露的角膜上皮细胞的差异表达基因，并通过实时定量聚合酶链式反应(qRT-PCR)进行验证。在PM2.5暴露的大鼠模型中检测角膜荧光素染色和泪液分泌。通过免疫印迹、免疫荧光染色和/或qRT-PCR检测PM2.5暴露或未暴露的内皮细胞和大鼠角膜中PAI-2和自噬相关标志物的表达。PAI-2基因敲除人脐静脉内皮细胞，研究PAI-2‘S在PM2.5诱导的人脐静脉内皮细胞自噬中的作用。PM2.5暴露显著改变了一些与增殖、炎症和芳烃刺激相关的基因的表达。PAI-2在PM2.5暴露的人脐静脉内皮细胞中表达上调，与自噬相关标记物LC3B II和BECN1在不同暴露时间的变化趋势相似。此外，PAI-2基因敲除可显著抑制PM2.5诱导的内皮细胞中LC3B和BECN1的表达。PM2.5暴露组大鼠角膜荧光素染色明显增强，泪液分泌明显减少。PAI-2在PM2.5暴露的大鼠角膜中的表达也增加，并上调了几个自噬相关标志物的表达。结论：本研究发现PM2.5暴露的HCECs中数百个基因的表达发生了变化，这表明PM2.5对角膜健康具有重要意义。PAI-2参与PM2.5诱导的内皮细胞自噬以及大鼠角膜的自噬，提示PAI-2可能成为PM2.5相关眼表疾病临床治疗的潜在靶点。
Purpose: To explore the molecular mechanisms of PM2.5-induced dysfunction in human corneal epithelial cells (HCECs) and the potential role of the plasminogen activator inhibitor type-2 (PAI-2) in PM2.5-induced autophagy in vitro and in vivo.Methods: RNA-Seq was performed to identify the differentially expressed genes (DEGs) in PM2.5-exposed HCECs compared to unexposed condition, followed by validation via real-time PCR (qRT-PCR). Corneal fluorescein staining and tear secretion were assessed in the PM2.5-exposed rat model. The expression of PAI-2 and autophagy-related markers were examined via immunoblotting, immunofluorescence staining and/or qRT-PCR in PM2.5-exposed or unexposed HCECs and rat corneas. PAI-2-knockdown HCECs were generated to study PAI-2's role in the PM2.5-induced autophagy in HCECs.Results: A total of 434 DEGs-240 up-regulated and 194 down-regulated-were identified in PM2.5-exposed HCECs rather than unexposed HCECs. The expression of a few genes related to proliferation, inflammation, and aryl hydrocarbon stimulation were significantly altered by PM2.5 exposure. PAI-2 expression was up-regulated in PM2.5-exposed HCECs, sharing a similar fluctuation trend with autophagy-related markers LC3B II and BECN1 according to various exposure periods. Moreover, PAI-2 knockdown significantly suppressed the expression of LC3B and BECN1 in PM2.5-exposed HCECs. The corneal fluorescein staining was enhanced and tear secretion was significantly reduced in PM2.5-exposed rat eyes. PAI-2 expression was also increased in PM2.5-exposed rat corneas, together with the up-regulation of several autophagy-related markers.Conclusion: The present study identified the altered expression of hundreds of genes in PM2.5-exposed HCECs, which suggests the importance of PM2.5 for cornea health. The involvement of PAI-2 was discovered in the PM2.5-induced autophagy in HCECs as well as likely in rat corneas, which implied that PAI-2 may become a potential target of clinical treatment of PM2.5-associated ocular surface diseases.