Transcriptomic profiling of human corneal epithelial cells exposed to airborne fine particulate matter (PM2.5).,The Ocular Surface

当前位置： X-MOL 学术 › Ocul. Surf. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Transcriptomic profiling of human corneal epithelial cells exposed to airborne fine particulate matter (PM2.5).
The Ocular Surface ( IF 5.9 ) Pub Date : 2020-06-19 , DOI: 10.1016/j.jtos.2020.06.003
Danni Lyu ₁ , Zhijian Chen ₂ , Siham Almansoob ₁ , Hui Chen ₁ , Yang Ye ₁ , Fan Song ₁ , Lifang Zhang ₁ , Zhenwei Qin ₁ , Qiaomei Tang ₁ , Houfa Yin ₁ , Wen Xu ₁ , Ke Yao ₁ , Qiuli Fu ₁

Affiliation

Purpose

To explore the molecular mechanisms of PM_2.5-induced dysfunction in human corneal epithelial cells (HCECs) and the potential role of the plasminogen activator inhibitor type-2 (PAI-2) in PM_2.5-induced autophagy in vitro and in vivo.

Methods

RNA-Seq was performed to identify the differentially expressed genes (DEGs) in PM_2.5-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 PM_2.5-exposed rat model. The expression of PAI-2 and autophagy-related markers were examined via immunoblotting, immunofluorescence staining and/or qRT-PCR in PM_2.5-exposed or unexposed HCECs and rat corneas. PAI-2-knockdown HCECs were generated to study PAI-2's role in the PM_2.5-induced autophagy in HCECs.

Results

A total of 434 DEGs—240 up-regulated and 194 down-regulated—were identified in PM_2.5-exposed HCECs rather than unexposed HCECs. The expression of a few genes related to proliferation, inflammation, and aryl hydrocarbon stimulation were significantly altered by PM_2.5 exposure. PAI-2 expression was up-regulated in PM_2.5-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 PM_2.5-exposed HCECs. The corneal fluorescein staining was enhanced and tear secretion was significantly reduced in PM_2.5-exposed rat eyes. PAI-2 expression was also increased in PM_2.5-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 PM_2.5-exposed HCECs, which suggests the importance of PM_2.5 for cornea health. The involvement of PAI-2 was discovered in the PM_2.5-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 PM_2.5-associated ocular surface diseases.

中文翻译：

暴露于空气传播的细颗粒物（PM2.5）的人角膜上皮细胞的转录组分析。

目的

探讨PM _2.5诱导的人角膜上皮细胞（HCEC）功能障碍的分子机制，以及纤溶酶原激活物抑制剂2型（PAI-2）在PM _2.5诱导的自噬体内外的潜在作用。

方法

进行RNA-Seq以鉴定暴露于PM _2.5的HCEC中与未暴露条件相比的差异表达基因（DEG），然后通过实时PCR（qRT-PCR）进行验证。在暴露于PM _2.5的大鼠模型中评估了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PM _2.5暴露或未暴露的HCEC和大鼠角膜中PAI-2和自噬相关标记的表达。生成了PAI-2-nockdown HCEC，以研究PAI-2在HCEC中PM _2.5诱导的自噬中的作用。

结果

在PM _2.5暴露的HCEC中而不是未暴露的HCEC中总共确定了434个DEG（240个上调和194个下调）。与PM _2.5接触显着改变了与增殖，炎症和芳基烃刺激相关的一些基因的表达。在暴露于PM _2.5的HCEC中，PAI-2表达上调，并根据不同的暴露时间与自噬相关标记LC3B II和BECN1共享相似的波动趋势。而且，PAI-2敲低显着抑制了暴露于PM _2.5的HCEC中LC3B和BECN1的表达。在PM _2.5中，角膜荧光素染色增强，泪液分泌明显减少暴露的老鼠眼睛。在暴露于PM _2.5的大鼠角膜中，PAI-2的表达也增加了，同时上调了几种自噬相关标记。