Expression Profiles of CircRNA and mRNA in Lacrimal Glands of AQP5-/-Mice With Primary Dry Eye
Expression Profiles of CircRNA and mRNA in Lacrimal Glands of AQP5-/-Mice With Primary Dry Eye
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DOI：
10.3389/fphys.2020.01010
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发表时间：
2020-09-03
影响因子：
4
通讯作者：
Chen, Peng
中科院分区：
文献类型：
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作者：
Liu, Yaning;Di, Guohu;Chen, Peng
目的：本研究旨在鉴定水通道蛋白5(AQP5)基因敲除(AQP5(-/-))小鼠中差异表达的环状RNA(CircRNAs)，并阐明其在干眼症小鼠中的潜在功能。方法：对AQP5(-/-)小鼠进行裂隙灯检查，用荧光素钠染色评价角膜上皮损伤情况。苏木精-伊红染色和透射电子显微镜观察AQP5缺乏对泪腺上皮细胞结构的影响。应用基因芯片技术分析CircRNA和信使RNA的表达谱。用实时定量逆转录-聚合酶链式反应(qRT-PCR)对筛选出的CircRNA进行了验证。基因本体论和京都基因和基因组百科全书(KEGG)途径丰富分析预测亲本基因在泪腺上皮细胞变化中的生物学功能和潜在途径。根据对已鉴定的CircRNA的生物信息学分析，我们预测了一个CircRNA-miRNA-mRNA吞噬小体网络。结果：AQP5(-/-)小鼠自发出现干眼症状，其中AQP5缺陷改变了泪腺上皮细胞的结构。分析表明，与AQP5(+/+)小鼠相比，AQP5(-/-)小鼠泪腺中有30个CircRNA有差异表达(折叠式变化=2.0，p<；0.05)。用qRT-PCR鉴定了9个上调的CircRNAs，通过网络分析鉴定了9个上调的有效CircRNAs、40个改变的microRNAs(MiRNAs)和9个上调的mRNAs。KEGG分析表明，这9个目的基因均在噬菌体中表达。结论：AQP5(-/-)小鼠从出生起就具有稳定的原发干眼症表型。我们鉴定了在AQP5(-/-)和AQP5(+/+)小鼠泪腺中表达不同的CircRNA，预测了一个由吞噬小体组成的CircRNA-miRNA-mRNA网络。CircRNA可能在泪腺上皮细胞的发病机制中起重要作用。因此，CircRNA作为治疗干眼症的潜在药物是合理的。
Purpose:This work aimed to identify differentially expressed circular RNAs (circRNAs) and elucidate their potential function in aquaporin 5 (AQP5) knockout (AQP5(-/-)) mice with the primary dry eye phenotype. Methods:A slit lamp examination was performed on AQP5(-/-)mice to assess corneal epithelial defects using fluorescein sodium staining. Hematoxylin-eosin staining and transmission electron microscopy analysis were performed to identify structural changes in lacrimal gland epithelial cells due to AQP5 deficiency. The expression profiles of circRNA and messenger RNA (mRNA) were determined by a microarray analysis. The selected circRNA was verified by quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR). Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed to predict the biological functions and the potential pathways of parental genes involved in lacrimal gland epithelial cell changes. According to the bioinformatics analysis of identified circRNAs, we predicted a circRNA-miRNA-mRNA network of phagosomes. Results:The AQP5(-/-)mice spontaneously exhibit dry eye symptoms, wherein the AQP5 deficiency changes the structure of lacrimal gland epithelial cells. The analysis revealed that, compared to AQP5(+/+)mice, 30 circRNAs in the lacrimal glands of AQP5(-/-)mice were differentially expressed (fold change >= 2.0,p< 0.05). Nine upregulated circRNAs were identified using qRT-PCR, and nine upregulated validated circRNAs, 40 altered microRNAs (miRNAs), and nine upregulated mRNAs were identified through a network analysis. The KEGG analysis showed that these nine target genes were expressed in phagosomes. Conclusion:The AQP5(-/-)mice have primary and stable dry eye phenotypes from birth. We identified differently expressed circRNAs in the lacrimal glands of AQP5(-/-)and AQP5(+/+)mice, predicting a circRNA-miRNA-mRNA network of phagosomes. CircRNA likely plays an important role in lacrimal gland epithelial cell pathogenesis. Therefore, it is reasonable to use circRNA as a potential therapeutic agent for the treatment of dry eyes.