Chen Li-Da, Huang Jie-Feng, Lin Xue-Jun, Huang Ya-Ping, Xu Qiao-Zhen, Chen Gong-Ping, Lin Qi-Chang
Department of Respiratory and Critical Care Medicine, Zhangzhou Affiliated Hospital of Fujian Medical University, Zhangzhou, China.
Department of Respiratory and Critical Care Medicine, the First Affiliated Hospital of Fujian Medical University, Fuzhou, China.
Front Physiol. 2022 Sep 14;13:972407. doi: 10.3389/fphys.2022.972407. eCollection 2022.
Intermittent hypoxia (IH) is a prominent feature of obstructive sleep apnea (OSA) which is increasingly recognized as a key risk factor for liver injury. Circular RNAs (circRNAs) has been suggested to act as a regulator of multiple biological processes. However, there is no study evaluating circRNAs alterations and potential role of circRNAs in OSA-related liver injury. The present study aimed to investigate circRNA expression profiles model of IH-induced liver injury, as well as potential functional characterization of the differentially expressed circRNAs (DE circRNAs). BRL-3A cells were exposed to IH or normoxia. Cell apoptosis and cell viability were evaluated using flow cytometry and cell counting kit-8, respectively. The expression profile of circRNAs was depicted by circRNA sequencing. The selected circRNAs were verified by quantitative real-time PCR (qRT-PCR). Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and Gene Ontology (GO) analyses were employed to predict DE circRNAs functions. The circRNA-miRNA-mRNA regulatory network was constructed. IH treatment caused cell injury in BRL-3A cells. 98 circRNAs were identified as being dysregulated in IH-treated BRL-3A cells. Among them, 58 were up-regulated and 40 were down-regulated. Go and KEGG analyses suggested that the DE circRNAs were predominantly enriched in the biological process such as positive regulation of NF-kappaB transcription factor activity and pathways such as circadian entrainment, Wnt signaling pathway, MAPK signaling pathway, and protein export. 3 up-regulated circRNAs and 3 down-regulated circRNAs with high number of back-splicing sites were chosen for qRT-PCR validation and were consistent with the sequencing data. CircRNA1056 and circRNA805 were predicted to interact with microRNAs that might thereby regulate downstream genes. The study characterized a profile of dysregulated circRNAs in IH-induced BRL-3A cell injury. DE circRNAs may play vital roles in the pathophysiology of IH-induced liver injury. Our findings provide preliminary support for further research in mechanisms and a new theory for the pathogenesis of OSA-related liver injury.
间歇性缺氧（IH）是阻塞性睡眠呼吸暂停（OSA）的一个显著特征，OSA日益被认为是肝损伤的一个关键危险因素。环状RNA（circRNAs）已被认为可作为多种生物学过程的调节因子。然而，尚无研究评估circRNAs的变化及其在OSA相关肝损伤中的潜在作用。本研究旨在探究IH诱导的肝损伤的circRNA表达谱模型，以及差异表达的circRNAs（DE circRNAs）的潜在功能特征。将BRL-3A细胞暴露于IH或常氧环境中。分别使用流式细胞术和细胞计数试剂盒-8评估细胞凋亡和细胞活力。通过circRNA测序描绘circRNAs的表达谱。通过定量实时PCR（qRT-PCR）验证所选的circRNAs。采用京都基因与基因组百科全书（KEGG）通路和基因本体论（GO）分析来预测DE circRNAs的功能。构建circRNA- miRNA- mRNA调控网络。IH处理导致BRL-3A细胞损伤。在经IH处理的BRL-3A细胞中，有98种circRNAs被鉴定为表达失调。其中，58种上调，40种下调。GO和KEGG分析表明，DE circRNAs主要富集于诸如NF-κB转录因子活性的正调控等生物学过程以及诸如昼夜节律调整、Wnt信号通路、MAPK信号通路和蛋白质输出等通路。选择3种上调的circRNAs和3种下调的circRNAs进行qRT-PCR验证，且结果与测序数据一致。预测circRNA1056和circRNA805与可能由此调节下游基因的微小RNA相互作用。本研究描绘了IH诱导的BRL-3A细胞损伤中失调的circRNAs谱。DE circRNAs可能在IH诱导的肝损伤的病理生理学中发挥重要作用。我们的发现为机制的进一步研究提供了初步支持，并为OSA相关肝损伤的发病机制提供了新的理论依据。
