Zhou Ziyu, Gao Zhaolin, Yan Weitao, Zhang Yun, Huang Jufang, Xiong Kun
Department of Anatomy and Neurobiology, School of Basic Medical Sciences, Central South University, Changsha, China.
School of Life Sciences, Central South University, Changsha, China.
Ann Transl Med. 2021 Apr;9(7):526. doi: 10.21037/atm-20-6154.
Primary open-angle glaucoma (POAG), as one of the leading reasons for blindness, is mainly due to trabecular meshwork (TM) dysfunction. Bioinformatics analysis was used to find related genes involved in TM oxidative stress, which is a major cause of TM fibrosis.
A total of three datasets from the Gene Expression Omnibus (GEO) database were used to identify differentially expressed genes (DEGs). Gene expression relationships were enriched by the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and Gene Ontology (GO) pathways. The interaction network was listed by the protein-protein interaction (PPI) network. The expression of adenosine A3 receptor (ADORA3) was validated in POAG tissue and human trabecular meshwork cells (HTMCs) by western blot (WB) and reverse transcription polymerase chain reaction (RT-PCR). Additionally, WB and RT-PCR were used to measure oxidative stress injury relative protein and gene expression, respectively, such as fibronectin (FN), collagen-I (Col-I), and α-smooth muscle actin (α-SMA). Cell migration function and vitality were tested via transwell migration assay and Cell Counting Kit-8 (CCK-8). The cell vitality was measured using CCK-8.
A total of 61 significant DEGs among the three data sources were analyzed. Among all three different datasets, two significant DEGs [ADORA3 and DNA damage-inducible transcript 4 protein (DDIT4)] were identified. The dataset ADORA3 was selected for further analysis. In the POAG TM tissue, ADORA3 was overexpressed at transcriptional and post-transcriptional levels. Overexpression of ADORA3 reduced TMC viability and migration but upregulated the extracellular matrix (ECM) proteins (FN, Col-I, and α-SMA) expression. It was found that ADORA3 can exacerbate oxidative stress injury in normal TMCs. These results indicated that ADORA3 might play an essential role in the occurrence and progression of POAG.
A total of 61 novel common DEGs identified are related to the development and prognosis of POAG. In the POAG, ADORA3 was verified as overexpressed; therefore, it may be associated with an oxidative stress injury in TMCs.
原发性开角型青光眼（POAG）是导致失明的主要原因之一，主要归因于小梁网（TM）功能障碍。采用生物信息学分析来寻找参与TM氧化应激的相关基因，TM氧化应激是TM纤维化的主要原因。
使用来自基因表达综合数据库（GEO）的总共三个数据集来鉴定差异表达基因（DEG）。通过京都基因与基因组百科全书（KEGG）通路和基因本体论（GO）通路来富集基因表达关系。通过蛋白质-蛋白质相互作用（PPI）网络列出相互作用网络。通过蛋白质印迹法（WB）和逆转录聚合酶链反应（RT-PCR）在POAG组织和人小梁网细胞（HTMC）中验证腺苷A3受体（ADORA3）的表达。此外，分别使用WB和RT-PCR来测量氧化应激损伤相关蛋白和基因的表达，如纤连蛋白（FN）、I型胶原（Col-I）和α-平滑肌肌动蛋白（α-SMA）。通过Transwell迁移试验和细胞计数试剂盒-8（CCK-8）检测细胞迁移功能和活力。使用CCK-8测量细胞活力。
分析了三个数据源中的总共61个显著DEG。在所有三个不同的数据集中，鉴定出两个显著的DEG[ADORA3和DNA损伤诱导转录4蛋白（DDIT4）]。选择数据集ADORA3进行进一步分析。在POAG TM组织中，ADORA3在转录和转录后水平均过表达。ADORA3的过表达降低了TMC的活力和迁移能力，但上调了细胞外基质（ECM）蛋白（FN、Col-I和α-SMA）的表达。发现ADORA3可加重正常TMC中的氧化应激损伤。这些结果表明ADORA3可能在POAG的发生和发展中起重要作用。
总共鉴定出的61个新的常见DEG与POAG的发生和预后相关。在POAG中，ADORA3被证实过表达；因此，它可能与TMC中的氧化应激损伤有关。
