Adenosine A3 receptor activated in H(2)O(2) oxidative stress of primary open-angle glaucoma.
Adenosine A3 receptor activated in H(2)O(2) oxidative stress of primary open-angle glaucoma.
复制标题
原发性开角型青光眼 H2O2 氧化应激中腺苷 A3 受体激活
DOI：
10.21037/atm-20-6154
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发表时间：
2021-04
影响因子：
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通讯作者：
Xiong K
中科院分区：
文献类型：
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作者：
Zhou Z;Gao Z;Yan W;Zhang Y;Huang J;Xiong K
背景原发性开角型青光眼（primary open-angle glaucoma，POAG）是导致失明的主要原因之一，其主要原因是小梁网（trabecular meshwork，TM）功能障碍。利用生物信息学分析方法寻找TM氧化应激相关基因，该基因是TM纤维化的主要原因。方法利用基因表达综合数据库（GEO）中的3个数据集进行差异表达基因（DEG）分析。基因表达关系通过京都基因和基因组百科全书（KEGG）途径和基因本体（GO）途径来丰富。相互作用网络由蛋白质-蛋白质相互作用（PPI）网络列出。应用Western blot和RT-PCR方法检测POAG组织和人眼小梁细胞中腺苷A3受体（adenosine A3 receptor，ADORA 3）的表达。WB法检测氧化应激损伤相关蛋白的表达，RT-PCR法检测氧化应激损伤相关基因的表达，如纤维连接蛋白（FN）、Ⅰ型胶原（Col-I）和α-平滑肌肌动蛋白（α-SMA）。通过transwell迁移实验和CCK-8试剂盒检测细胞迁移功能和活力。使用CCK-8测量细胞活力。结果在3个数据源中共分析了61个显著的DEG。在所有三个不同的数据集中，鉴定了两个显著的DEG [ADORA 3和DNA损伤诱导转录物4蛋白（DDIT 4）]。选择数据集ADORA 3进行进一步分析。在POAG TM组织中，ADORA 3在转录和转录后水平过表达。ADORA 3的过表达降低了TMC的活力和迁移，但上调了细胞外基质（ECM）蛋白（FN、Col-I和α-SMA）的表达。发现AD 0 RA 3可加重正常TMCs中的氧化应激损伤。提示ADORA 3可能在POAG的发生、发展中起重要作用。结论61例新发现的DEG与POAG的发生、发展及预后有关。在POAG中，ADORA 3被证实为过表达;因此，它可能与TMCs中的氧化应激损伤有关。
Background 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. Methods 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. Results 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. Conclusions 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.
影响因子：
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作者：
He HR;Li YJ;He GH;Qiang H;Zhai YJ;Ma M;Wang YJ;Wang Y;Zheng XW;Dong YL;Lyu J
通讯作者：
Lyu J