鉴定和验证糖尿病视网膜病变中的自噬相关基因。
Identification and Validation of Autophagy-Related Genes in Diabetic Retinopathy.
机构信息
Eye Center of Xiangya Hospital, Central South University, Changsha, China.
Hunan Key Laboratory of Opthalmology, Central South University, Changsha, China.
出版信息
Front Endocrinol (Lausanne). 2022 Apr 29;13:867600. doi: 10.3389/fendo.2022.867600. eCollection 2022.
BACKGROUND
Diabetic retinopathy (DR) is one of the most common microvascular complications of diabetes, which is associated with damage of blood-retinal barrier and ischemia of retinal vasculature. It devastates visual acuity due to leakage of retinal vessels and aberrant pathological angiogenesis in diabetic patients. The etiology of DR is complex, accumulated studies have shown that autophagy plays an important role in the pathogenesis of DR, but its specific mechanism needs to be further studied.
METHODS
This study chose the online Gene Expression Omnibus (GEO) microarray expression profiling dataset GSE146615 to carry on the research. Autophagy-related genes that were potentially differentially expressed in DR were screened by R software. Then, the differentially expressed autophagy-related genes were analyzed by correlation analysis, tissue-specific gene expression, gene-ontology (GO) enrichment analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis and protein-protein interaction (PPI) network analysis. Finally, retinal pigment epithelial cell line (ARPE-19) incubated with high glucose (HG) was used to mimic the DR model, and the mRNA level of key genes was verified by quantitative real-time polymerase chain reaction (qRT-PCR) .
RESULTS
A total of 23 differentially expressed autophagy-related genes (9 up-regulated genes and 14 down-regulated genes) were identified by differential expression analysis. The analysis of tissue-specific gene expression showed that these differentially expressed autophagy-related genes were enriched in the retina. GO and KEGG enrichment analysis showed that differentially expressed autophagy-related genes were significantly enriched in autophagy-related pathways such as regulation of autophagy and macroautophagy. Then 10 hub genes were identified by PPI network analysis and construction of key modules. Finally, qRT-PCR confirmed that the expression of MAPK3 in the DR model was consistent with the results of bioinformatics analysis of mRNA chip.
CONCLUSION
Through bioinformatics analysis, we identified 23 potential DR autophagy-related genes, among which the down-regulated expression of MAPK3 may affect the occurrence and development of DR by regulating autophagy. It provides a novel insight into the pathogenesis of DR.
背景
糖尿病视网膜病变（DR）是糖尿病最常见的微血管并发症之一，与血视网膜屏障损伤和视网膜血管缺血有关。它会破坏糖尿病患者的视力，原因是视网膜血管渗漏和异常病理性血管生成。DR 的病因复杂，已有大量研究表明自噬在 DR 的发病机制中起重要作用，但具体机制仍需进一步研究。
方法
本研究选择在线基因表达综合数据库（GEO）微阵列表达谱数据集 GSE146615 进行研究。通过 R 软件筛选 DR 中潜在差异表达的自噬相关基因。然后，通过相关性分析、组织特异性基因表达分析、基因本体论（GO）富集分析、京都基因与基因组百科全书（KEGG）通路富集分析和蛋白质-蛋白质相互作用（PPI）网络分析对差异表达的自噬相关基因进行分析。最后，用高糖（HG）孵育人视网膜色素上皮细胞系（ARPE-19）模拟 DR 模型，并通过实时定量聚合酶链反应（qRT-PCR）验证关键基因的 mRNA 水平。
结果
通过差异表达分析，共鉴定出 23 个差异表达的自噬相关基因（9 个上调基因和 14 个下调基因）。组织特异性基因表达分析表明，这些差异表达的自噬相关基因在视网膜中富集。GO 和 KEGG 富集分析表明，差异表达的自噬相关基因显著富集在自噬相关途径，如自噬调节和巨自噬。然后通过 PPI 网络分析和关键模块的构建，鉴定出 10 个枢纽基因。最后，qRT-PCR 验证了 DR 模型中 MAPK3 的表达与 mRNA 芯片的生物信息学分析结果一致。
结论
通过生物信息学分析，我们确定了 23 个潜在的 DR 自噬相关基因，其中 MAPK3 的下调表达可能通过调节自噬影响 DR 的发生和发展。这为 DR 的发病机制提供了新的见解。
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