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目的:筛选增生型糖尿病视网膜病变（DR）患者差异表达基因（DEG ），为增生型DR(PDR）治疗提供新的生物学治疗靶点。方法:基础研究。2020年10月于天津医科大学眼科医院检查确诊的PDR患者3例（PDR组）及非糖尿病患者3例（对照组）纳入研究。另外分别选取同期PDR、非糖尿病患者各40例并据此分为PDR验证组、对照验证组。PDR验证组、对照验证组行外周血验证试验；PDR组、对照组进行RNA测序。采用转录组学（RNAseq）测序技术筛选PDR组、对照组DEG。对筛选得到的DEG进行基因注释（GO）功能富集分析、京都基因与基因组百科全书（KEGG）的信号通路富集分析和蛋白-蛋白互作网络（PPI）分析。应用基因表达总集数据库查找与PDR相关高通量数据行多队列比对分析，通过Targetscan平台对差异表达的miRNA进行靶基因预测，明确筛选所得DEG与PDR的相关性。采用逆转录聚合酶链反应、蛋白免疫印迹法对与PDR相关的DEG mRNA、蛋白表达进行验证。PDR验证组、对照验证组之间PDR相关的DEG mRNA、蛋白相对表达量比较行配对n t检验。n 结果:RNAseq测序共筛选出1 337个DEG，上调基因419个，下调基因918个。具有低等电点的直接凋亡抑制蛋白结合蛋白（n DIABLO）、锌指和含BTB域10 （n ZBTB10 ）、polo样激酶3 （n PLK3 ）、调节亚单位1 （n PIK3R1）、B细胞易位基因3 （n BTG3）等基因在PDR组患者中差异表达。GO功能富集分析结果显示，n DIABLO、n ZBTB10、n PLK3、n PIK3R1、n BTG3等基因参与了与PDR相关的病理过程。KEGG富集分析结果显示，细胞外基质受体作用、细胞因子调控通路、p53信号通路、半乳糖代谢等糖代谢途径可能参与了DEG涉及过程。PPI分析结果提示，DEG关联蛋白节点越大，关联的节点数量越多，其中n DIABLO、n ZBTB10、n PLK3、n PIK3R1、n BTG3等基因对该作用网络形成作用显著。Targetscan平台预测发现，DR患者房水、玻璃体、血浆中显著差异miRNA与本研究所发现的DEG具有调控关系。与对照验证组比较，PDR验证组患者外周血中n DIABLO、n ZBTB10、n PLK3、n PIK3R1 mRNA、蛋白相对表达量上调，n BTG3 mRNA、蛋白相对表达量下调。n 结论:PDR患者的DEG为n DIABLO、n ZBTB10、n PLK3、n PIK3R1、n BTG3，其通过调控细胞增生、纤维化及氧化应激等生物学过程参与疾病进程。n “,”Objective:To screening differentially expressed genes (DEGs) in proliferative diabetic retinopathy (DR) patients to provide new biological therapeutic targets for proliferative DR (PDR) therapy.Methods:A basic research. A total of 3 PDR patients (group PDR) and 3 non-diabetic patients (control group) were enrolled in the study in Tianjin Medical University Eye Hospital in October 2020. In addition, 40 cases of PDR and non-diabetic patients were selected and divided into PDR validation group and control validation group. Peripheral blood validation test was performed in PDR validation group and control validation group; RNA sequencing was performed in PDR group and control group. Transcriptomics (RNAseq) sequencing technology was used to screen DEG in PDR group and control group. The selected DEGs were analyzed by gene ontology (GO) function enrichment analysis, signal pathway enrichment analysis of Kyoto Encyclopedia of Genes and Genomes (KEGG) and protein-protein interaction network (PPI). The gene expression database was used to find the high-throughput data related to PDR, and multi queue comparison analysis was carried out. The target genes of differentially expressed miRNAs were predicted through targetscan platform, so as to clearly screen the correlation between DEG and PDR. Reverse transcription polymerase chain reaction and Western blot were used to verify the expression of DEG mRNA and protein related to PDR. The relative expression of PDR related DEG mRNA and protein between PDR validation group and control validation group were compared by paired n t-test.n Results:A total of 1 337 DEGs were screened by RNAseq sequencing in the peripheral blood of patients with PDR, of which 419 genes were up-regulated and 918 down-regulated. Among them, direct inhibitor of apoptosis protein-binding protein with low isoelectric point (n DIABLO), zinc finger and BTB domain containing 10 (n ZBTB10), polo-like kinases 3 (n PLK3), regulatory subunit 1 (n PIK3R1) and B cell translocation gene 3 (BTG3) were differentially expressed in PDR patients. The function of GO was enriched from the analysis of molecular function, biological process and cellular composition. The results showed that n DIABLO, n ZBTB10, n PLK3, n PIK3R1, n BTG3 were involved in the pathological process related to PDR. KEGG enrichment analysis showed that glucose metabolic pathways such as extracellular matrix receptors, cytokine regulatory pathway, p53 signal pathway and galactose metabolism may be involved in the process of differential genes. The analysis of PPI protein interaction network showed that the larger the DEG-associated protein node, the greater the number of associated nodes. Among them, n DIABLO, n ZBTB10, n PLK3, n PIK3R1 and n BTG3 played significant roles in the formation of the action network. By comparing and analyzing the existing high-throughput data related to diabetic retinopathy in Gene Expression Omnibus database and predicting by Targetscan platform, it was found that some significant differences in miRNA reported in aqueous humor, vitreous fluid and plasma of DR patients can be regulated by the differential genes found in this study. Compared with the control verification group, the relative expressions of n DIABLO, ZBTB10, PLK3, PIK3R1 mRNA and protein in peripheral blood of the PDR verification group were up-regulated, and the relative expression of n BTG3 mRNA and protein was down-regulated.n Conclusion:DIABLO, n ZBTB10, n PLK3, n PIK3R1 and n BTG3 are DEGs in patients with PDR, and they can participate in the disease process by regulating the biological processes of cell proliferation, fibrosis and oxidative stress.n