关键词: 糖尿病视网膜病变, 转录组学, 基因

Abstract: Objective To study the differentially expressed genes and DR-related genes in patients with diabetic retinopathy (DR) by using transcriptome technology. Design Prospective comparative case series. Participants Fourteen patients with T2DM were recuited, among them, eight patients with moderate or higher non-proliferative DR (NPDR) or proliferative DR (PDR) were enrolled in the DR group and six patients without DR were enrolled in the DM group. Methods Peripheral blood of each patient was collected, white blood cells were isolated, total RNA was extracted, and retranscribed to DNA. Transcription group library was constructed. Illumina HiSeqTM2000 system was used to sequence the results. Bioinformatics analysis was used to analyze the sequencing results. RT-q PCR was used to detect the levels of calmodulin 1 (CNN1) and lysophosphatidic acid receptor 3 (LPAR3) in peripheral blood leukocytes. To validate transcriptome sequencing results. Main Outcome Measures Differentially expressed genes, GO annotation and enrichment, biochemical metabolism and pathway annotation and enrichment, and differential gene interaction network between the two groups. The levels of CNN1 and LPAR3 in peripheral leukocytes were different between the two groups. Results Compared with DM group, DR group screened 103 differentially expressed genes. Among them, 41 were up-regulated and 62 were down-regulated. The most significant down-regulation was killer cell immunoglobulin like receptor 2DS1 (KIR2DS1), and the most significant up-regulation was small nucleolar RNA gene (RNU105B). GO enrichment analysis showed that the GO items significantly enriched in biological process (GO-P) and molecular function (GO-F) were cell process, single biological process and binding function. Glutathione S-transferase mu1 (GSTM1), early growth response 1 (EGR-1), LPAR3 and CNN1 were all enriched in the above items. The analysis of significance enrichment of pathway showed that the most abundant differential genes were antigen treatment and presentation, natural killer cell-mediated cytotoxicity pathway, in which KIR2DS1 was involved. The analysis of gene interaction network showed that there was interaction between GAS1 and multiple genes. qPCR results indicated that the levels of CNN1 and LPAR3 in DR group were higher than those in DM group (all P＜0.05). Conclusion Using transcriptome technology, we found that there were many differential expression genes in peripheral blood leukocytes of DR group. Among them, whether KIR2DS1, GSTM1, EGR-1, and GAS1 in the down regulated genes, and RNU105B, LPAR3, and CNN1 in the up regulated genes are related to the occurrence and development of DR remains to be further verified. （Ophthalmol CHN， 2020, 29： 105-113）

Key words: diabetic retinopathy, transcriptome, gene