Proliferative Diabetic Retinopathy (PDR) is a chronic complication of Diabetes and the main cause of blindness among the world’s working population at present. While there have been many studies on the pathogenesis of PDR, its intrinsic molecular mechanisms have not yet been fully elucidated. In recent years, several studies have employed bulk RNA-sequencing (RNA-seq) and single-cell RNA sequencing (scRNA-seq) to profile differentially expressed genes (DEGs) and cellular components associated with PDR. This study adds to this expanding body of work by identifying PDR’s target genes and cellular components by conducting an integrated transcriptome bioinformatics analysis. This study integrately examined two public bulk RNA-seq datasets(including 11 PDR patients and 7 controls) and one single-cell RNA-seq datasets(including 5 PDR patients) of Fibro (Vascular) Membranes (FVMs) from PDR patients and control. A total of 176 genes were identified as DEGs between PDR patients and control among both bulk RNA-seq datasets. Based on these DEGs, 14 proteins were identified in the protein overlap within the significant ligand-receptor interactions of retinal FVMs and Protein-Protein Interaction (PPI) network, three of which were associated with PDR (CD44, ICAM1, POSTN), and POSTN might act as key ligand. This finding may provide novel gene signatures and therapeutic targets for PDR.

Insulin therapy is still the most important treatment for T2DM, but the discussion about whether insulin brings more benefits or harms to T2DM patients has not stopped. Therefore, we used high-throughput RNA sequencing to investigate the role of insulin in T2DM and its molecular changes. We collected peripheral blood samples from 16 patients with T2DM, and performed RNA-seq on peripheral blood mononuclear cells. Bioinformatics analysis and machine learning were uesd to identify the key differential genes and transcription factor networks. In addition, we performed the flow cytometry and staining to observe ROS level and endothelial-monocyte adhesion in PBMCs of both groups. A total of 529 differential genes were identified by bioinformatics analysis. 8 genes were identified as key genes, among which IL-6 had high importance in the random forest model. In transcription factor analysis, IL-6, RETN, CTSG and ELANE have abundant transcriptional regulatory relationships. Flow cytometry showed that ROS production, phagocytosis, leukocyte adhesion in insulin treatment group were lower than that in non-insulin treatment group. Insulin therapy is bidirectional, it can cause islet B cell damage and vascular complications, but also can reduce the level of inflammation and oxidative stress.

Background: Diabetes increases the risk of cataract formation fivefold. Immune-mediated inflammation has been reported to play a role in this process; however, whether alterations in the immune landscape are involved remains unknown. Therefore, we conducted a multi-omics analysis to evaluate the impact of immune inflammation on the lens. Methods: Bulk RNA sequencing was performed on peripheral blood mononuclear cells (PBMCs) from diabetic patients and lens tissues from diabetic rats. Single-cell RNA sequencing was utilized to characterize intercellular interactions. Key gene and protein expressions were validated via laboratory assays. Results: An integrated RNA-seq analysis revealed a disruption of the blood–aqueous barrier integrity in the diabetic group, enhanced monocyte migration and adhesion, increased differentiation from classical to non-classical monocytes, and the upregulation of TNF and IFN-γ signaling pathways. The transcriptomic profiling of rat lenses revealed an increased proportion of monocytes and the activation of apoptotic pathways in lens epithelial cells. Immunohistochemistry and immunofluorescence staining demonstrated elevated caspase-3 and IL-6 levels in lens epithelial cells and increased immune cell infiltration in the diabetic group. The qRT-PCR and ELISA confirmed elevated levels of the pro-inflammatory cytokines IL-6 and IFN-γ, alongside reduced anti-inflammatory cytokine IL-10 in the peripheral blood and aqueous humor of diabetic patients. Conclusions: Diabetes alters the peripheral immune microenvironment and disrupts the blood–aqueous barrier, promoting intraocular inflammation and lens epithelial cell apoptosis, thereby accelerating cataract development.