Impdh1 was identified as a key protein promotes diabetic vasculopathy by intervention of vascular endothelial cell pyroptosis

Abstract

Abstract Background Diabetic angiopathy (DA) is a diabetic vascular complication. Pyroptosis is an inflammatory death that plays an important role in the development of DA, but the underlying mechanisms have not been fully elucidated. Methods The GSE169332 dataset from the Gene Expression Omnibus (GEO) was subjected to single-cell RNA sequencing (scRNA-seq) analysis, and the data of diabetic mice were subjected to bulk RNA-seq. The pathway through which the inflammatory microenvironment participated in the DA was explored by pseudotime analysis and cell-cell communication. DA models were constructed using in vitro mouse models. The histopathological changes in the collected aorta were observed by hematoxylin and eosin (H&E) and Masson staining. The distribution and expression of the phenotypic markers related to pyroptosis in aortic tissues (NLRP3, pro-Caspase1, and GSDMD-N) were observed by immunohistochemistry (IHC) or immunofluorescence (IF) staining. Following the silencing of the expression of high glucose (HG)-induced Impdh1 in endothelial cells (ECs), Impdh1 expression was detected by real-time quantitative reverse transcription PCR (qRT-PCR), and the expression of Impdh1, NLRP3, pro-Caspase1, and GSDMD was detected by IF staining; cell migration was detected by cell scratch assay, cell viability was detected by cell counting kit-8 (CCK-8) assay, and tube formation was detected by tube formation assay; the levels of IL-1β and IL-18 were detected using the enzyme-linked immunosorbent assay (ELISA) kits. Results Impdh1 was identified by scRNA-seq and bulk RNA-seq as a key molecule in the progression of DA associated with pyroptosis of aortic ECs. By constructing mouse models of DA, it was found that silencing Impdh1 can inhibit mouse aortic pyroptosis. Silencing of the expression of HG-induced Impdh1 revealed an effective amelioration of EC damage and pyroptosis. Conclusion Impdh1 is identified as a potential pyroptosis-related gene associated with DA by scRNA-seq of GEO data and bulk RNA-seq. Impdh1 protects aortic ECs by inhibiting pyroptosis and inflammation. Clinical trial number Not applicable.

Keywords

• Diabetic angiopathy
• Aortic endothelial cells
• Pyroptosis
• Single-cell RNA sequencing
• Impdh1