Interferon Regulatory Factor 1 Promotes Diabetic Vasculopathy by Mediating Endothelial Cell Pyroptosis

Abstract

Background

Diabetic vasculopathy is not only one of the most common and serious complications of diabetes mellitus but also the leading cause of death in patients with type 2 diabetes mellitus (T2DM). Endothelial cells, located at the interface between blood and interstitial tissues, are the first line of defense of the vascular system against inflammatory damage, and their death is usually linked to atherosclerosis progression. However, the exact mechanisms need to be explored in greater depth.

Methods

GSE169332 data, including one group of no diabetic atherosclerosis and one group of diabetic atherosclerosis, were integrated and analyzed using single-cell RNA sequencing technology. The biological pathways involved and the key factor interferon regulatory factor 1 (IRF-1) were explored by Gene Ontology pathway enrichment and trajectory analysis. The role of IRF-1-mediated endothelial cell pyroptosis in diabetic model mice BKS^−db/db was verified by in vivo assays of immunofluorescence, immunohistochemistry, hematoxylin and eosin staining, and Masson staining.

Results

Single-cell RNA sequencing results showed that the development of diabetic vasculopathy was associated with endothelial cell pyroptosis, and endothelial pyroptosis 1 (EP1) cells with high expression of pyroptosis-associated factors exhibited a significant increase in diabetic atherosclerosis samples. The Gene Ontology enrichment results further emphasized the importance of endothelial cells in the progression of diabetic vasculopathy, especially EP1 cells. Trajectory analysis revealed that IRF-1 was more likely to be the hub gene of EP1 relative to other pyroptosis-related genes. Immunofluorescence, immunohistochemistry, hematoxylin and eosin staining, and Masson staining of mouse vascular tissues demonstrated that IRF-1 expression was significantly increased in diabetic model mice BKS^−db/db, and the expression of pyroptosis-related marker the NACHT, LRR, and PYD domains-containing protein 3 (NLRP3), as well as interleukin-1beta (IL-1β) and interleukin-18 (IL-18) levels, was also significantly increased.

Conclusions

IRF-1 promotes the progression of diabetic vasculopathy by mediating endothelial cell pyroptosis through increasing the expression of pyroptosis-related marker NLRP3 and the levels of IL-1β and IL-18.