Epac1 inhibits PKR to reduce NLRP3 inflammasome proteins in retinal endothelial cells

• Main Authors: Jiang Y, Steinle JJ
• Format: Article
• Language: English
• Published: Dove Medical Press 2019-06-01
• Series: Journal of Inflammation Research
• Subjects: inflammasome; NLRP3; PKR; retinal endothelial cells
• Online Access: https://www.dovepress.com/epac1-inhibits-pkr-to-reduce-nlrp3-inflammasome-proteins-in-retinal-en-peer-reviewed-article-JIR

Youde Jiang, Jena J SteinleDepartment of Ophthalmology, Visual and Anatomical Sciences, Wayne State University School of Medicine, Detroit, MI, USAPurpose: Inflammation has been strongly associated with retinal damage in diseases such as diabetic retinopathy. Several studies have reported that high glucose exposure induces damage to the retinal vasculature. We and others have shown that high glucose can activate the NOD-like receptor family, pyrin domain containing family member 3 (NLRP3) pathway, leading to increased levels of cleaved caspase 1 and IL-1β to activate a number of inflammatory pathways in the retina.Methods: We used retinal endothelial cells grown in normal (5 mM) or high (25 mM) glucose or retinal lysates from endothelial cell-specific knockout mice for exchange protein activated by cAMP 1 (Epac1). Human recombinant protein kinase R (PKR) or C16, a PKR inhibitor, was used on the cells to dissect PKR and NLRP3 signaling.Results: Using retinal endothelial cells (REC) in high glucose and whole retinal lysates from endothelial cell-specific knockout of Epac1, we demonstrate that Epac1 regulates PKR phosphorylation. Using an Epac1 agonist or PKR inhibition with C16, we demonstrated that loss of PKR resulted in reduced NLRP3, cleaved caspase 1, and IL-1β levels. Furthermore, despite the addition of recombinant human PKR, Epac1 was still able to significantly reduce NLRP3 signaling.Conclusion: Overall, these studies demonstrated that PKR regulates the NLRP3 inflammasome in REC, and that Epac1 inhibition of PKR can reduce activation of the NLRP3 inflammasome.Keywords: inflammasome, NLRP3, PKR, retinal endothelial cells