Journal Article

Retinal inflammation in murine models of type 1 and type 2 diabetes with diabetic retinopathy

Dharmarajan, Subramanian, Sheibani, Nader, Wilson, Jonathan M., Baucum, Anthony J., Carrillo, Casandra, Sorenson, Christine M., Belecky-Adams, Teri L., Qi, Zhonghua

2023

Overview

Aims/hypothesis The loss of pericytes surrounding the retinal vasculature in early diabetic retinopathy underlies changes to the neurovascular unit that lead to more destructive forms of the disease. However, it is unclear which changes lead to loss of retinal pericytes. This study investigated the hypothesis that chronic increases in one or more inflammatory factors mitigate the signalling pathways needed for pericyte survival. Methods Loss of pericytes and levels of inflammatory markers at the mRNA and protein levels were investigated in two genetic models of diabetes, Ins2 Akita/+ (a model of type 1 diabetes) and Lepr db/db (a model of type 2 diabetes), at early stages of diabetic retinopathy. In addition, changes that accompany gliosis and the retinal vasculature were determined. Finally, changes in retinal pericytes chronically incubated with vehicle or increasing amounts of IFNγ were investigated to determine the effects on pericyte survival. The numbers of pericytes, microglia, astrocytes and endothelial cells in retinal flatmounts were determined by immunofluorescence. Protein and mRNA levels of inflammatory factors were determined using multiplex ELISAs and quantitative reverse transcription PCR (qRT-PCR). The effects of IFNγ on the murine retinal pericyte survival-related platelet-derived growth factor receptor β (PDGFRβ) signalling pathway were investigated by western blot analysis. Finally, the levels of cell death-associated protein kinase C isoform delta (PKCδ) and cleaved caspase 3 (CC3) in pericytes were determined by western blot analysis and immunocytochemistry. Results The essential findings of this study were that both type 1 and 2 diabetes were accompanied by a similar progression of retinal pericyte loss, as well as gliosis. However, inflammatory factor expression was dissimilar in the two models of diabetes, with peak expression occurring at different ages for each model. Retinal vascular changes were more severe in the type 2 diabetes model. Chronic incubation of murine retinal pericytes with IFNγ decreased PDGFRβ signalling and increased the levels of active PKCδ and CC3. Conclusions/interpretation We conclude that retinal inflammation is involved in and sustains pericyte loss as diabetic retinopathy progresses. Moreover, IFNγ plays a critical role in reducing pericyte survival in the retina by reducing activation of the PDGFRβ signalling pathway and increasing PKCδ levels and pericyte apoptosis. Graphical Abstract

• Publisher: Springer Berlin Heidelberg,Springer Nature B.V
• Subject: Animal models / Animals / Apoptosis / Astrocytes / Caspase-3 / Cell death / Death-associated protein kinase / Diabetes / Diabetes mellitus (insulin dependent) / Diabetes mellitus (non-insulin dependent) / Diabetes Mellitus, Experimental - metabolism / Diabetes Mellitus, Type 1 - metabolism / Diabetes Mellitus, Type 2 - metabolism / Diabetic retinopathy / Diabetic Retinopathy - genetics / Diabetic Retinopathy - metabolism / Disease Models, Animal / Endothelial cells / Endothelial Cells - metabolism / Gliosis / Gliosis - complications / Gliosis - metabolism / Human Physiology / Hypotheses / Immunocytochemistry / Immunofluorescence / Inflammation / Inflammation - metabolism / Internal Medicine / Kinases / Medicine / Medicine & Public Health / Metabolic Diseases / Mice / Microglia / Pericytes / Pericytes - metabolism / Platelet-derived growth factor / Platelet-derived growth factor receptors / Protein kinase C / Proteins / Retina / Retina - metabolism / Retinopathy / Reverse transcription / RNA, Messenger - genetics / RNA, Messenger - metabolism / Signal transduction / γ-Interferon