The NLRP3 Inflammasome May Contribute to Pathologic Neovascularization in the Advanced Stages of Diabetic Retinopathy

• Published in: Scientific reports Vol. 8; no. 1; pp. 2847 - 15
• Main Authors: Chaurasia, Shyam S., Lim, Rayne R., Parikh, Bhav H., Wey, Yeo Sia, Tun, Bo Bo, Wong, Tien Yin, Luu, Chi D., Agrawal, Rupesh, Ghosh, Arkasubhra, Mortellaro, Alessandra, Rackoczy, Elizabeth, Mohan, Rajiv R., Barathi, Veluchamy A.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 12.02.2018; Nature Publishing Group; Nature Portfolio
• Subjects: 13/1; 13/21; 13/51; 14/1; 14/34; 38/39; 38/77; 631/378/2596/1953; 631/80/304; 64/60; 82/1; Angiogenesis; Angiography; Animals; Caspase; Caspase-1; CD14 antigen; CD31 antigen; Cell activation; Cytokines - genetics; Cytokines - metabolism; Diabetes; Diabetes mellitus; Diabetic retinopathy; Diabetic Retinopathy - diagnostic imaging; Diabetic Retinopathy - genetics; Diabetic Retinopathy - metabolism; Diabetic Retinopathy - pathology; Disease Models, Animal; Electroretinography; Enzyme-linked immunosorbent assay; Fluorescein; Fluorescein Angiography; Glia; Glial fibrillary acidic protein; Glial Fibrillary Acidic Protein - genetics; Glial Fibrillary Acidic Protein - metabolism; Humanities and Social Sciences; Humans; Hyperglycemia; Immunohistochemistry; Inflammasomes; Insulin - genetics; Intercellular adhesion molecule 1; Interleukin 6; Macrophages; Mice; Mice, Transgenic; Microglia; mRNA; multidisciplinary; Neovascularization, Pathologic - diagnostic imaging; Neovascularization, Pathologic - genetics; Neovascularization, Pathologic - metabolism; Neovascularization, Pathologic - pathology; NLR Family, Pyrin Domain-Containing 3 Protein - genetics; NLR Family, Pyrin Domain-Containing 3 Protein - metabolism; Photography; Retina; Retina - diagnostic imaging; Retina - pathology; Rodents; Science; Science (multidisciplinary); Tomography, Optical Coherence; Transgenic mice; Vascular endothelial growth factor; Vascular Endothelial Growth Factor A - genetics; Vascular endothelial growth factor receptor 2; Western blotting
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-018-21198-z

Diabetic retinopathy (DR) is a retinal microvascular disease characterized by inflammatory and angiogenic pathways. In this study, we evaluated NLRP3 inflammasome in a double transgenic mouse model, Akimba ( Ins2 Akita xVEGF + / − ), which demonstrates hyperglycemia, vascular hyperpermeability and neovascularization seen in the proliferative DR. Retinal structural integrity, vascular leakage and function were examined by fundus photography, fluorescein angiography, optical coherence tomography, retinal flat mounts, laser speckle flowgraphy (LSFG), and electroretinography in Akimba and its parental strains, Akita ( Ins2 Akita ) and Kimba ( trVEGF029 ) mice. Inflammatory mechanisms involving NLRP3 inflammasome were investigated using real time-PCR, immunohistochemistry, ELISA and western blots. We observed an increased vascular leakage, reduced retinal thickness, and function in Akimba retina. Also, Akimba retina depicts decreased relative flow volume measured by LSFG. Most importantly, high levels of IL-1β along with increased NLRP3, ASC, and Caspase-1 at mRNA and protein levels were observed in Akimba retina. However, the in vivo functional role remains undefined. In conclusion, increased activation of macroglia (GFAP), microglia (Iba-1 and OX-42) and perivascular macrophages (F4/80 and CD14) together with pro-inflammatory (IL-1β and IL-6) and pro-angiogenic markers (PECAM-1, ICAM-1, VEGF, Flt-1, and Flk-1), suggested a critical role for NLRP3 inflammasome in the Akimba mouse model depicting advanced stages of DR pathogenesis.