Journal Article

Thioredoxin-interacting protein is required for endothelial NLRP3 inflammasome activation and cell death in a rat model of high-fat diet

Ergul, Adviye, Mohamed, Islam N., Hafez, Sherif S., Imig, John D., Fairaq, Arwa, El-Remessy, Azza B.

2014

Overview

Aims/hypothesis Obesity and hypertension, known pro-inflammatory states, are identified determinants for increased retinal microvascular abnormalities. However, the molecular link between inflammation and microvascular degeneration remains elusive. Thioredoxin-interacting protein (TXNIP) is recognised as an activator of the NOD-like receptor pyrin domain containing-3 (NLRP3) inflammasome. This study aims to examine TXNIP expression and elucidate its role in endothelial inflammasome activation and retinal lesions. Methods Spontaneously hypertensive (SHR) and control Wistar (W) rats were compared with groups fed a high-fat diet (HFD) (W+F and SHR+F) for 8–10 weeks. Results Compared with W controls, HFD alone or in combination with hypertension significantly induced formation of acellular capillaries, a hallmark of retinal ischaemic lesions. These effects were accompanied by significant increases in lipid peroxidation, nitrotyrosine and expression of TXNIP, nuclear factor κB, TNF-α and IL-1β. HFD significantly increased interaction of TXNIP–NLRP3 and expression of cleaved caspase-1 and cleaved IL-1β. Immunolocalisation studies identified TXNIP expression within astrocytes and Müller cells surrounding retinal endothelial cells. To model HFD in vitro, human retinal endothelial (HRE) cells were stimulated with 400 μmol/l palmitate coupled to BSA (Pal-BSA). Pal-BSA triggered expression of TXNIP and its interaction with NLRP3, resulting in activation of caspase-1 and IL-1β in HRE cells. Silencing Txnip expression in HRE cells abolished Pal-BSA-mediated cleaved IL-1β release into medium and cell death, evident by decreases in cleaved caspase-3 expression and the proportion of live to dead cells. Conclusions/interpretation These findings provide the first evidence for enhanced TXNIP expression in hypertension and HFD-induced retinal oxidative/inflammatory response and suggest that TXNIP is required for HFD-mediated activation of the NLRP3 inflammasome and the release of IL-1β in endothelial cells.

• Publisher: Springer Berlin Heidelberg,Springer,Springer Nature B.V
• Subject: Animals / Arterial hypertension. Arterial hypotension / Biological and medical sciences / Blood and lymphatic vessels / Blood vessels / Cardiology. Vascular system / Carrier Proteins - biosynthesis / Carrier Proteins - metabolism / Caspase Inhibitors - metabolism / Cell Cycle Proteins / Cell Death / Cytokines / Diabetes / Diabetes. Impaired glucose tolerance / Diet, High-Fat / Endocrine pancreas. Apud cells (diseases) / Endocrinopathies / Endothelial Cells - metabolism / Etiopathogenesis. Screening. Investigations. Target tissue resistance / Experimental diseases / Eye and associated structures. Visual pathways and centers. Vision / Eye Diseases - metabolism / Eye Diseases - pathology / Fundamental and applied biological sciences. Psychology / Human Physiology / Hypertension / Hypotheses / Inflammasomes - metabolism / Inflammation / Inflammation - metabolism / Insulin resistance / Internal Medicine / Male / Medical sciences / Medicine / Medicine & Public Health / Metabolic Diseases / Metabolic syndrome / Microcirculation / NF-kappa B - metabolism / NLR Family, Pyrin Domain-Containing 3 Protein / Obesity / Obesity - complications / Obesity - metabolism / Oxidative Stress / Proteins / Rats / Rats, Wistar / Reagents / Receptors, Cytoplasmic and Nuclear - metabolism / Retina - metabolism / Retina - pathology / Vertebrates: nervous system and sense organs