Influence of endotoxin-mediated retinal inflammation on phenotype of diabetic retinopathy in Ins2Akita mice

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Abstract

Aims

To evaluate the impact of systemic exposure to bacterial lipopolysaccharide (LPS) on a rodent model of background diabetic retinopathy.

Methods

Toll-like receptor 4 (TLR4)-mediated systemic inflammation was induced in Ins2Akita heterozygotes and age-matched C57BL6/J-Ins2+ littermates by single or repeated intraperitoneal injections of the TLR4 ligand LPS (9 µg/g body weight). 24 hours after a single injection in 7-week-old mice retinal Il1b, Tnfa and Vegf transcripts were measured with real-time PCR. Vascular endothelial growth factor (VEGF) protein levels were evaluated with bead-based immunoassay. Leukostasis and endothelial injury were assessed in retinal wholemounts following perfusion with rhodamine or FITC conjugated concanavalin A to label leukocytes and propidium iodide to label dead or injured cells. In mice which had received three fortnightly injections between 10 and 16 weeks of age, retinal thicknesses and vascular structure were evaluated at 17–18 weeks of age using optical coherence tomography (OCT) and fluorescein angiography. Retinal architecture was assesed using resin-based histology.

Results

Compared with normoglycaemic controls, systemic LPS exposure in Ins2Akita mice was associated with a 3.5-fold increase in endothelial cell injury and attenuated leukostasis in the retinal vasculature. Hyperglycaemia or acute LPS inflammation did not increase retinal VEGF content. Thinning (10–13 µm) of posterior retina was detected with OCT 2 weeks after repeated exposure to LPS in Ins2Akita mice but not in normoglycaemic controls. Capillary networks and retinal morphology were unaffected by recurrent LPS inflammation in Ins2Akita and control mice.

Conclusions

In hyperglycaemic mice, exposure to systemic LPS was associated with two hallmark pathologies of early background diabetic retinopathy, namely, the injury of capillary endothelium and in vivo thinning of the retina.

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