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Anti-inflammatory effects of epoxyeicosatrienoic acids (EETs) and their corresponding diols against pro-inflammatory cytokine-toxicity in BRIN-BD11 cells.

Watson

Authors

Watson



Abstract

Introduction: CYP450-derived EETs display anti-inflammatory activity in cardiac models of inflammation, in part through activation of Peroxisome proliferator-activated receptor gamma (PPARγ) and inhibition of Nuclear Factor kappa B (NF-kB) activation. Therefore, we aimed to investigate the cytoprotective effects of EET regioisomers and their corresponding vicinal diols in a beta cell model of pro-inflammatory cytokine toxicity.

Methods: BRIN-BD11 cells were treated with 100U/ml interleukin-1β, 20U/ml Interferon gamma (IFNγ) and 500U/ml tumour necrosis factor-α in co-incubation with either 10μM 8 (9)-EET, 11 (12)-EET, 14 (15)-EET, or their dihydroxyeicosatrienoic acid derivatives (DHETs) for 24h. Cell viability was assessed by vital dye exclusion (Trypan Blue; expressed as viable cells/ml) or multicaspase-activity assay and NF-kB activity was measured using a NanoLuc® luciferase reporter assay.

Results: All EETs protected against cytokine-induced cell death, such that cytokines decreased viable cell number from 0.79 × 106 to 0.33 × 106 and in co-incubation with 8 (9)-EET, 11 (12)-EET and 14 (15)-EET, this increased to 0.64 × 106, 0.61 × 106 and 0.59 × 106, respectively (p < 0.05). Similarly, cytokine treatment increased caspase activity to 35% (±5.7), decreasing to 18% (±1.9), 19% (±2) and 19% (±1.9) in the presence of these EETs (p < 0.05), accompanied by a 32% decrease in NF-kB activation. Of the corresponding diols only, 8 (9)-DHET attenuated cytokine toxicity, reducing caspase activity from 41% (±4.5) to 21% (±2.4) (p < 0.05).

Conclusion: EETs protected against cytokine toxicity in BRIN-BD11 cells, in part, via reduced activation of NF-kB. We also consider the novel observation that 8 (9)-DHET, unlike other EET-derived DHETs, similarly protected against cytokine-induced apoptosis. These data highlight a potential role of EETs in attenuating cytokine toxicity in Type 1 diabetes, and our ongoing work is exploring the production of endogenous EETs by CYP450 isoforms and actions of EET analogues.

Presentation Conference Type Poster
Conference Name Diabetes UK Professional Conference 2018
Conference Location London ExCeL, London
Start Date Mar 14, 2018
End Date Mar 16, 2018
Publication Date Dec 29, 2017
Publicly Available Date Mar 29, 2024
Publisher Wiley
Series Title Diabetes UK Annual Professional Conference
Publisher URL https://onlinelibrary.wiley.com/doi/10.1111/dme.3_13571