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

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.

Citation

Watson. (2018, March). Anti-inflammatory effects of epoxyeicosatrienoic acids (EETs) and their corresponding diols against pro-inflammatory cytokine-toxicity in BRIN-BD11 cells. Poster presented at Diabetes UK Professional Conference 2018, London ExCeL, London

• 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: May 26, 2023
• Publisher: Wiley
• Series Title: Diabetes UK Annual Professional Conference
• Publisher URL: https://onlinelibrary.wiley.com/doi/10.1111/dme.3_13571