Docosahexaenoic acid alleviates cell injury and improves barrier function by suppressing necroptosis signalling in TNF-α-challenged porcine intestinal epithelial cells

Abstract

Long-chain n-3 polyunsaturated fatty acids are known to have beneficial effects on intestinal health. However, the underling mechanisms are largely unknown. The present study was conducted to investigate whether docosahexaenoic acid (DHA) attenuates TNF-α-induced intestinal cell injury and barrier dysfunction by modulating necroptosis signalling. Intestinal porcine epithelial cell line 1 was cultured with or without 12.5 µg/ml DHA, followed by exposure to 50 ng/ml TNF-α for indicated time periods. DHA restored cell viability and cell number triggered by TNF-α. DHA also improved barrier function, which was indicated by increased trans-epithelial electrical resistance, decreased FD4 flux and increased membrane localisation of zonula occludins (ZO-1) and claudin-1. Moreover, DHA suppressed cell necrosis in TNF-α-challenged cells, as shown in the IncuCyte ZOOM™ live cell imaging system and transmission electron microscopy. In addition, DHA decreased protein expression of TNF receptor, receptor interacting protein kinase 1, RIP3 and phosphorylation of mixed lineage kinase-like protein, phosphoglycerate mutase family 5, dynamin-related protein 1 and high mobility group box-1 protein. Furthermore, DHA suppressed protein expression of caspase-3 and caspase-8. Collectively, these results indicate that DHA is capable of alleviating TNF-α-induced cell injury and barrier dysfunction by suppressing the necroptosis signalling pathway.

Macrocystis pyrifera Lipids Reduce Cytokine-Induced Pro-Inflammatory Signalling and Barrier Dysfunction in Human Keratinocyte Models

“…In 3D epidermal constructs stimulated simultaneously with these Th1 and Th2 cytokines, treatment with the lipid extract partially restored claudin-1 protein density and tight junction localisation whilst reducing transepidermal leakage of lucifer yellow. These findings are in contrast with a previous report, where omega-3 fatty acids increased claudin-1 expression and localisation in intestinal cells treated with 2,4,6-trinitrobenzenesulfonic acid or TNF-α [68,69]. Together, these findings indicate that seaweed lipids may primarily exert barrier protective effects in the context of Th1-mediated inflammation.…”

“…leakage of lucifer yellow. These findings are in contrast with a previous report, wher omega-3 fatty acids increased claudin-1 expression and localisation in intestinal cell treated with 2,4,6-trinitrobenzenesulfonic acid or TNF-α [68,69]. Together, these finding indicate that seaweed lipids may primarily exert barrier protective effects in the contex of Th1-mediated inflammation.…”

(-)-Epigallocatechin-3-gallate promotes intestinal epithelial proliferation and barrier function after ischemia/reperfusion injury via activation of Nurr1

“…2020 ; Xiao et al. 2020 ). EGCG is the most abundant component and have strong physiological activities in green tea which has been considered to have beneficial properties for its anti-carcinogenetic, antioxidant, anti-inflammatory and anti-apoptotic effects (Khan et al 2008 ; Wu et al.…”

Effects of TolC on the pathogenicity of porcine extraintestinal pathogenic Escherichia coli

“…PAVECs were seeded in six-well plates and incubated with WT, ΔTolC, or Cm-TolC for 4 h, followed by treatment with PBS. Cells were then lysed and subjected to Western blotting as previously described ( 31 ). Blots were incubated with primary antibodies against rabbit anti-receptor-interacting protein kinase 1 (RIP1) (#LS-B8214, LifeSpan), rabbit anti-total mixed-lineage kinase domain-like protein (t-MLKL) (#37705, Cell Signaling Technology), rabbit anti-phosphoglycerate mutase family member 5 (PGAM5) (#ab131552, Abcam), and mouse anti-β-actin (#A2228, Sigma Aldrich).…”