Protective effects of omega-3 fatty acids in a blood-brain barrier-on-chip model and on postoperative delirium-like behaviour in mice.

BACKGROUND: Peripheral surgical trauma can trigger neuroinflammation and ensuing neurological complications, such as delirium. The mechanisms whereby surgery contributes to postoperative neuroinflammation remain unclear and without effective therapies. Here, we developed a microfluidic-assisted blood-brain barrier (BBB) device and tested the effects of omega-3 fatty acids on neuroimmune interactions after orthopaedic surgery. METHODS: A microfluidic-assisted BBB device was established using primary human cells. Tight junction proteins, vascular cell adhesion molecule 1 (VCAM-1), BBB permeability, and astrocytic networks were assessed after stimulation with interleukin (IL)-1ß and in the presence or absence of a clinically available omega-3 fatty acid emulsion (Omegaven®; Fresenius Kabi, Bad Homburg, Germany). Mice were treated 1 h before orthopaedic surgery with 10 µl g-1 body weight of omega-3 fatty acid emulsion i.v. or equal volumes of saline. Changes in pericytes, perivascular macrophages, BBB opening, microglial activation, and inattention were evaluated. RESULTS: Omega-3 fatty acids protected barrier permeability, endothelial tight junctions, and VCAM-1 after exposure to IL-1ß in the BBB model. In vivo studies confirmed that omega-3 fatty acid treatment inhibited surgery-induced BBB impairment, microglial activation, and delirium-like behaviour. We identified a novel role for pericyte loss and perivascular macrophage activation in mice after surgery, which were rescued by prophylaxis with i.v. omega-3 fatty acids. CONCLUSIONS: We present a new approach to study neuroimmune interactions relevant to perioperative recovery using a microphysiological BBB platform. Changes in barrier function, including dysregulation of pericytes and perivascular macrophages, provide new targets to reduce postoperative delirium.

Agathisflavone isolated from Anacardium occidentale suppresses SIRT1-mediated neuroinflammation in BV2 microglia and neurotoxicity in APPSwe-transfected SH-SY5Y cells.

Agathisflavone is a bioactive compound in Anacardium occidentale. In this study, we investigated inhibition neuroinflammation in BV2 microglia by agathisflavone. Neuroprotective activity of the compound was investigated in differentiated SH-SY5Y cells. Experiments in lipopolysaccharide (LPS)-activated BV2 microglia showed that pretreatment with agathisflavone (5-20 µM) produced significant reduction in the release of tumour necrosis factor-α, interleukin-6, interleukin-1ß, NO, and PGE2 from the cells. Immunoblotting experiments also revealed that agathisflavone reduced levels of iNOS and COX-2 protein. Further studies revealed that agathisflavone reduced neuroinflammation by targeting critical steps in NF-κB signalling in BV2 microglia. Treatment of SH-SY5Y cells with conditioned medium from LPS-activated BV2 microglia produced a significant reduction in neuronal viability. However, conditioned medium from BV2 cells that were stimulated with LPS in the presence of agathisflavone did not induce neurotoxicity. Agathisflavone also produced neuroprotection in APPSwe plasmid-transfected SH-SY5Y neurons. The compound further attenuated LPS-induced and APPSwe plasmid-induced reduction in SIRT1 in BV2 microglia and SH-SY5Y, respectively. In the presence of EX527, agathisflavone lost its anti-inflammatory and neuroprotective activities. Our results suggest that agathisflavone inhibits neuroinflammation in BV2 microglia by targeting NF-κB signalling pathway. The compound also reduces neurotoxicity through mechanisms that are possibly linked to SIRT1 in the microglia and neurons.

Pomegranate inhibits neuroinflammation and amyloidogenesis in IL-1ß-stimulated SK-N-SH cells.

PURPOSE: Pomegranate fruit, Punica granatum L. (Punicaceae), and its constituents have been shown to inhibit inflammation. In this study, we aimed to assess the effects of freeze-dried pomegranate (PWE) on PGE2 production in IL-1ß-stimulated SK-N-SH cells. METHODS: An enzyme immunoassay (EIA) was used to measure prostaglandin E2 (PGE2) production from supernatants of IL-1ß-stimulated SK-N-SH cells. Expression of COX-2, phospho-IκB, and phospho-IKK proteins was evaluated, while NF-κB reporter gene assay was carried out in TNFα-stimulated HEK293 cells to determine the effect of PWE on NF-κB transactivation. Levels of BACE-1 and Aß in SK-N-SH cells stimulated with IL-1ß were measured with an in cell ELISA. RESULTS: PWE (25-200 µg/ml) dose dependently reduced COX-2-dependent PGE2 production in SK-N-SH cells stimulated with IL-1ß. Phosphorylation of IκB and IKK was significantly (p < 0.001) inhibited by PWE (50-200 µg/ml). Our studies also show that PWE (50-200 µg/ml) significantly (p < 0.01) inhibited NF-κB transactivation in TNFα-stimulated HEK293 cells. Furthermore, PWE inhibited BACE-1 and Aß expression in SK-N-SH cells treated with IL-1ß. CONCLUSIONS: Taken together, our study demonstrates that pomegranate inhibits inflammation, as well as amyloidogenesis in IL-1ß-stimulated SK-N-SH cells. We propose that pomegranate is a potential nutritional strategy in slowing the progression of neurodegenerative disorders such as Alzheimer's disease.

Picralima nitida seeds suppress PGE2 production by interfering with multiple signalling pathways in IL-1ß-stimulated SK-N-SH neuronal cells.

ETHNOPHARMACOLOGICAL RELEVANCE: The dried seed of Picralima nitida is used in rheumatic fever and as an antipyretic in West Africa. In this study we have investigated the effects of an extract obtained from the seeds of Picralima nitida (PNE) on PGE2 production in IL-1ß-stimulated cells. MATERIALS AND METHODS: Prostaglandin E2 (PGE2) was measured in supernatants of IL-1ß-stimulated SK-N-SH cells using enzyme immunoassay (EIA) for PGE2. In Cell ELISA and western blot were used to evaluate the effects of PNE on protein expressions of COX-2, mPGES-1, IκB and IKK. To determine the effect of the extract on NF-κB transactivation, a reporter gene assay was carried out in HEK293 cells stimulated with TNFα. An ELISA was used to measure the roles of p38, ERK1/2 and JNK Mitogen Activated Protein Kinases (MAPKs) on anti-neuroinflammatory actions of PNE. RESULTS: Results show that PNE significantly inhibited PGE2 production, as well as COX-2 and mPGES-1 protein expressions in IL-1ß-stimulated SK-N-SH cells. Molecular targeting experiments showed that PNE interfered with NF-κB signalling pathway through attenuation of TNFα-stimulated NF-κB transcriptional activation in HEK 293 cells. Furthermore, IL-1ß-mediated phosphorylation of IκB and IKK were inhibited in SK-N-SH cells. PNE (50-200 µg/ml) also produced significant inhibition of IL-1ß-induced p38 MAPK phosphorylation in SK-N-SH cells. However, phosphorylation of ERK1/2 and JNK MAPKs were achieved at 100 and 200 µg/ml of the extract. CONCLUSIONS: Taken together, these results clearly demonstrate that Picralima nitida suppresses PGE2 production by targeting multiple pathways involving NF-κB and MAPK signalling in IL-1ß-stimulated SK-N-SH neuronal cells.