Intravenous Treatment with a Long-Chain Omega-3 Lipid Emulsion Provides Neuroprotection in a Murine Model of Ischemic Stroke - A Pilot Study - PubMed (original) (raw)

Intravenous Treatment with a Long-Chain Omega-3 Lipid Emulsion Provides Neuroprotection in a Murine Model of Ischemic Stroke - A Pilot Study

Dirk Berressem et al. PLoS One. 2016.

Abstract

Single long-chain omega-3 fatty acids (e.g. docosahexaenoic acid (DHA) or eicosapentaenoic acid (EPA)) are known for their neuroprotective properties associated with ischemic stroke. This pilot study aimed to test the effectiveness of an acute treatment with a long-chain omega-3 lipid emulsion (Omegaven 10%®, OGV) that contains fish oil (DHA 18 mg/ml; EPA 21 mg/ml) and α-tocopherol (0.2 mg/ml) in a transient middle cerebral artery occlusion (MCAO) model of ischemic stroke in mice. For this purpose, female CD-1 mice were anesthetized and subjected to 90 minutes of MCAO. To reflect a clinically relevant situation for an acute treatment, either after induction of stroke or after reperfusion, a single dose of OGV was injected intravenously into the tail vein (5 ml/kg b.w.). A neurological severity score was used to assess motor function and neurological outcome. Stroke-related parameters were determined 24 hours after MCAO. Microdialysis was used to collect samples from extracellular space of the striatum. Mitochondrial function was determined in isolated mitochondria or dissociated brain cells. Inflammation markers were measured in brain homogenate. According to control experiments, neuroprotective effects could be attributed to the long-chain omega-3 content of the emulsion. Intravenous injection of OGV reduced size and severity of stroke, restored mitochondrial function, and prevented excitotoxic glutamate release. Increases of pro-inflammatory markers (COX-2 and IL-6) were attenuated. Neurological severity scoring and neurochemical data demonstrated that acute OGV treatment shortly after induction of stroke was most efficient and able to improve short-term neurological outcome, reflecting the importance of an acute treatment to improve the outcome. Summarising, acute treatment of stroke with a single intravenous dose of OGV provided strong neuroprotective effects and was most effective when given immediately after onset of ischemia. As OGV is an approved fishoil emulsion for parenteral nutrition in humans, our results may provide first translational data for a possible early management of ischemic stroke with administration of OGV to prevent further brain damage.

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Conflict of interest statement

This study was funded by Fresenius Kabi. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

Figures

Fig 1

Fig 1. Effects of OGV and control emulsions when applied at reperfusion.

Saline, Lipofundin® (LPF) and d-α-tocopheryl polyethylene glycol 1000 succinate (TPGS) in doses equal to OGV when injected at reperfusion (a.r.). (A) Infarct areas and (B) differences in grayscale for LPF, TPGS, and Omegaven 10% (OGV) vs. Saline; n = 6. Mean ± SEM, p*<0.05; p**<0.01; p***<0.001; One-Way ANOVA with Tukey post-test. (C) Representative striatal brain slices for determination of differences in grayscale for each group. Density of grayscale in a representative area from core of infarction (solid circle) was subtracted from corresponding grayscale of contralateral control area (dashed circle).

Fig 2

Fig 2. Marker of mitochondrial function after treatment at reperfusion.

Sham-operated mice. (Control) versus stroke control group that received saline (Stroke) and stroke treatment group that received OGV at reperfusion (a.r.). (A) Mitochondrial membrane potential (MMP)- and (B) Adenosine triphosphate (ATP)-levels as measured 24 hours after reperfusion in dissociated brain cells, n = 8; (C) Respiration [pmol oxygen/(s* mg protein)] of different complexes of the respiratory chain were determined in isolated mitochondria, n = 6; (D) Respiratory control ratio (RCR) that indicates the coupling of mitochondrial respiration chain and citrate synthase activity that represents a quantitative marker for mitochondrial mass, n = 6. Mean ± SEM, p*<0.05; p**<0.01; p***<0.001; One-Way ANOVA with Tukey post-test.

Fig 3

Fig 3. Effects of OGV when given at stroke (a.s.).

(A) Neurobehavioral assessment (refer to S1 Table) for sham-operated mice (Control) versus control group that received saline (Stroke) and treatment group that received OGV at stroke (a.s.), (B) Representative brain slices for determination of infarct volume and differences in grayscale. (C) Effect on infarct volume and grayscale levels, n = 8. Mean ± SEM, p*<0.05; p**<0.01; p***<0.001; (A) One-Way ANOVA with Tukey post-test, (C) t-test.

Fig 4

Fig 4. Marker of mitochondrial function after treatment at stroke.

Sham mice (Control) versus stroke control group that received saline (Stroke) and stroke treatment group that received OGV at stroke (a.s.). (A) Mitochondrial membrane potential (MMP)- and (B) Adenosine triphosphate (ATP)-levels as measured in dissociated brain cells 24 hours after reperfusion. Energy metabolite levels in striatal core region of stroke for (C) glucose and (D) glutamate as determined by microdialysis 30 minutes before (PRE), 90 minutes during (Stroke) and 30 minutes after (POST) stroke surgery, n = 8. Mean ± SEM, p*<0.05; p**<0.01;p***<0.001; One-Way ANOVA with Tukey post-test.

Fig 5

Fig 5. Western Blot analysis of brain homogenates.

Sham-operated mice (Control) versus control group that received saline (Stroke) and treatment group that received OGV at stroke (a.s.). (A) COX-2-, (B) IL-6-, and (C) IL-10 protein levels, n = 8. Mean ± SEM, p*<0.05; p**<0.01; p***<0.001; One-Way ANOVA with Tukey post-test.

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This study was partly supported by Fresenius Kabi Deutschland GmbH, Bad Homburg, Deutschland. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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