Fumarate decreases edema volume and improves functional outcome after experimental stroke

Bettina Hjelm Clausen, Louise Lundberg, Minna Liisa Kyllikki Yli-Karjanmaa, Nellie Anne Martin, Martina Svensson, Maria Zeiler Alfsen, Simon Bertram Flæng, Kristina Sanne Lyngsø, Antonio Boza-Serrano, Helle Hvilsted Nielsen, Pernille B. Lærkegaard Hansen, Bente Finsen, Tomas Deierborg, Zsolt Illés, Kate Lykke Lambertsen

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Background Oxidative stress and inflammation exacerbate tissue damage in the brain after ischemic stroke. Dimethyl-fumarate (DMF) and its metabolite monomethyl-fumarate (MMF) are known to stimulate anti-oxidant pathways and modulate inflammatory responses. Considering these dual effects of fumarates, we examined the effect of MMF treatment after ischemic stroke in mice. Methods Permanent middle cerebral artery occlusion (pMCAO) was performed using adult, male C57BL/6 mice. Thirty minutes after pMCAO, 20 mg/kg MMF was administered intravenously. Outcomes were evaluated 6, 24 and 48 h after pMCAO. First, we examined whether a bolus of MMF was capable of changing expression of kelch-like erythroid cell-derived protein with CNC homology-associated protein 1 (Keap1) and nuclear factor erythroid 2-related factor (Nrf)2 in the infarcted brain. Next, we studied the effect of MMF on functional recovery. To explore mechanisms potentially influencing functional changes, we examined infarct volumes, edema formation, the expression of heat shock protein (Hsp)72, hydroxycarboxylic acid receptor 2 (Hcar2), and inducible nitric oxide synthase (iNOS) in the infarcted brain using real-time PCR and Western blotting. Concentrations of a panel of pro- and anti-inflammatory cytokines (IFNγ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12p70, TNF) were examined in both the infarcted brain tissue and plasma samples 6, 24 and 48 h after pMCAO using multiplex electrochemoluminiscence analysis. Results Administration of MMF increased the protein level of Nrf2 6 h after pMCAO, and improved functional outcome at 24 and 48 h after pMCAO. MMF treatment did not influence infarct size, however reduced edema volume at both 24 and 48 h after pMCAO. MMF treatment resulted in increased Hsp72 expression in the brain 6 h after pMCAO. Hcar2 mRNA levels increased significantly 24 h after pMCAO, but were not different between saline- and MMF-treated mice. MMF treatment also increased the level of the anti-inflammatory cytokine IL-10 in the brain and plasma 6 h after pMCAO, and additionally reduced the level of the pro-inflammatory cytokine IL-12p70 in the brain at 24 and 48 h after pMCAO. Conclusions A single intravenous bolus of MMF improved sensory-motor function after ischemic stroke, reduced edema formation, and increased the levels of the neuroprotective protein Hsp72 in the brain. The early increase in IL-10 and reduction in IL-12p70 in the brain combined with changes in systemic cytokine levels may also contribute to the functional recovery after pMCAO.

Original languageEnglish
JournalExperimental Neurology
Volume295
Pages (from-to)144–154
ISSN0014-4886
DOIs
Publication statusPublished - Sep 2017

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Middle Cerebral Artery Infarction
HSP72 Heat-Shock Proteins
Interleukin-10
Erythroid Cells
Proteins
Acids
Interleukin-5
Oxidants
Interleukin-4
Interleukin-2
Real-Time Polymerase Chain Reaction
Interleukin-6

Keywords

  • Cytokines
  • Dimethyl-fumarate
  • Focal cerebral ischemia
  • Heat shock protein
  • Monomethyl-fumarate
  • Nrf2
  • Tecfidera
  • Brain Ischemia/drug therapy
  • Kelch-Like ECH-Associated Protein 1/biosynthesis
  • Stroke/complications
  • Mice, Inbred C57BL
  • NF-E2-Related Factor 2/biosynthesis
  • Male
  • Treatment Outcome
  • Brain Edema/drug therapy
  • Heat-Shock Proteins/biosynthesis
  • Nitric Oxide Synthase Type II/antagonists & inhibitors
  • Dimethyl Fumarate/therapeutic use
  • Animals
  • Behavior, Animal/drug effects
  • Mice
  • Infarction, Middle Cerebral Artery/drug therapy
  • Cytokines/biosynthesis

Cite this

Clausen, Bettina Hjelm ; Lundberg, Louise ; Yli-Karjanmaa, Minna Liisa Kyllikki ; Martin, Nellie Anne ; Svensson, Martina ; Zeiler Alfsen, Maria ; Flæng, Simon Bertram ; Lyngsø, Kristina Sanne ; Boza-Serrano, Antonio ; Nielsen, Helle Hvilsted ; Hansen, Pernille B. Lærkegaard ; Finsen, Bente ; Deierborg, Tomas ; Illés, Zsolt ; Lambertsen, Kate Lykke. / Fumarate decreases edema volume and improves functional outcome after experimental stroke. In: Experimental Neurology. 2017 ; Vol. 295. pp. 144–154.
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title = "Fumarate decreases edema volume and improves functional outcome after experimental stroke",
abstract = "Background Oxidative stress and inflammation exacerbate tissue damage in the brain after ischemic stroke. Dimethyl-fumarate (DMF) and its metabolite monomethyl-fumarate (MMF) are known to stimulate anti-oxidant pathways and modulate inflammatory responses. Considering these dual effects of fumarates, we examined the effect of MMF treatment after ischemic stroke in mice. Methods Permanent middle cerebral artery occlusion (pMCAO) was performed using adult, male C57BL/6 mice. Thirty minutes after pMCAO, 20 mg/kg MMF was administered intravenously. Outcomes were evaluated 6, 24 and 48 h after pMCAO. First, we examined whether a bolus of MMF was capable of changing expression of kelch-like erythroid cell-derived protein with CNC homology-associated protein 1 (Keap1) and nuclear factor erythroid 2-related factor (Nrf)2 in the infarcted brain. Next, we studied the effect of MMF on functional recovery. To explore mechanisms potentially influencing functional changes, we examined infarct volumes, edema formation, the expression of heat shock protein (Hsp)72, hydroxycarboxylic acid receptor 2 (Hcar2), and inducible nitric oxide synthase (iNOS) in the infarcted brain using real-time PCR and Western blotting. Concentrations of a panel of pro- and anti-inflammatory cytokines (IFNγ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12p70, TNF) were examined in both the infarcted brain tissue and plasma samples 6, 24 and 48 h after pMCAO using multiplex electrochemoluminiscence analysis. Results Administration of MMF increased the protein level of Nrf2 6 h after pMCAO, and improved functional outcome at 24 and 48 h after pMCAO. MMF treatment did not influence infarct size, however reduced edema volume at both 24 and 48 h after pMCAO. MMF treatment resulted in increased Hsp72 expression in the brain 6 h after pMCAO. Hcar2 mRNA levels increased significantly 24 h after pMCAO, but were not different between saline- and MMF-treated mice. MMF treatment also increased the level of the anti-inflammatory cytokine IL-10 in the brain and plasma 6 h after pMCAO, and additionally reduced the level of the pro-inflammatory cytokine IL-12p70 in the brain at 24 and 48 h after pMCAO. Conclusions A single intravenous bolus of MMF improved sensory-motor function after ischemic stroke, reduced edema formation, and increased the levels of the neuroprotective protein Hsp72 in the brain. The early increase in IL-10 and reduction in IL-12p70 in the brain combined with changes in systemic cytokine levels may also contribute to the functional recovery after pMCAO.",
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author = "Clausen, {Bettina Hjelm} and Louise Lundberg and Yli-Karjanmaa, {Minna Liisa Kyllikki} and Martin, {Nellie Anne} and Martina Svensson and {Zeiler Alfsen}, Maria and Fl{\ae}ng, {Simon Bertram} and Lyngs{\o}, {Kristina Sanne} and Antonio Boza-Serrano and Nielsen, {Helle Hvilsted} and Hansen, {Pernille B. L{\ae}rkegaard} and Bente Finsen and Tomas Deierborg and Zsolt Ill{\'e}s and Lambertsen, {Kate Lykke}",
year = "2017",
month = "9",
doi = "10.1016/j.expneurol.2017.06.011",
language = "English",
volume = "295",
pages = "144–154",
journal = "Experimental Neurology",
issn = "0014-4886",
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Fumarate decreases edema volume and improves functional outcome after experimental stroke. / Clausen, Bettina Hjelm; Lundberg, Louise; Yli-Karjanmaa, Minna Liisa Kyllikki; Martin, Nellie Anne; Svensson, Martina; Zeiler Alfsen, Maria; Flæng, Simon Bertram; Lyngsø, Kristina Sanne; Boza-Serrano, Antonio; Nielsen, Helle Hvilsted; Hansen, Pernille B. Lærkegaard; Finsen, Bente; Deierborg, Tomas; Illés, Zsolt; Lambertsen, Kate Lykke.

In: Experimental Neurology, Vol. 295, 09.2017, p. 144–154.

Research output: Contribution to journalJournal articleResearchpeer-review

TY - JOUR

T1 - Fumarate decreases edema volume and improves functional outcome after experimental stroke

AU - Clausen, Bettina Hjelm

AU - Lundberg, Louise

AU - Yli-Karjanmaa, Minna Liisa Kyllikki

AU - Martin, Nellie Anne

AU - Svensson, Martina

AU - Zeiler Alfsen, Maria

AU - Flæng, Simon Bertram

AU - Lyngsø, Kristina Sanne

AU - Boza-Serrano, Antonio

AU - Nielsen, Helle Hvilsted

AU - Hansen, Pernille B. Lærkegaard

AU - Finsen, Bente

AU - Deierborg, Tomas

AU - Illés, Zsolt

AU - Lambertsen, Kate Lykke

PY - 2017/9

Y1 - 2017/9

N2 - Background Oxidative stress and inflammation exacerbate tissue damage in the brain after ischemic stroke. Dimethyl-fumarate (DMF) and its metabolite monomethyl-fumarate (MMF) are known to stimulate anti-oxidant pathways and modulate inflammatory responses. Considering these dual effects of fumarates, we examined the effect of MMF treatment after ischemic stroke in mice. Methods Permanent middle cerebral artery occlusion (pMCAO) was performed using adult, male C57BL/6 mice. Thirty minutes after pMCAO, 20 mg/kg MMF was administered intravenously. Outcomes were evaluated 6, 24 and 48 h after pMCAO. First, we examined whether a bolus of MMF was capable of changing expression of kelch-like erythroid cell-derived protein with CNC homology-associated protein 1 (Keap1) and nuclear factor erythroid 2-related factor (Nrf)2 in the infarcted brain. Next, we studied the effect of MMF on functional recovery. To explore mechanisms potentially influencing functional changes, we examined infarct volumes, edema formation, the expression of heat shock protein (Hsp)72, hydroxycarboxylic acid receptor 2 (Hcar2), and inducible nitric oxide synthase (iNOS) in the infarcted brain using real-time PCR and Western blotting. Concentrations of a panel of pro- and anti-inflammatory cytokines (IFNγ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12p70, TNF) were examined in both the infarcted brain tissue and plasma samples 6, 24 and 48 h after pMCAO using multiplex electrochemoluminiscence analysis. Results Administration of MMF increased the protein level of Nrf2 6 h after pMCAO, and improved functional outcome at 24 and 48 h after pMCAO. MMF treatment did not influence infarct size, however reduced edema volume at both 24 and 48 h after pMCAO. MMF treatment resulted in increased Hsp72 expression in the brain 6 h after pMCAO. Hcar2 mRNA levels increased significantly 24 h after pMCAO, but were not different between saline- and MMF-treated mice. MMF treatment also increased the level of the anti-inflammatory cytokine IL-10 in the brain and plasma 6 h after pMCAO, and additionally reduced the level of the pro-inflammatory cytokine IL-12p70 in the brain at 24 and 48 h after pMCAO. Conclusions A single intravenous bolus of MMF improved sensory-motor function after ischemic stroke, reduced edema formation, and increased the levels of the neuroprotective protein Hsp72 in the brain. The early increase in IL-10 and reduction in IL-12p70 in the brain combined with changes in systemic cytokine levels may also contribute to the functional recovery after pMCAO.

AB - Background Oxidative stress and inflammation exacerbate tissue damage in the brain after ischemic stroke. Dimethyl-fumarate (DMF) and its metabolite monomethyl-fumarate (MMF) are known to stimulate anti-oxidant pathways and modulate inflammatory responses. Considering these dual effects of fumarates, we examined the effect of MMF treatment after ischemic stroke in mice. Methods Permanent middle cerebral artery occlusion (pMCAO) was performed using adult, male C57BL/6 mice. Thirty minutes after pMCAO, 20 mg/kg MMF was administered intravenously. Outcomes were evaluated 6, 24 and 48 h after pMCAO. First, we examined whether a bolus of MMF was capable of changing expression of kelch-like erythroid cell-derived protein with CNC homology-associated protein 1 (Keap1) and nuclear factor erythroid 2-related factor (Nrf)2 in the infarcted brain. Next, we studied the effect of MMF on functional recovery. To explore mechanisms potentially influencing functional changes, we examined infarct volumes, edema formation, the expression of heat shock protein (Hsp)72, hydroxycarboxylic acid receptor 2 (Hcar2), and inducible nitric oxide synthase (iNOS) in the infarcted brain using real-time PCR and Western blotting. Concentrations of a panel of pro- and anti-inflammatory cytokines (IFNγ, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12p70, TNF) were examined in both the infarcted brain tissue and plasma samples 6, 24 and 48 h after pMCAO using multiplex electrochemoluminiscence analysis. Results Administration of MMF increased the protein level of Nrf2 6 h after pMCAO, and improved functional outcome at 24 and 48 h after pMCAO. MMF treatment did not influence infarct size, however reduced edema volume at both 24 and 48 h after pMCAO. MMF treatment resulted in increased Hsp72 expression in the brain 6 h after pMCAO. Hcar2 mRNA levels increased significantly 24 h after pMCAO, but were not different between saline- and MMF-treated mice. MMF treatment also increased the level of the anti-inflammatory cytokine IL-10 in the brain and plasma 6 h after pMCAO, and additionally reduced the level of the pro-inflammatory cytokine IL-12p70 in the brain at 24 and 48 h after pMCAO. Conclusions A single intravenous bolus of MMF improved sensory-motor function after ischemic stroke, reduced edema formation, and increased the levels of the neuroprotective protein Hsp72 in the brain. The early increase in IL-10 and reduction in IL-12p70 in the brain combined with changes in systemic cytokine levels may also contribute to the functional recovery after pMCAO.

KW - Cytokines

KW - Dimethyl-fumarate

KW - Focal cerebral ischemia

KW - Heat shock protein

KW - Monomethyl-fumarate

KW - Nrf2

KW - Tecfidera

KW - Brain Ischemia/drug therapy

KW - Kelch-Like ECH-Associated Protein 1/biosynthesis

KW - Stroke/complications

KW - Mice, Inbred C57BL

KW - NF-E2-Related Factor 2/biosynthesis

KW - Male

KW - Treatment Outcome

KW - Brain Edema/drug therapy

KW - Heat-Shock Proteins/biosynthesis

KW - Nitric Oxide Synthase Type II/antagonists & inhibitors

KW - Dimethyl Fumarate/therapeutic use

KW - Animals

KW - Behavior, Animal/drug effects

KW - Mice

KW - Infarction, Middle Cerebral Artery/drug therapy

KW - Cytokines/biosynthesis

U2 - 10.1016/j.expneurol.2017.06.011

DO - 10.1016/j.expneurol.2017.06.011

M3 - Journal article

C2 - 28602832

VL - 295

SP - 144

EP - 154

JO - Experimental Neurology

JF - Experimental Neurology

SN - 0014-4886

ER -