Omega-3 fatty acid supplementation and reduction of traumatic axonal injury in a rodent head injury model

Laboratory investigation

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  • 1 Department of Neurosurgery, West Virginia University School of Medicine, Morgantown, West Virginia; and
  • 2 Inflammation Research Foundation, Marblehead, Massachusetts
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Object

Traumatic brain injury remains the most common cause of death in persons under 45 years of age in the Western world. Recent evidence from animal studies suggests that supplementation with omega-3 fatty acid (O3FA) (particularly eicosapentaenoic acid [EPA] and docosahexaenoic acid [DHA]) improves functional outcomes following focal neural injury. The purpose of this study is to determine the benefits of O3FA supplementation following diffuse axonal injury in rats.

Methods

Forty adult male Sprague-Dawley rats were used. Three groups of 10 rats were subjected to an impact acceleration injury and the remaining group underwent a sham-injury procedure (surgery, but no impact injury). Two of the groups subjected to the injury were supplemented with 10 or 40 mg/kg/day of O3FA; the third injured group served as an unsupplemented control group. The sham-injured rats likewise received no O3FA supplementation. Serum fatty acid levels were determined from the isolated plasma phospholipids prior to the injury and at the end of the 30 days of supplementation. After the animals had been killed, immunohistochemical analysis of brainstem white matter tracts was performed to assess the presence of β-amyloid precursor protein (APP), a marker of axonal injury. Immunohistochemical analyses of axonal injury mechanisms—including analysis for caspase-3, a marker of apoptosis; RMO-14, a marker of neurofilament compaction; and cytochrome c, a marker of mitochondrial injury—were performed.

Results

Dietary supplementation with a fish oil concentrate rich in EPA and DHA for 30 days resulted in significant increases in O3FA serum levels: 11.6% ± 4.9% over initial levels in the 10 mg/kg/day group and 30.7% ± 3.6% in the 40 mg/kg/day group. Immunohistochemical analysis revealed significantly (p < 0.05) decreased numbers of APP-positive axons in animals receiving O3FA supplementation: 7.7 ± 14.4 axons per mm2 in the 10 mg/kg/day group and 6.2 ± 11.4 axons per mm2 in the 40 mg/kg/day group, versus 182.2 ± 44.6 axons per mm2 in unsupplemented animals. Sham-injured animals had 4.1 ± 1.3 APP-positive axons per mm2. Similarly, immunohistochemical analysis of caspase-3 expression demonstrated significant (p < 0.05) reduction in animals receiving O3FA supplementation, 18.5 ± 28.3 axons per mm2 in the 10 mg/kg/day group and 13.8 ± 18.9 axons per mm2 in the 40 mg/kg/day group, versus 129.3 ± 49.1 axons per mm2 in unsupplemented animals.

Conclusions

Dietary supplementation with a fish oil concentrate rich in the O3FAs EPA and DHA increases serum levels of these same fatty acids in a dose-response effect. Omega-3 fatty acid supplementation significantly reduces the number of APP-positive axons at 30 days postinjury to levels similar to those in uninjured animals. Omega-3 fatty acids are safe, affordable, and readily available worldwide to potentially reduce the burden of traumatic brain injury.

Abbreviations used in this paper: AA = arachidonic acid; APP = amyloid precursor protein; COX = cyclooxygenase; DHA = docosahexaenoic acid; EPA = eicosapentaenoic acid; LOX = lipoxygenase; NGS = normal goat serum; O3FA = omega-3 fatty acid; PBS = phosphate-buffered saline; Sir2 = silent information 2; TBI = traumatic brain injury; TRAAK = TWIK-related arachidonic acid–stimulated potassium channel; TREK = TWIK-related potassium channel; TWIK = 2-pore weakly inward rectifying potassium channel.

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Contributor Notes

Address correspondence to: James D. Mills, M.D., P.O. Box 9183, West Virginia University Health Sciences Center, Department of Neurosurgery, Morgantown, West Virginia 26506. email: jmills@hsc.wvu.edu.

Please include this information when citing this paper: published online July 16, 2010; DOI: 10.3171/2010.5.JNS08914.

  • 1

    Bazan NG: Neuroprotectin D1 (NPD1): a DHA-derived mediator that protects brain and retina against cell injury-induced oxidative stress. Brain Pathol 15:159166, 2005

    • Search Google Scholar
    • Export Citation
  • 2

    Blondeau N, , Widmann C, , Lazdunski M, & Heurteaux C: Polyunsaturated fatty acids induce ischemic and epileptic tolerance. Neuroscience 109:231241, 2002

    • Search Google Scholar
    • Export Citation
  • 3

    Buki A, , Okonkwo DO, , Wang KK, & Povlishock JT: Cytochrome c release and caspase activation in traumatic axonal injury. J Neurosci 20:28252834, 2000

    • Search Google Scholar
    • Export Citation
  • 4

    Calder PC: Long-chain n-3 fatty acids and inflammation: potential application in surgical and trauma patients. Braz J Med Biol Res 36:433446, 2003

    • Search Google Scholar
    • Export Citation
  • 5

    Calderon F, & Kim HY: Docosahexaenoic acid promotes neurite growth in hippocampal neurons. J Neurochem 90:979988, 2004

  • 6

    Cao DH, , Xu JF, , Xue RH, , Zheng WF, & Liu ZL: Protective effect of chronic ethyl docosahexaenoate administration on brain injury in ischemic gerbils. Pharmacol Biochem Behav 79:651659, 2004

    • Search Google Scholar
    • Export Citation
  • 7

    Danthi SJ, , Enyeart JA, & Enyeart JJ: Modulation of native T-type calcium channels by omega-3 fatty acids. Biochem Biophys Res Commun 327:485493, 2005

    • Search Google Scholar
    • Export Citation
  • 8

    Doppenberg EM, , Choi SC, & Bullock R: Clinical trials in traumatic brain injury. What can we learn from previous studies?. Ann N Y Acad Sci 825:305322, 1997

    • Search Google Scholar
    • Export Citation
  • 9

    Dusart I, & Schwab ME: Secondary cell death and the inflammatory reaction after dorsal hemisection of the rat spinal cord. Eur J Neurosci 6:712724, 1994

    • Search Google Scholar
    • Export Citation
  • 10

    Eldadah BA, & Faden AI: Caspase pathways, neuronal apoptosis, and CNS injury. J Neurotrauma 17:811829, 2000

  • 11

    Emsley R, , Oosthuizen P, & van Rensburg SJ: Clinical potential of omega-3 fatty acids in the treatment of schizophrenia. CNS Drugs 17:10811091, 2003

    • Search Google Scholar
    • Export Citation
  • 12

    Högyes E, , Nyakas C, , Kiliaan A, , Farkas T, , Penke B, & Luiten PG: Neuroprotective effect of developmental docosahexaenoic acid supplement against excitotoxic brain damage in infant rats. Neuroscience 119:9991012, 2003

    • Search Google Scholar
    • Export Citation
  • 13

    Holub BJ, & Skeaff CM: Nutritional regulation of cellular phosphatidylinositol. Methods Enzymol 141:234244, 1987

  • 14

    Jump DB: Dietary polyunsaturated fatty acids and regulation of gene transcription. Curr Opin Lipidol 13:155164, 2002

  • 15

    Lang-Lazdunski L, , Blondeau N, , Jarretou G, , Lazdunski M, & Heurteaux C: Linolenic acid prevents neuronal cell death and paraplegia after transient spinal cord ischemia in rats. J Vasc Surg 38:564575, 2003

    • Search Google Scholar
    • Export Citation
  • 16

    Lauritzen I, , Blondeau N, , Heurteaux C, , Widmann C, , Romey G, & Lazdunski M: Polyunsaturated fatty acids are potent neuroprotectors. EMBO J 19:17841793, 2000

    • Search Google Scholar
    • Export Citation
  • 17

    Lonergan PE, , Martin DS, , Horrobin DF, & Lynch MA: Neuroprotective actions of eicosapentaenoic acid on lipopolysaccharide-induced dysfunction in rat hippocampus. J Neurochem 91:2029, 2004

    • Search Google Scholar
    • Export Citation
  • 18

    Marmarou A, , Foda MA, , van den Brink W, , Campbell J, , Kita H, & Demetriadou K: A new model of diffuse brain injury in rats. Part I: Pathophysiology and biomechanics. J Neurosurg 80:22912300, 1994

    • Search Google Scholar
    • Export Citation
  • 19

    Mills JD, , Stone JR, , Rubin DG, , Melon DE, , Okonkwo DO, & Periasamy A, : Illuminating protein interactions in tissue using confocal and two-photon excitation fluorescent resonance energy transfer microscopy. J Biomed Optics 8:347356, 2003

    • Search Google Scholar
    • Export Citation
  • 20

    Mukherjee PK, , Marcheselli VL, , Serhan CN, & Bazan NG: Neuroprotectin D1: a docosahexaenoic acid-derived docosatriene protects human retinal pigment epithelial cells from oxidative stress. Proc Natl Acad Sci U S A 101:84918496, 2004

    • Search Google Scholar
    • Export Citation
  • 21

    Murphy EJ, , Behrmann D, , Bates CM, & Horrocks LA: Lipid alterations following impact spinal cord injury in the rat. Mol Chem Neuropathol 23:1326, 1994

    • Search Google Scholar
    • Export Citation
  • 22

    Nakagawa Y, , Nakamura M, , McIntosh TK, , Rodriguez A, , Berlin JA, & Smith DH, : Traumatic brain injury in young, amyloid-beta peptide overexpressing transgenic mice induces marked ipsilateral hippocampal atrophy and diminished Abeta deposition during aging. J Comp Neurol 411:390398, 1999

    • Search Google Scholar
    • Export Citation
  • 23

    Needleman P, , Raz A, , Minkes MS, , Ferrendelli JA, & Sprecher H: Triene prostaglandins: prostacyclin and thromboxane biosynthesis and unique biological properties. Proc Natl Acad Sci U S A 76:944948, 1979

    • Search Google Scholar
    • Export Citation
  • 24

    Raghupathi R, , Graham DI, & McIntosh TK: Apoptosis after traumatic brain injury. J Neurotrauma 17:927938, 2000

  • 25

    Roberts L, , Bailes J, , Dedhia H, , Zikos A, , Singh A, & McDowell D, : Surviving a mine explosion. J Am Coll Surg 207:276283, 2008

  • 26

    Sarsilmaz M, , Songur A, , Kus I, , Ozyurt B, , Gulec M, & Sogut S, : The regulatory role of dietary omega-3 essential fatty acids on oxidant/antioxidant balance in the rat hippocampus. Neurosci Res Commun 33:114123, 2003

    • Search Google Scholar
    • Export Citation
  • 27

    Schlanger S, , Shinitzky M, & Yam D: Diet enriched with omega-3 fatty acids alleviates convulsion symptoms in epilepsy patients. Epilepsia 43:103104, 2002

    • Search Google Scholar
    • Export Citation
  • 28

    Schwab JM, , Chiang N, , Arita M, & Serhan CN: Resolvin E1 and protectin D1 activate inflammation-resolution programmes. Nature 447:869874, 2007

    • Search Google Scholar
    • Export Citation
  • 29

    Songur A, , Sarsilmaz M, , Sogut S, , Ozyurt B, , Ozyurt H, & Zararsiz I, : Hypothalamic superoxide dismutase, xanthine oxidase, nitric oxide, and malondialdehyde in rats fed with fish omega-3 fatty acids. Prog Neuropsychopharmacol Biol Psychiatry 28:693698, 2004

    • Search Google Scholar
    • Export Citation
  • 30

    Sosin DM, , Sniezek JE, & Waxweiler RJ: Trends in death associated with traumatic brain injury, 1979 through 1992. Success and failure. JAMA 273:17781780, 1995

    • Search Google Scholar
    • Export Citation
  • 31

    Stone JR, , Walker SA, & Povlishock JT: The visualization of a new class of traumatically injured axons through the use of a modified method of microwave antigen retrieval. Acta Neuropathol 97:335345, 1999

    • Search Google Scholar
    • Export Citation
  • 32

    Sullivan PG, , Rabchevsky AG, , Waldmeier PC, & Springer JE: Mitochondrial permeability transition in CNS trauma: cause or effect of neuronal cell death?. J Neurosci Res 79:231239, 2005

    • Search Google Scholar
    • Export Citation
  • 33

    Wang HG, , Pathan N, , Ethell IM, , Krajewski S, , Yamaguchi Y, & Shibasaki F, : Ca2+-induced apoptosis through calcineurin dephosphorylation of BAD. Science 284:339343, 1999

    • Search Google Scholar
    • Export Citation
  • 34

    Wang X, , Zhao X, , Mao ZY, , Wang XM, & Liu ZL: Neuroprotective effect of docosahexaenoic acid on glutamate-induced cytotoxicity in rat hippocampal cultures. Neuroreport 14:2457 2461, 2003

    • Search Google Scholar
    • Export Citation
  • 35

    Waxweiler RJ, , Thurman D, , Sniezek J, , Sosin D, & O'Neil J: Monitoring the impact of traumatic brain injury: a review and update. J Neurotrauma 12:509516, 1995

    • Search Google Scholar
    • Export Citation
  • 36

    Wu A, , Ying Z, & Gomez-Pinilla F: Dietary omega-3 fatty acids normalize BDNF levels, reduce oxidative damage, and counteract learning disability after traumatic brain injury in rats. J Neurotrauma 21:14571467, 2004

    • Search Google Scholar
    • Export Citation
  • 37

    Wu A, , Ying Z, & Gomez-Pinilla F: Omega-3 fatty acids supplementation restores mechanisms that maintain brain homeostasis in traumatic brain injury. J Neurotrauma 24:15871595, 2007

    • Search Google Scholar
    • Export Citation
  • 38

    Xu GY, , Hughes MG, , Ye Z, , Hulsebosch CE, & McAdoo DJ: Concentrations of glutamate released following spinal cord injury kill oligodendrocytes in the spinal cord. Exp Neurol 187:329336, 2004

    • Search Google Scholar
    • Export Citation

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