Neurochemical Research

, Volume 37, Issue 8, pp 1615–1623 | Cite as

Naringin Treatment Improves Functional Recovery by Increasing BDNF and VEGF Expression, Inhibiting Neuronal Apoptosis After Spinal Cord Injury

  • Wei Rong
  • Jun Wang
  • Xiaoguang Liu
  • Liang Jiang
  • Feng Wei
  • Xing Hu
  • Xiaoguang Han
  • Zhongjun Liu
Original Paper

Abstract

The aim of this study was to determine the therapeutic efficacy of starting naringin treatment 1 day after spinal cord injury (SCI) in rat and to investigate the underlying mechanism. SCI was induced using the modified weight-drop method in Sprague–Dawley rats. The SCI animals were randomly divided into three groups: vehicle-treated group; 20 mg/kg naringin-treated group; 40 mg/kg naringin-treated group, and additionally with sham group (laminectomy only). Locomotors functional recovery was assessed during the 6 weeks post operation period by performing open-field locomotors tests and inclined-plane tests. At the end of the study, the segments of spinal cord encompassing the injury site were removed for histopathological analysis. Immunohistochemistry was performed to observe the expression of the brain-derived neurotrophic factor (BDNF). The expression of vascular endothelial growth factor (VEGF), B-cell CLL/lymphoma-2 (Bcl-2), BCL-2-associated X protein (Bax) and caspase-3 were detected by Western blot analysis. The apoptotic neural cells were assessed using the TUNEL method. The results showed that the naringin-treated animals had significantly better locomotor function recovery, less myelin loss, and higher expression of BDNF and VEGF. In addition, naringin treatment significantly increased in Bcl-2:Bax ratio, reduced the enzyme activity of caspase-3 and decreased the number of apoptotic cells after SCI. These findings suggest that naringin treatment starting 1 day after SCI can significantly improve locomotor recovery, and this neuroprotective effect may be related to the upregulation of BDNF and VEGF and the inhibition of neural apoptosis. Therefore, naringin may be useful as a promising therapeutic agent for SCI.

Keywords

Spinal cord injury Naringin BDNF VEGF Apoptosis 

Notes

Acknowledgment

This work was supported by the Nature Science Foundation of China (NO: 58441-06).

References

  1. 1.
    Wang CX, Olschowka JA, Wrathall JR (1997) Increase of interleukin-1beta mRNA and protein in the spinal cord following experimental traumatic injury in the rat. Brain Res 759(2):190–196PubMedCrossRefGoogle Scholar
  2. 2.
    Oyinbo CA (2011) Secondary injury mechanisms in traumatic spinal cord injury: a nugget of this multiply cascade. Acta Neurobiol Exp 71(2):281–299Google Scholar
  3. 3.
    Blight AR (2002) Miracles and molecules—progress in spinal cord repair. Nat Neurosci 5 Suppl:1051–1054PubMedCrossRefGoogle Scholar
  4. 4.
    Yakovlev AG, Faden AI (2001) Caspase-dependent apoptotic pathways in CNS injury. Mol Neurobiol 24(1–3):131–144PubMedGoogle Scholar
  5. 5.
    Dougherty KD, Dreyfus CF, Black IB (2000) Brain-derived neurotrophic factor in astrocytes, oligodendrocytes, and microglia/macrophages after spinal cord injury. Neurobiol Dis 7(6 Pt B):574–585PubMedCrossRefGoogle Scholar
  6. 6.
    Nakajima H, Uchida K, Yayama T et al (2010) Targeted retrograde gene delivery of brain-derived neurotrophic factor suppresses apoptosis of neurons and oligodendroglia after spinal cord injury in rats. Spine (Phila Pa 1976) 35(5):497–504CrossRefGoogle Scholar
  7. 7.
    Greenberg DA, Jin K (2005) From angiogenesis to neuropathology. Nature 438(7070):954–959PubMedCrossRefGoogle Scholar
  8. 8.
    Pereira JE, Costa LM, Cabrita AM et al (2009) Methylprednisolone fails to improve functional and histological outcome following spinal cord injury in rats. Exp Neurol 220(1):71–81PubMedCrossRefGoogle Scholar
  9. 9.
    Zbarsky V, Datla KP, Parkar S et al (2005) Neuroprotective properties of the natural phenolic antioxidants curcumin and naringenin but not quercetin and fisetin in a 6-OHDA model of Parkinson’s disease. Free Radical Res 39(10):1119–1125CrossRefGoogle Scholar
  10. 10.
    Cavia-Saiz M, Busto MD, Pilar-Izquierdo MC et al (2010) Antioxidant properties, radical scavenging activity and biomolecule protection capacity of flavonoid naringenin and its glycoside naringin: a comparative study. J Sci Food Agric 90(7):1238–1244PubMedCrossRefGoogle Scholar
  11. 11.
    Anuja GI, Latha PG, Suja SR et al (2010) Anti-inflammatory and analgesic properties of Drynaria quercifolia (L.) J. Smith. J Ethnopharmacol 132(2):456–460PubMedCrossRefGoogle Scholar
  12. 12.
    Choe SC, Kim HS, Jeong TS et al (2001) Naringin has an antiatherogenic effect with the inhibition of intercellular adhesion molecule-1 in hypercholesterolemic rabbits. J Cardiovasc Pharmacol 38(6):947–955PubMedCrossRefGoogle Scholar
  13. 13.
    Gopinath K, Prakash D, Sudhandiran G (2011) Neuroprotective effect of naringin, a dietary flavonoid against 3-nitropropionic acid-induced neuronal apoptosis. Neurochem Int 59(7):1066–1073PubMedCrossRefGoogle Scholar
  14. 14.
    Butterfield DA, Howard B, Yatin S et al (1999) Elevated oxidative stress in models of normal brain aging and Alzheimer’s disease. Life Sci 65(18–19):1883–1892PubMedCrossRefGoogle Scholar
  15. 15.
    Gaur V, Aggarwal A, Kumar A (2009) Protective effect of naringin against ischemic reperfusion cerebral injury: possible neurobehavioral, biochemical and cellular alterations in rat brain. Eur J Pharmacol 616(1–3):147–154PubMedCrossRefGoogle Scholar
  16. 16.
    Golechha M, Chaudhry U, Bhatia J et al (2011) Naringin protects against kainic acid-induced status epilepticus in rats: evidence for an antioxidant, anti-inflammatory and neuroprotective intervention. Biol Pharm Bull 34(3):360–365PubMedCrossRefGoogle Scholar
  17. 17.
    Perot PL Jr, Lee WA, Hsu CY et al (1987) Therapeutic model for experimental spinal cord injury in the rat: I. Mortality and motor deficit. Cent Nerv Syst Trauma J Am Paralysis Assoc 4(3):149–159Google Scholar
  18. 18.
    Han X, Yang N, Xu Y et al (2011) Simvastatin treatment improves functional recovery after experimental spinal cord injury by upregulating the expression of BDNF and GDNF. Neurosci Lett 487(3):255–259PubMedCrossRefGoogle Scholar
  19. 19.
    Basso DM, Beattie MS, Bresnahan JC (1995) A sensitive and reliable locomotor rating scale for open field testing in rats. J Neurotrauma 12:1–21PubMedCrossRefGoogle Scholar
  20. 20.
    Fehlings MG, Tator CH (1992) The effect of direct current field polarity on recovery after acute experimental spinal cord injury. Brain Res 579(1):32–42PubMedCrossRefGoogle Scholar
  21. 21.
    Tyor WR, Avgeropoulos N, Ohlandt G et al (2002) Treatment of spinal cord impact injury in the rat with transforming growth factor-beta. J Neurol Sci 200(1–2):33–41PubMedCrossRefGoogle Scholar
  22. 22.
    Ray SK, Hogan EL, Banik NL (2003) Calpain in the pathophysiology of spinal cord injury: neuroprotection with calpain inhibitors. Brain Res Brain Res Rev 42(2):169–185PubMedCrossRefGoogle Scholar
  23. 23.
    Beattie MS (2004) Inflammation and apoptosis: linked therapeutic targets in spinal cord injury. Trends Mol Med 10(12):580–583PubMedCrossRefGoogle Scholar
  24. 24.
    Kanno S, Shouji A, Asou K et al (2003) Effects of naringin on hydrogen peroxide-induced cytotoxicity and apoptosis in P388 cells. J Pharmacol Sci 92(2):166–170PubMedCrossRefGoogle Scholar
  25. 25.
    Lu YH, Su MY, Huang HY et al (2010) Protective effects of the citrus flavanones to PC12 cells against cytotoxicity induced by hydrogen peroxide. Neurosci Lett 484(1):6–11PubMedCrossRefGoogle Scholar
  26. 26.
    Kumar A, Prakash A, Dogra S (2010) Naringin alleviates cognitive impairment, mitochondrial dysfunction and oxidative stress induced by D-galactose in mice. Food Chem Toxicol 48(2):626–632PubMedCrossRefGoogle Scholar
  27. 27.
    MacGregor JT (1986) Genetic toxicology of dietary flavonoids. Prog Clin Biol Res 206:33–43PubMedGoogle Scholar
  28. 28.
    Havsteen BH (2002) The biochemistry and medical significance of the flavonoids. Pharmacol Ther 96(2–3):67–202PubMedCrossRefGoogle Scholar
  29. 29.
    de Vries JH, Janssen PL, Hollman PC et al (1997) Consumption of quercetin and kaempferol in free-living subjects eating a variety of diets. Cancer Lett 114(1–2):141–144PubMedGoogle Scholar
  30. 30.
    Crowe MJ, Bresnahan JC, Shuman SL et al (1997) Apoptosis and delayed degeneration after spinal cord injury in rats and monkeys. Nat Med 3(2):73–76PubMedCrossRefGoogle Scholar
  31. 31.
    Widenfalk J, Lundstromer K, Jubran M et al (2001) Neurotrophic factors and receptors in the immature and adult spinal cord after mechanical injury or kainic acid. J Neurosci 21(10):3457–3475PubMedGoogle Scholar
  32. 32.
    Ray SK, Matzelle DD, Wilford GG et al (2000) Increased calpain expression is associated with apoptosis in rat spinal cord injury: calpain inhibitor provides neuroprotection. Neurochem Res 25(9):1191–1198PubMedCrossRefGoogle Scholar
  33. 33.
    Lu P, Jones LL, Tuszynski MH (2005) BDNF-expressing marrow stromal cells support extensive axonal growth at sites of spinal cord injury. Exp Neurol 191(2):344–360PubMedCrossRefGoogle Scholar
  34. 34.
    Lei Y, Fu W, Chen J et al (2011) Neuroprotective effects of Abacopterin E from Abacopteris penangiana against oxidative stress-induced neurotoxicity. J Ethnopharmacol 134(2):275–280PubMedCrossRefGoogle Scholar
  35. 35.
    Widenfalk J, Lipson A, Jubran M et al (2003) Vascular endothelial growth factor improves functional outcome and decreases secondary degeneration in experimental spinal cord contusion injury. Neuroscience 120(4):951–960PubMedCrossRefGoogle Scholar
  36. 36.
    Schindler R, Mentlein R (2006) Flavonoids and vitamin E reduce the release of the angiogenic peptide vascular endothelial growth factor from human tumor cells. J Nutr 136(6):1477–1482PubMedGoogle Scholar
  37. 37.
    Luo H, Jiang B-H, King SM et al (2008) Inhibition of cell growth and VEGF expression in ovarian cancer cells by flavonoids. Nutr Cancer Int J 60(6):800–809CrossRefGoogle Scholar
  38. 38.
    Kamat J, Devasagayam T (2000) Oxidative damage to mitochondria in normal and cancer tissues, and its modulation. Toxicology 155(1–3):73–82PubMedCrossRefGoogle Scholar
  39. 39.
    Borner C (2003) The Bcl-2 protein family: sensors and checkpoints for life-or-death decisions. Mol Immunol 39(11):615–647PubMedCrossRefGoogle Scholar
  40. 40.
    Chinnaiyan AM, Orth K, O’Rourke K et al (1996) Molecular ordering of the cell death pathway. Bcl-2 and Bcl-xL function upstream of the CED-3-like apoptotic proteases. J Biol Chem 271(9):4573–4576PubMedCrossRefGoogle Scholar
  41. 41.
    Springer JE, Azbill RD, Knapp PE (1999) Activation of the caspase-3 apoptotic cascade in traumatic spinal cord injury. Nat Med 5(8):943–946PubMedCrossRefGoogle Scholar
  42. 42.
    Choi BS, Sapkota K, Kim S et al (2010) Antioxidant activity and protective effects of Tripterygium regelii extract on hydrogen peroxide-induced injury in human dopaminergic cells, SH-SY5Y. Neurochem Res 35(8):1269–1280PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Wei Rong
    • 1
  • Jun Wang
    • 1
  • Xiaoguang Liu
    • 1
  • Liang Jiang
    • 1
  • Feng Wei
    • 1
  • Xing Hu
    • 1
  • Xiaoguang Han
    • 1
  • Zhongjun Liu
    • 1
  1. 1.Department of OrthopedicsPeking University Third HospitalBeijingChina

Personalised recommendations