Journal of Natural Medicines

, Volume 68, Issue 3, pp 586–603 | Cite as

Evaluation of the neuroprotective effect of chrysin via modulation of endogenous biomarkers in a rat model of spinal cord injury

  • Amit D. Kandhare
  • V. Shivakumar
  • Anuchandra Rajmane
  • Pinaki Ghosh
  • Subhash L. BodhankarEmail author
Original Paper


The objective of the present investigation was to evaluate the neuroprotective efficacy of chrysin in an experimental rat model of spinal cord injury (SCI). SCI was induced in male Sprague–Dawley rats by placing an aneurysm clip extradurally for 60 s at T10. The rats received treatment with either vehicle (SCI control) or chrysin (10, 20 and 40 mg/kg, p.o.) for 28 days. The various behavioral, biochemical and molecular parameters were determined. Chronic treatment with chrysin (20 and 40 mg/kg) significantly and dose-dependently (P < 0.05) attenuated the decrease in body weight, urine output, footprint analysis, sperm count and organ weight (testis, seminal vesicle and urinary bladder). It significantly improved (P < 0.05) the nociceptive threshold, motor and sensory nerve conduction velocity. The decreased activity of superoxide dismutase, reduced glutathione and membrane-bound inorganic phosphate were significantly (P < 0.05) restored by chrysin treatment. SCI resulted in a significant increase (P < 0.05) in lipid peroxidase, nitric oxide, tumor necrosis factor alpha, interleukin-1β, and bax whereas expression of bcl-2 and caspase-3 were significantly (P < 0.05) reduced. These changes were significantly reduced by treatment with chrysin (20 and 40 mg/kg, P < 0.05). Histological aberration induced after SCI in spinal cord, testis, kidney and urinary bladder were restored by treatment with chrysin (20 and 40 mg/kg). In conclusion, chrysin is a potential flavone-possessing antioxidant and its antiapoptotic property caused the subsequent recovery of both motor and sensory functions via modulation of endogenous biomarkers and neuronal apoptosis to inhibit the incidence of neurological deficits due to SCI.

Graphical Abstract


bax bcl-2 Caspase Chrysin Interleukin-1β Neuroprotective Spinal cord injury Tumor necrosis factor-α 



The authors would like acknowledge Dr. SS Kadam, Vice-Chancellor and Dr. KR Mahadik, Principal, Poona College of Pharmacy, Bharati Vidyapeeth Deemed University, Pune, India, for providing necessary facilities to carry out the study. We are also thankful to the All India Council of Technical and Education (AICTE), India for financial support by awarding GATE Scholarship to one of the authors, Mr. AD Kandhare for the research work.

Conflict of interest

There is no conflict of interest between any of the authors.

Supplementary material

11418_2014_840_MOESM1_ESM.docx (32 kb)
Supplementary material 1 (DOCX 32 kb)
11418_2014_840_MOESM2_ESM.pdf (167 kb)
Supplementary material 2 (PDF 167 kb)


  1. 1.
    Yucel N, Cayli SR, Ates O, Karadag N, Firat S, Turkoz Y (2006) Evaluation of the neuroprotective effects of citicoline after experimental spinal cord injury: improved behavioral and neuroanatomical recovery. Neurochem Res 31:767–775PubMedCrossRefGoogle Scholar
  2. 2.
    Tator CH, Fehlings MG (1991) Review of the secondary injury theory of acute spinal cord trauma with emphasis on vascular mechanisms. J Neurosurg 75:15–26PubMedCrossRefGoogle Scholar
  3. 3.
    Xu J, Hsu CY, Junker H, Chao S, Hogan EL, Chao J (1991) Kininogen and kinin in experimental spinal cord injury. J Neurochem 57:975–980PubMedCrossRefGoogle Scholar
  4. 4.
    Xing B, Chen H, Zhang M, Zhao D, Jiang R, Liu X et al (2008) Ischemic postconditioning inhibits apoptosis after focal cerebral ischemia/reperfusion injury in the rat. Stroke J Cereb Circ 39:2362–2369CrossRefGoogle Scholar
  5. 5.
    Ismailoglu O, Oral B, Gorgulu A, Sutcu R, Demir N (2010) Neuroprotective effects of tamoxifen on experimental spinal cord injury in rats. J Clin Neurosci 17:1306–1310PubMedCrossRefGoogle Scholar
  6. 6.
    Takahara Y, Maeda M, Nakatani T, Kiyama H (2007) Transient suppression of the vesicular acetylcholine transporter in urinary bladder pathways following spinal cord injury. Brain Res 1137:20–28PubMedCrossRefGoogle Scholar
  7. 7.
    Bartholdi D, Schwab ME (1997) Expression of pro-inflammatory cytokine and chemokine mRNA upon experimental spinal cord injury in mouse: an in situ hybridization study. Eur J Neurosci 9:1422–1438PubMedCrossRefGoogle Scholar
  8. 8.
    Bracken MB, Holford TR (1993) Effects of timing of methylprednisolone or naloxone administration on recovery of segmental and long-tract neurological function in NASCIS 2. J Neurosurg 79:500–507PubMedCrossRefGoogle Scholar
  9. 9.
    Kahraman S, Duz B, Kayali H, Korkmaz A, Oter S, Aydin A et al (2007) Effects of methylprednisolone and hyperbaric oxygen on oxidative status after experimental spinal cord injury: a comparative study in rats. Neurochem Res 32:1547–1551PubMedCrossRefGoogle Scholar
  10. 10.
    Sayer FT, Kronvall E, Nilsson OG (2006) Methylprednisolone treatment in acute spinal cord injury: the myth challenged through a structured analysis of published literature. Spine J 6:335–343PubMedCrossRefGoogle Scholar
  11. 11.
    Pietta PG (2000) Flavonoids as antioxidants. J Nat Prod 63:1035–1042PubMedCrossRefGoogle Scholar
  12. 12.
    Cardenas M, Marder M, Blank VC, Roguin LP (2006) Antitumor activity of some natural flavonoids and synthetic derivatives on various human and murine cancer cell lines. Bioorg Med Chem 14:2966–2971PubMedCrossRefGoogle Scholar
  13. 13.
    Lukacinova AMJ, Benacka R, Keller J, Maguth T, Kurila P et al (2008) Preventive effect of flavonoids on alloxan-induced diabetes mellitus in rats. Acta Vet Brno 77:175–182CrossRefGoogle Scholar
  14. 14.
    Cho H, Yun CW, Park WK, Kong JY, Kim KS, Park Y et al (2004) Modulation of the activity of pro-inflammatory enzymes, COX-2 and iNOS, by chrysin derivatives. Pharmacol Res 49:37–43PubMedCrossRefGoogle Scholar
  15. 15.
    Critchfield JW, Butera ST, Folks TM (1996) Inhibition of HIV activation in latently infected cells by flavonoid compounds. AIDS Res Hum Retrovir 12:39–46PubMedCrossRefGoogle Scholar
  16. 16.
    Pushpavalli G, Kalaiarasi P, Veeramani C, Pugalendi KV (2010) Effect of chrysin on hepatoprotective and antioxidant status in D-galactosamine-induced hepatitis in rats. Eur J Pharmacol 631:36–41PubMedCrossRefGoogle Scholar
  17. 17.
    Tahir M, Sultana S (2011) Chrysin modulates ethanol metabolism in Wistar rats: a promising role against organ toxicities. Alcohol Alcohol 46:383–392PubMedCrossRefGoogle Scholar
  18. 18.
    Gul S, Hanci V, Bahadir B, Acikgoz S, Bektas S, Ankarali H et al (2010) The effectiveness of dexmedetomidine in experimental spinal cord injury compared to methylprednisolone in rats. J Clin Neurosci 17:490–494PubMedCrossRefGoogle 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.
    Kandhare AD, Raygude KS, Ghosh P, Ghule AE, Bodhankar SL (2012) Neuroprotective effect of naringin by modulation of endogenous biomarkers in streptozotocin induced painful diabetic neuropathy. Fitoterapia 83:650–659PubMedCrossRefGoogle Scholar
  21. 21.
    Klapdor K, Dulfer BG, Hammann A, Van der Staay FJ (1997) A low-cost method to analyse footprint patterns. J Neurosci Methods 75:49–54PubMedCrossRefGoogle Scholar
  22. 22.
    Kandhare AD, Raygude KS, Shiva Kumar V, Rajmane AR, Visnagri A, Ghule AE, et al (2012) Ameliorative effects quercetin against impaired motor nerve function, inflammatory mediators and apoptosis in neonatal streptozotocin-induced diabetic neuropathy in rats. Biomed Aging Pathol 2:173–186Google Scholar
  23. 23.
    Misra HP, Fridovich I (1972) The role of superoxide anion in the autoxidation of epinephrine and a simple assay for superoxide dismutase. J Biol Chem 247:3170–3175PubMedGoogle Scholar
  24. 24.
    Moron MS, Depierre JW, Mannervik B (1979) Levels of glutathione, glutathione reductase and glutathione S-transferase activities in rat lung and liver. Biochim Biophys Acta 582:67–78PubMedCrossRefGoogle Scholar
  25. 25.
    Slater TF, Sawyer BC (1971) The stimulatory effects of carbon tetrachloride and other halogenoalkanes on peroxidative reactions in rat liver fractions in vitro. General features of the systems used. Biochem J 123:805–814PubMedCentralPubMedGoogle Scholar
  26. 26.
    Miranda KM, Espey MG, Wink DA (2001) A rapid, simple spectrophotometric method for simultaneous detection of nitrate and nitrite. Nitric Oxide 5:62–71PubMedCrossRefGoogle Scholar
  27. 27.
    Bonting (1970) Membrane and Ion transport. In: Bilter EE (ed) Presence of enzyme system in mammalian tissues. Wiley Inter Science, London pp 257–263Google Scholar
  28. 28.
    Kandhare AD, Ghosh P, Ghule AE, Bodhankar SL (2013) Elucidation of molecular mechanism involved in neuroprotective effect of Coenzyme Q10 in alcohol induced neuropathic pain. Fundam Clin Pharmacol 27:603–622PubMedCrossRefGoogle Scholar
  29. 29.
    Konturek PC, Duda A, Brzozowski T, Konturek SJ, Kwiecien S, Drozdowicz D et al (2000) Activation of genes for superoxide dismutase, interleukin-1beta, tumor necrosis factor-alpha, and intercellular adhesion molecule-1 during healing of ischemia-reperfusion-induced gastric injury. Scand J Gastroenterol 35:452–463PubMedCrossRefGoogle Scholar
  30. 30.
    Collins WF (1983) A review and update of experiment and clinical studies of spinal cord injury. Paraplegia 21:204–219PubMedCrossRefGoogle Scholar
  31. 31.
    Yune TY, Kim SJ, Lee SM, Lee YK, Oh YJ, Kim YC et al (2004) Systemic administration of 17beta-estradiol reduces apoptotic cell death and improves functional recovery following traumatic spinal cord injury in rats. J Neurotrauma 21:293–306PubMedCrossRefGoogle Scholar
  32. 32.
    Ji B, Li M, Budel S, Pepinsky RB, Walus L, Engber TM et al (2005) Effect of combined treatment with methylprednisolone and soluble Nogo-66 receptor after rat spinal cord injury. Eur J Neurosci 22:587–594PubMedCentralPubMedCrossRefGoogle Scholar
  33. 33.
    Santajuliana D, Hornfeldt BJ, Osborn JW (1996) Use of ganglionic blockers to assess neurogenic pressor activity in conscious rats. J Pharmacol Toxicol Methods 35:45–54PubMedCrossRefGoogle Scholar
  34. 34.
    Bain JR, Mackinnon SE, Hunter DA (1989) Functional evaluation of complete sciatic, peroneal, and posterior tibial nerve lesions in the rat. Plast Reconstr Surg 83:129–138PubMedCrossRefGoogle Scholar
  35. 35.
    Kasai M, Fukumitsu H, Soumiya H, Furukawa S (2011) Ethanol extract of chinese propolis facilitates functional recovery of locomotor activity after spinal cord injury. Evid Based Complement Altern Med 2011:9CrossRefGoogle Scholar
  36. 36.
    Raygude KS, Kandhare AD, Ghosh P, Ghule AE, Bodhankar SL (2012) Evaluation of ameliorative effect of quercetin in experimental model of alcoholic neuropathy in rats. Inflammopharmacology 1–11Google Scholar
  37. 37.
    Visnagri A, Kandhare AD, Shiva Kumar V, Rajmane AR, Mohammad A, Ghosh P, et al (2012) Elucidation of ameliorative effect of co-enzyme Q10 in streptozotocin-induced diabetic neuropathic perturbation by modulation of electrophysiological, biochemical and behavioral markers. Biomed Aging Pathol 2:157–172Google Scholar
  38. 38.
    Kandhare AD, Raygude KS, Ghosh P, Ghule AE, Bodhankar SL (2012) Therapeutic role of curcumin in prevention of biochemical and behavioral aberration induced by alcoholic neuropathy in laboratory animals. Neurosci Lett 511:18–22PubMedCrossRefGoogle Scholar
  39. 39.
    Hutchinson KJ, Gomez-Pinilla F, Crowe MJ, Ying Z, Basso DM (2004) Three exercise paradigms differentially improve sensory recovery after spinal cord contusion in rats. Brain 127:1403–1414PubMedCrossRefGoogle Scholar
  40. 40.
    Torrejais MM, Soares JC, Matheus SM, Cassel FD, Mello JM, Basso NA (2002) Histochemical and SEM evaluation of the neuromuscular junctions from alcoholic rats. Tissue Cell 34:117–123PubMedCrossRefGoogle Scholar
  41. 41.
    Cao Q, Zhang YP, Iannotti C, DeVries WH, Xu XM, Shields CB et al (2005) Functional and electrophysiological changes after graded traumatic spinal cord injury in adult rat. Exp Neurol 191(Suppl 1):S3–S16PubMedCrossRefGoogle Scholar
  42. 42.
    Kandhare AD, Kumar VS, Adil M, Rajmane AR, Ghosh P, Bodhankar SL (2012) Investigation of gastro protective activity of Xanthium strumarium L. by modulation of cellular and biochemical marker. Orient Pharm Exp Med 12:287–299CrossRefGoogle Scholar
  43. 43.
    Naik AK, Tandan SK, Dudhgaonkar SP, Jadhav SH, Kataria M, Prakash VR et al (2006) Role of oxidative stress in pathophysiology of peripheral neuropathy and modulation by N-acetyl-l-cysteine in rats. Eur J Pain 10:573–579PubMedCrossRefGoogle Scholar
  44. 44.
    Kandhare AD, Raygude KS, Ghosh P, Ghule AE, Gosavi TP, Badole SL et al (2012) Effect of hydroalcoholic extract of Hibiscus rosa sinensis Linn. leaves in experimental colitis in rats. Asian Pac J Trop Biomed 5:337–344CrossRefGoogle Scholar
  45. 45.
    Gosavi TP, Ghosh P, Kandhare AD, Kumar VS, Adil M, Rajmane AR et al (2012) Therapeutic effect of H. pylori nosode, a homeopathic preparation in healing of chronic H. pylori infected ulcers in laboratory animals. Asian Pac J Trop Dis 2:S603–S611CrossRefGoogle Scholar
  46. 46.
    Sathiavelu J, Senapathy GJ, Devaraj R, Namasivayam N (2009) Hepatoprotective effect of chrysin on prooxidant-antioxidant status during ethanol-induced toxicity in female albino rats. J Pharm Pharmacol 61:809–817PubMedCrossRefGoogle Scholar
  47. 47.
    Tator CH (1996) Experimental and clinical studies of the pathophysiology and management of acute spinal cord injury. J Spinal Cord Med 19:206–214PubMedGoogle Scholar
  48. 48.
    Cayli SR, Kocak A, Yilmaz U, Tekiner A, Erbil M, Ozturk C et al (2004) Effect of combined treatment with melatonin and methylprednisolone on neurological recovery after experimental spinal cord injury. Eur Spine J 13:724–732PubMedCentralPubMedCrossRefGoogle Scholar
  49. 49.
    Sultana S, Verma K, Khan R (2012) Nephroprotective efficacy of chrysin against cisplatin-induced toxicity via attenuation of oxidative stress. J Pharm Pharmacol 64:872–881PubMedCrossRefGoogle Scholar
  50. 50.
    Patil MVK, Kandhare AD, Bhise SD (2012) Anti-arthritic and anti-inflammatory activity of Xanthium srtumarium L. ethanolic extract in Freund’s complete adjuvant induced arthritis. Biomed Aging Pathol 2:6–15CrossRefGoogle Scholar
  51. 51.
    Anderson DK, Hall ED (1993) Pathophysiology of spinal cord trauma. Ann Emerg Med 22:987–992PubMedCrossRefGoogle Scholar
  52. 52.
    Hecker M, Preiss C, Klemm P, Busse R (1996) Inhibition by antioxidants of nitric oxide synthase expression in murine macrophages: role of nuclear factor kappa B and interferon regulatory factor 1. Br J Pharmacol 118:2178–2184PubMedCentralPubMedCrossRefGoogle Scholar
  53. 53.
    Huang HF, Li MT, Wang S, Wang G, Ottenweller JE (2003) Spinal cord contusion impairs sperm motility in the rat without disrupting spermatogenesis. J Androl 24:371–380PubMedGoogle Scholar
  54. 54.
    Mimata H, Satoh F, Tanigawa T, Nomura Y, Ogata J (1993) Changes of rat urinary bladder during acute phase of spinal cord injury. Urol Int 51:89–93PubMedCrossRefGoogle Scholar
  55. 55.
    Chang S, Mao ST, Hu SJ, Lin WC, Cheng CL (2000) Studies of detrusor-sphincter synergia and dyssynergia during micturition in rats via fractional Brownian motion. IEEE Trans Biomed Eng 47:1066–1073PubMedCrossRefGoogle Scholar
  56. 56.
    Guy M, Newall R, Borzomato J, Kalra PA, Price C (2009) Use of a first-line urine protein-to-creatinine ratio strip test on random urines to rule out proteinuria in patients with chronic kidney disease. Nephrol Dial Transpl 24:1189–1193CrossRefGoogle Scholar
  57. 57.
    Kim D, Sands JM, Klein JD (2003) Changes in renal medullary transport proteins during uncontrolled diabetes mellitus in rats. Am J Physiol Renal Physiol 285:F303–F309PubMedGoogle Scholar
  58. 58.
    Stemmermann GN, Weiss L et al (1950) A study of the germinal epithelium in male paraplegics. Am J Clin Pathol 20:24–34PubMedGoogle Scholar
  59. 59.
    Yakovlev AG, Ota K, Wang G, Movsesyan V, Bao WL, Yoshihara K et al (2001) Differential expression of apoptotic protease-activating factor-1 and caspase-3 genes and susceptibility to apoptosis during brain development and after traumatic brain injury. J Neurosci 21:7439–7446PubMedGoogle Scholar
  60. 60.
    Springer JE, Azbill RD, Knapp PE (1999) Activation of the caspase-3 apoptotic cascade in traumatic spinal cord injury. Nat Med 5:943–946PubMedCrossRefGoogle Scholar
  61. 61.
    Chen J, Graham SH, Nakayama M, Zhu RL, Jin K, Stetler RA et al (1997) Apoptosis repressor genes Bcl-2 and Bcl-x-long are expressed in the rat brain following global ischemia. J Cereb Blood Flow Metab 17:2–10PubMedCrossRefGoogle Scholar
  62. 62.
    Lee SM, Yune TY, Kim SJ, Park DW, Lee YK, Kim YC et al (2003) Minocycline reduces cell death and improves functional recovery after traumatic spinal cord injury in the rat. J Neurotrauma 20:1017–1027PubMedCrossRefGoogle Scholar
  63. 63.
    Popovich PG, Stokes BT, Whitacre CC (1996) Concept of autoimmunity following spinal cord injury: possible roles for T lymphocytes in the traumatized central nervous system. J Neurosci Res 45:349–363PubMedCrossRefGoogle Scholar
  64. 64.
    Hougee S, Sanders A, Faber J, Graus YM, van den Berg WB, Garssen J et al (2005) Decreased pro-inflammatory cytokine production by LPS-stimulated PBMC upon in vitro incubation with the flavonoids apigenin, luteolin or chrysin, due to selective elimination of monocytes/macrophages. Biochem Pharmacol 69:241–248PubMedCrossRefGoogle Scholar
  65. 65.
    He XL, Wang YH, Bi MG, Du GH (2012) Chrysin improves cognitive deficits and brain damage induced by chronic cerebral hypoperfusion in rats. Eur J Pharmacol 680:41–48PubMedCrossRefGoogle Scholar
  66. 66.
    Ha SK, Moon E, Kim SY (2010) Chrysin suppresses LPS-stimulated proinflammatory responses by blocking NF-kappaB and JNK activations in microglia cells. Neurosci Lett 485:143–147PubMedCrossRefGoogle Scholar
  67. 67.
    Maher P, Akaishi T, Abe K (2006) Flavonoid fisetin promotes ERK-dependent long-term potentiation and enhances memory. Proc Natl Acad Sci USA 103:16568–16573PubMedCentralPubMedCrossRefGoogle Scholar
  68. 68.
    Li YL, Li LT, Yu M, Wang YZ, Ge HY, Song CQ (2012) Beneficial effects of the herbal medicine Di Huang Yin Zi in patients with spinal cord injury: a randomized, placebo-controlled clinical study. J Int Med Res 40:1715–1724Google Scholar

Copyright information

© The Japanese Society of Pharmacognosy and Springer Japan 2014

Authors and Affiliations

  • Amit D. Kandhare
    • 1
  • V. Shivakumar
    • 1
  • Anuchandra Rajmane
    • 1
  • Pinaki Ghosh
    • 1
  • Subhash L. Bodhankar
    • 1
    Email author
  1. 1.Department of Pharmacology, Poona College of PharmacyBharati Vidyapeeth Deemed UniversityPuneIndia

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