Effect of Trehalose on Neurocan and Neural-Glial Antigen 2 Genes Expression in Rats with Spinal Cord Injury

Document Type: Original Article


1 Master of Science, Department of Clinical Biochemistry, Afzalipour School of Medicine, Kerman University of Medical Sciences, Kerman, Iran

2 Professor, Endocrinology and Metabolism Research Center, Institute of Basic and Clinical Physiology Sciences, Kerman University of Medical Sciences, Kerman, Iran

3 Assistant Professor, Neuroscience Research Center, Institute of Neuropharmacology, Department of Clinical Biochemistry, Afzalipour School of Medicine, Kerman University of Medical Sciences, Kerman, Iran


Background: Chondroitin sulfate proteoglycans (CSPGs) are the major cause of axonal regeneration failure at the site of lesion in spinal cord injury (SCI). Inflammation is believed to stimulate the upregulation of CSPGs expression. Recent evidence showed that trehalose reduces the development of inflammation in SCI. The aim of this study was to investigate the effect of trehalose on neurocan and Neural-Glial Antigen 2 (NG2) mRNA levels in SCI in rats.
Methods: In this experimental study, male rats were divided into six groups (n=15). Sham (laminectomy), SCI (laminectomy and SCI), vehicle (laminectomy and SCI, treated with phosphate buffer saline), and T10, T100 and T1000 (laminectomy and SCI, treated with 10, 100 and 1000 mM trehalose). Five rats in each group were sacrificed at 1, 3 and 7 days post-injury to measure neurocan and NG2 mRNA levels in lesion. Statistical analysis was performed using Kruskal-Wallis methods followed by the Mann-Whitney test.
Results: Findings indicated that SCI upregulated neurocan and NG2 mRNA levels at all times. No significant difference was observed in neurocan and NG2 gene transcripts between SCI and vehicle groups (p>0.05). However, 10 mM trehalose downregulated the mRNA level of both neurocan (0.76 and 0.65 fold) and NG2 (0.75 and 0.70 fold) at 3 and 7 days post-SCI compared to vehicle group (p p<0.01, respectively).
Conclusion: Collectively, treatment with low dose trehalose showed a decrease in neurocan and NG2 mRNA levels in spinal cord injured rats.


  1. Krityakiarana W, Zhao PM, Nguyen K, Gomez-Pinilla F, Kotchabhakdi N, de Vellis J, et al. Proof-of concept that an acute trophic factors intervention after spinal cord injury provides an adequate niche for neuroprotection, recruitment of nestin-expressing progenitors and regeneration. Neurochem Res 2016; 41(1-2):431-49.
  2. Martirosyan NL, Carotenuto A, Patel AA, Kalani MY, Yagmurlu K, Lemole Jr GM, et al. The role of microrna markers in the diagnosis, treatment, and outcome prediction of spinal cord injury. Front Surg 2016; 3:56.
  3. Anwar MA, Al Shehabi TS, Eid AH. Inflammogenesis of secondary spinal cord injury. Front Cell Neurosci 2016; 10:98.
  4. Janzadeh A, Sarveazad A, Yousefifard M, Dameni S, Samani FS, Mokhtarian K, et al. Combine effect of chondroitinase ABC and low level laser (660 nm) on spinal cord injury model in adult male rats. Neuropeptides 2017; 65:90-9.
  5. Dyck SM, Karimi-Abdolrezaee S. Chondroitin sulfate proteoglycans: key modulators in the developing and pathologic central nervous system. Exp Neurol 2015; 269:169-87.
  6. Cheng CH, Lin CT, Lee MJ, Tsai MJ, Huang WH, Huang MC, et al. Local delivery of high-dose chondroitinase ABC in the sub-acute stage promotes axonal outgrowth and functional recovery after complete spinal cord transection. PloS One 2015; 10(9):e0138705.
  7. Sherman LS, Back SA. A ‘GAG’reflex prevents repair of the damaged CNS. Trends Neurosci 2008; 31(1):44-52.
  8. Kwon BK, Tetzlaff W, Grauer JN, Beiner J, Vaccaro AR. Pathophysiology and pharmacologic treatment of acute spinal cord injury. Spine J 2004; 4(4):451-64.
  9. Bracken MB, Shepard MJ, Collins WF, Holford TR, Young W, Baskin DS, et al. A randomized, controlled trial of methylprednisolone or naloxone in the treatment of acute spinal-cord injury. results of the second national acute spinal cord injury study. N Engl J Med 1990; 322(20):1405-11.
  10. Bartholdi D, Schwab ME. Methylprednisolone inhibits early inflammatory processes but not ischemic cell death after experimental spinal cord lesion in the rat. Brain Res 1995; 672(1):177-86.
  11. Liu WL, Lee YH, Tsai SY, Hsu CY, Sun YY, Yang LY, et al. Methylprednisolone inhibits the expression of glial fibrillary acidic protein and chondroitin sulfate proteoglycans in reactivated astrocytes. Glia 2008; 56(13):1390-400.
  12. Holmberg E, Zhang SX, Sarmiere PD, Kluge BR, White JT, Doolen S. Statins decrease chondroitin sulfate proteoglycan expression and acute astrocyte activation in central nervous system injury. Exp Neurol 2008; 214(1):78-86.
  13. Echigo R, Shimohata N, Karatsu K, Yano F, Kayasuga-Kariya Y, Fujisawa A, et al. Trehalose treatment suppresses inflammation, oxidative stress, and vasospasm induced by experimental subarachnoid hemorrhage. J Transl Med 2012; 10:80.
  14. He Q, Wang Y, Lin W, Zhang Q, Zhao J, Liu FT, et al. Trehalose alleviates PC12 neuronal death mediated by lipopolysaccharide-stimulated BV-2 cells via inhibiting nuclear transcription factor NF-κB and AP-1 activation. Neurotox Res 2014; 26(4):430-9.
  15. Nazari Robati M, Akbari M, Khaksari M, Mirzaee M. Trehalose attenuates spinal cord injury through the regulation of oxidative stress, inflammation and GFAP expression in rats. J Spinal Cord Med 2018; 4:1-8.
  16. Akbari M, Khaksari M, Rezaeezadeh Roukerd M, Mirzaee M, Nazari Robati M. Effect of chondroitinase ABC on inflammatory and oxidative response following spinal cord injury. Iran J Basic Med Sci 2017; 20(7):806-12.
  17. Mirzaie M, Karimi M, Fallah H, Khaksari M, Nazari Robati M. Downregulation of matrix metalloproteinases 2 and 9 is involved in the protective effect of trehalose on spinal cord injury. Int J Mol Cel Med 2018; 7(1):8-16.
  18. Lee S, Zhao X, Hatch M, Chun S, Chang E. Central neuropathic pain in spinal cord injury. Crit Rev Phys Rehabil Med 2013; 25(3-4):159-72.
  19. Jones LL, Yamaguchi Y, Stallcup WB, Tuszynski MH. NG2 is a major chondroitin sulfate proteoglycan produced after spinal cord injury and is expressed by macrophages and oligodendrocyte progenitors. J Neurosci 2002; 22(7):2792-803.
  20. Iaci JF, Vecchione AM, Zimber MP, Caggiano AO. Chondroitin sulfate proteoglycans in spinal cord contusion injury and the effects of chondroitinase treatment. J Neurotrauma 2007; 24(11):1743-60.
  21. Massey JM, Amps J, Viapiano MS, Matthews RT, Wagoner MR, Whitaker CM, et al. Increased chondroitin sulfate proteoglycan expression in denervated brainstem targets following spinal cord injury creates a barrier to axonal regeneration overcome by chondroitinase ABC and neurotrophin-3. Exp Neurol 2008; 209(2):426-45.
  22. Jones LL, Sajed D, Tuszynski MH. Axonal regeneration through regions of chondroitin sulfate proteoglycan deposition after spinal cord injury: a balance of permissiveness and inhibition. J Neurosci 2003; 23(28):9276-88.
  23. Harris NG, Carmichael ST, Hovda DA, Sutton RL. Traumatic brain injury results in disparate regions of chondroitin sulfate proteoglycan expression that are temporally limited. J Neurosci Res 2009; 87(13):2937-50.
  24. Zehendner CM, Sebastiani A, Hugonnet A, Bischoff F, Luhmann HJ, Thal SC. Traumatic brain injury results in rapid pericyte loss followed by reactive pericytosis in the cerebral cortex. Sci Rep 2015; 5:13497
  25. Huang L, Wu ZB, Zhuge Q, Zheng W, Shao B, Wang B, et al. Glial scar formation occurs in the human brain after ischemic stroke. Int J Med Sci 2014; 11(4):344-8.
  26. Carmichael ST, Archibeque I, Luke L, Nolan T, Momiy J, Li S. Growth-associated gene expression after stroke: evidence for a growth-promoting region in peri-infarct cortex. Exp Neurol 2005; 193(2):291-311.
  27. Ambrosini E, Aloisi F. Chemokines and glial cells: a complex network in the central nervous system. Neurochem Res 2004; 29(5):1017-38.
  28. Jain NK, Roy I. Effect of trehalose on protein structure. Protein Sci 2009; 18(1):24-36. 
  29. Taya K, Hirose K, Hamada S. Trehalose inhibits inflammatory cytokine production by protecting IκB-α reduction in mouse peritoneal macrophages. Arch Oral Biol 2009; 54(8):749-56.
  30. Ampofo E, Schmitt BM, Menger MD, Laschke MW. The regulatory mechanisms of NG2/CSPG4 expression. Cell Mol Biol Lett 2017; 22:4.
  31. Fujiyoshi T, Kubo T, Chan CC, Koda M, Okawa A, Takahashi K. Interferon-γ decreases chondroitin sulfate proteoglycan expression and enhances hindlimb function after spinal cord injury in mice. J Neurotrauma 2010; 27(12):2283-94.
  32. Akbari M, Dabiri S, Nematollahi-Mahani SN, Nazari Robati M. Effect of enzyme chondroitinase ABC on glial fibrillary acidic protein, chondroitin sulfated proteoglycans and chondroitin 4-sulfate level in an animal model of spinal cord injury. J Kerman Univ Med Sci 2017; 24(4):259-67.