J Korean Neurosurg Soc.  2014 Nov;56(5):375-382. 10.3340/jkns.2014.56.5.375.

Intra-Spinal Bone Marrow Mononuclear Cells Transplantation Inhibits the Expression of Nuclear Factor-kappaB in Acute Transection Spinal Cord Injury in Rats

Affiliations
  • 1Department of Orthopaedics, First Affiliated Hospital of Jiamusi University, Heilongjiang, China. jianhua01@163.com

Abstract


OBJECTIVE
To assess the effect of bone marrow mononuclear cells (BMMNCs) transplantation in the expression of nuclear factor-kappaB (NF-kappaB) in spinal cord injury (SCI) in rats.
METHODS
BMMNCs were isolated from tibia and femur by a density gradient centrifugation. After establishment of acute transection SCI, rats were divided into experiment (BMMNCs), experiment control (0.1 M PBS infused) and sham surgery groups (laminectomy without any SCI). Locomotor function was assessed weekly for 5 weeks post-injury using BBB locomotor score and urinary bladder function daily for 4 weeks post-injury. Activity of NF-kappaB in spinal cord was assessed by immunohistochemistry and reverse transcriptase polymerase chain reaction.
RESULTS
At each time point post-injury, sham surgery group had significantly higher Basso, Beattie, Bresnahan locomotor and urinary bladder function scores than experiment and experiment control group (p<0.05). At subsequent time interval there were gradual improvement in both experiment and experiment control group, but experiment group had higher score in comparison to experiment control group (p<0.05). Comparisons were also made for expression of activated NF-kappaB positive cells and level of NF-kappaB messenger RNA in spinal cord at various time points between the groups. Activated NF-kappaB immunoreactivity and level of NF-kappaB mRNA expression were significantly higher in control group in comparison to experiment and sham surgery group (p<0.05).
CONCLUSION
BMMNCs transplantation attenuates the expression of NF-kappaB in injured spinal cord tissue and thus helps in recovery of neurological function in rat models with SCI.

Keyword

Cell transplantation; Inflammation; Mononuclear cells; Neuroprotective agent; NF-kappaB; Spinal cord injury

MeSH Terms

Animals
Bone Marrow*
Cell Transplantation
Centrifugation, Density Gradient
Femur
Immunohistochemistry
Inflammation
Models, Animal
NF-kappa B
Rats*
Reverse Transcriptase Polymerase Chain Reaction
RNA, Messenger
Spinal Cord
Spinal Cord Injuries*
Tibia
Urinary Bladder
NF-kappa B
RNA, Messenger

Figure

  • Fig. 1 Total numbers of activated NF-κB positive cells in spinal cord of rats at different time post-injury. All bars represent mean values and error bars represent standard deviation. There were significantly increased activated NF-κB positive cells in experiment and experiment control group in comparison to sham surgery group (p<0.05) and significant decrease in experiment group in comparison to experimental control group (p<0.05). *Re-presents the time point of highest and significant difference in expression of NF-κB post-injury. NF-κB : nuclear factor-κB.

  • Fig. 2 A : Immunohistochemical analysis for the expression of NF-κB in experimental group at 6 hours post-injury. B : Immunohistochemical analysis for the expression of NF-κB in control group 6 hours post-injury. C : Immunohistochemical analysis for the expression of NF-κB in Experimental group at 1 day post-injury. D : Immunohistochemical analysis for the expression of NF-κB in control group at 1 day post-injury. E : Immunohistochemical analysis for the expression of NF-κB in Experiment group at 3 days post-injury. F : Immunohistochemical analysis for the expression of NF-κB in Control group at 3 days post-injury. G : Immunohistochemical analysis for the expression of NF-κB in experimental group at 7 days post-injury. H : Immunohistochemical analysis for the expression of NF-κB in control group at 7 days post-injury. I : Immunohistochemical analysis for the expression of NF-κB in experimental group at 14 days post-injury. J : Immunohistochemical analysis for the expression of NF-κB in control group at 14 days post-injury. K : Immunohistochemical analysis for the expression of NF-κB in Sham surgery group at 6 hours post-injury. L : Immunohistochemical analysis for the expression of NF-κB in Sham surgery group at14 days post injury. NF-κB : nuclear factor-κB.

  • Fig. 3 Level of NF-κB mRNA expression in spinal cord of rats at different time post-injury. All bars represent mean values and error bars represent standard deviation. There were significantly increase in NF-κB mRNA level in experiment and experiment control group in comparison to sham surgery group (p<0.05) and significant decrease in experiment group in comparison to experimental control group (p<0.05). *Represents the time point of highest and significant difference in expression of NF-κB post-injury. NF-κB : nuclear factor-κB, mRNA : messenger RNA.

  • Fig. 4 A : Reverse transcription polymerase chain reaction (RT PCR) analysis of nuclear factor-κB expression in spinal cord (3 day post surgery). B : RT PCR analysis of GADPH expression in spinal cord (3 day post surgery). Lane 1 : marker, Lane 2 : Sham surgery group, Lane 3 : Experimental Control group, Lane 4 : Experimental group.

  • Fig. 5 Open field locomotor assessment by BBB score in rats at different time post-injury. All bars represent mean values and error bars represent standard deviation. There were significantly higher BBB score in rats from experiment group in comparison to experimental control group (p<0.05). BBB : Basso, Beattie, Bresnahan.

  • Fig. 6 Martin Schwab's Lab urinary bladder function score in rats at different time post-injury. All bars represent mean values and error bars represent standard deviation. There were significant urinary bladder function improvement in rats from experiment group in comparison to experimental control group (p<0.05).


Reference

1. Baeuerle PA. The inducible transcription activator NF-kappa B : regulation by distinct protein subunits. Biochim Biophys Acta. 1991; 1072:63–80. PMID: 2018779.
Article
2. Basso DM, Beattie MS, Bresnahan JC. A sensitive and reliable locomotor rating scale for open field testing in rats. J Neurotrauma. 1995; 12:1–21. PMID: 7783230.
Article
3. Beattie MS. Inflammation and apoptosis : linked therapeutic targets in spinal cord injury. Trends Mol Med. 2004; 10:580–583. PMID: 15567326.
Article
4. Bethea JR, Castro M, Keane RW, Lee TT, Dietrich WD, Yezierski RP. Traumatic spinal cord injury induces nuclear factor-kappaB activation. J Neurosci. 1998; 18:3251–3260. PMID: 9547234.
5. Brambilla R, Bracchi-Ricard V, Hu WH, Frydel B, Bramwell A, Karmally S, et al. Inhibition of astroglial nuclear factor kappaB reduces inflammation and improves functional recovery after spinal cord injury. J Exp Med. 2005; 202:145–156. PMID: 15998793.
Article
6. Brambilla R, Hurtado A, Persaud T, Esham K, Pearse DD, Oudega M, et al. Transgenic inhibition of astroglial NF-kappa B leads to increased axonal sparing and sprouting following spinal cord injury. J Neurochem. 2009; 110:765–778. PMID: 19522780.
Article
7. Cao HQ, Dong ED. An update on spinal cord injury research. Neurosci Bull. 2013; 29:94–102. PMID: 23124646.
Article
8. Chen LF, Greene WC. Shaping the nuclear action of NF-kappaB. Nat Rev Mol Cell Biol. 2004; 5:392–401. PMID: 15122352.
9. Esposito E, Paterniti I, Mazzon E, Genovese T, Galuppo M, Meli R, et al. MK801 attenuates secondary injury in a mouse experimental compression model of spinal cord trauma. BMC Neurosci. 2011; 12:31. PMID: 21492450.
Article
10. Genovese T, Paterniti I, Mazzon E, Esposito E, Di Paola R, Galuppo M, et al. Efficacy of treatment with verbascoside, biotechnologically produced by Syringa vulgaris plant cell cultures in an experimental mice model of spinal cord trauma. Naunyn Schmiedebergs Arch Pharmacol. 2010; 382:331–345. PMID: 20799028.
Article
11. Glazova M, Pak ES, Moretto J, Hollis S, Brewer KL, Murashov AK. Pre-differentiated embryonic stem cells promote neuronal regeneration by cross-coupling of BDNF and IL-6 signaling pathways in the host tissue. J Neurotrauma. 2009; 26:1029–1042. PMID: 19138107.
Article
12. Hamada Y, Ikata T, Katoh S, Tsuchiya K, Niwa M, Tsutsumishita Y, et al. Roles of nitric oxide in compression injury of rat spinal cord. Free Radic Biol Med. 1996; 20:1–9. PMID: 8903674.
Article
13. Han X, Lu M, Wang S, Lv D, Liu H. Targeting IKK/NF-κB pathway reduces infiltration of inflammatory cells and apoptosis after spinal cord injury in rats. Neurosci Lett. 2012; 511:28–32. PMID: 22289688.
Article
14. Han X, Wang S, Zhang Z, Lü D, Liu H. BMS-345541 inhibited nuclear factor kappa B expression and improved locomotor function recovery in rats after acute spinal cord injury. Neural Regen Res. 2011; 6:1775–1779.
15. Hausmann ON. Post-traumatic inflammation following spinal cord injury. Spinal Cord. 2003; 41:369–378. PMID: 12815368.
Article
16. Jia Z, Zhu H, Li J, Wang X, Misra H, Li Y. Oxidative stress in spinal cord injury and antioxidant-based intervention. Spinal Cord. 2012; 50:264–274. PMID: 21987065.
Article
17. Kerr BJ, Girolami EI, Ghasemlou N, Jeong SY, David S. The protective effects of 15-deoxy-delta-(12,14)-prostaglandin J2 in spinal cord injury. Glia. 2008; 56:436–448. PMID: 18205174.
Article
18. Kim GM, Xu J, Xu J, Song SK, Yan P, Ku G, et al. Tumor necrosis factor receptor deletion reduces nuclear factor-kappaB activation, cellular inhibitor of apoptosis protein 2 expression, and functional recovery after traumatic spinal cord injury. J Neurosci. 2001; 21:6617–6625. PMID: 11517251.
Article
19. La Rosa G, Cardali S, Genovese T, Conti A, Di Paola R, La Torre D, et al. Inhibition of the nuclear factor-kappaB activation with pyrrolidine dithiocarbamate attenuating inflammation and oxidative stress after experimental spinal cord trauma in rats. J Neurosurg Spine. 2004; 1:311–321. PMID: 15478370.
Article
20. Lee KM, Jeon SM, Cho HJ. Tumor necrosis factor receptor 1 induces interleukin-6 upregulation through NF-kappaB in a rat neuropathic pain model. Eur J Pain. 2009; 13:794–806. PMID: 18938092.
Article
21. Liebscher T, Schnell L, Schnell D, Scholl J, Schneider R, Gullo M, et al. Nogo-A antibody improves regeneration and locomotion of spinal cord-injured rats. Ann Neurol. 2005; 58:706–719. PMID: 16173073.
Article
22. Mandalari G, Genovese T, Bisignano C, Mazzon E, Wickham MS, Di Paola R, et al. Neuroprotective effects of almond skins in experimental spinal cord injury. Clin Nutr. 2011; 30:221–233. PMID: 20864228.
Article
23. Mao L, Wang H, Qiao L, Wang X. Disruption of Nrf2 enhances the upregulation of nuclear factor-kappaB activity, tumor necrosis factor-α, and matrix metalloproteinase-9 after spinal cord injury in mice. Mediators Inflamm. 2010; 2010:238321. PMID: 20862369.
24. Pahl HL. Activators and target genes of Rel/NF-kappaB transcription factors. Oncogene. 1999; 18:6853–6866. PMID: 10602461.
Article
25. Parr AM, Tator CH, Keating A. Bone marrow-derived mesenchymal stromal cells for the repair of central nervous system injury. Bone Marrow Transplant. 2007; 40:609–619. PMID: 17603514.
Article
26. Paterniti I, Genovese T, Crisafulli C, Mazzon E, Di Paola R, Galuppo M, et al. Treatment with green tea extract attenuates secondary inflammatory response in an experimental model of spinal cord trauma. Naunyn Schmiedebergs Arch Pharmacol. 2009; 380:179–192. PMID: 19337722.
Article
27. Pizzi M, Spano P. Distinct roles of diverse nuclear factor-kappaB complexes in neuropathological mechanisms. Eur J Pharmacol. 2006; 545:22–28. PMID: 16854410.
Article
28. Pollock G, Pennypacker KR, Mémet S, Israël A, Saporta S. Activation of NF-kappaB in the mouse spinal cord following sciatic nerve transection. Exp Brain Res. 2005; 165:470–477. PMID: 15912368.
Article
29. Rafati DS, Geissler K, Johnson K, Unabia G, Hulsebosch C, Nesic-Taylor O, et al. Nuclear factor-kappaB decoy amelioration of spinal cord injury-induced inflammation and behavior outcomes. J Neurosci Res. 2008; 86:566–580. PMID: 17918744.
Article
30. Schwaninger M, Inta I, Herrmann O. NF-kappaB signalling in cerebral ischaemia . Biochem Soc Trans. 2006; 34(Pt 6):1291–1294. PMID: 17073804.
31. Sekhon LH, Fehlings MG. Epidemiology, demographics, and pathophysiology of acute spinal cord injury. Spine (Phila Pa 1976). 2001; 26(24 Suppl):S2–S12. PMID: 11805601.
Article
32. Sribnick EA, Wingrave JM, Matzelle DD, Wilford GG, Ray SK, Banik NL. Estrogen attenuated markers of inflammation and decreased lesion volume in acute spinal cord injury in rats. J Neurosci Res. 2005; 82:283–293. PMID: 16130149.
Article
33. Tchivileva IE, Nackley AG, Qian L, Wentworth S, Conrad M, Diatchenko LB. Characterization of NF-kB-mediated inhibition of catechol-O-methyltransferase. Mol Pain. 2009; 5:13. PMID: 19291302.
Article
34. The Ministry of Science and Technology of the People's Republic of China. Guidance Suggestions for the Care and Use of Laboratory Animals. The People's Republic of China: The Ministry of Science and Technology of the People's Republic of China;2006.
35. Verma IM. Nuclear factor (NF)-kappaB proteins : therapeutic targets. Ann Rheum Dis. 2004; 63(Suppl 2):ii57–ii61. PMID: 15479873.
36. Willerth SM, Sakiyama-Elbert SE. Cell therapy for spinal cord regeneration. Adv Drug Deliv Rev. 2008; 60:263–276. PMID: 18029050.
Article
37. Wright KT, El Masri W, Osman A, Chowdhury J, Johnson WE. Concise review : bone marrow for the treatment of spinal cord injury : mechanisms and clinical applications. Stem Cells. 2011; 29:169–178. PMID: 21732476.
Article
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