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Ursodeoxycholic Acid Inhibits Inflammatory Responses and Promotes Functional Recovery After Spinal Cord Injury in Rats

Abstract

The aim of this study was to investigate the anti-inflammatory effects by ursodeoxycholic acid (UDCA) in rats with a spinal cord injury (SCI). A moderate mechanical compression injury was imposed on adult Sprague-Dawley (SD) rats. The post-injury locomotor functions were assessed using the Basso, Beattie, and Bresnahan (BBB) locomotor scale and the tissue volume of the injured region was analyzed using hematoxylin and eosin staining. The pro-inflammatory factors were evaluated by immunofluorescence (IF) staining, a quantitative real-time polymerase chain reaction (qRT-PCR), and enzyme-linked immunosorbent assay (ELISA). The phosphorylation of the extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 in mitogen-activated protein kinase (MAPK) signaling pathways related to inflammatory responses were measured by Western blot assays. UDCA improved the BBB scores and promoted the recovery of the spinal cord lesions. UDCA inhibited the expression of glial fibrillary acidic protein (GFAP), tumor necrosis factor-α (TNF-α), ionized calcium-binding adapter molecule 1 (iba1), and inducible nitric oxide synthase (iNOS). UDCA decreased the pro-inflammatory cytokines of TNF-α, interleukin 1-β (IL-1β), and interleukin 6 (IL-6) in the mRNA and protein levels. UDCA increased the anti-inflammatory cytokine interleukin 10 (IL-10) in the mRNA and protein levels. UDCA suppressed the phosphorylation of ERK, JNK, and the p38 signals. UDCA reduces pro-inflammatory responses and promotes functional recovery in SCI in rats. These results suggest that UDCA is a potential therapeutic drug for SCI.

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References

  1. Thuret S, Moon LD, Gage FH (2006) Therapeutic interventions after spinal cord injury. Nat Rev Neurosci 7(8):628–643

    CAS  Article  PubMed  Google Scholar 

  2. Xu J, Kim GM, Ahmed SH, Xu J, Yan P, Xu XM, Hsu CY (2001) Glucocorticoid receptor-mediated suppression of activator protein-1 activation and matrix metalloproteinase expression after spinal cord injury. J Neurosci 21(1):92–97

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  3. Noble LJ, Donovan F, Igarashi T, Goussev S, Werb Z (2002) Matrix metalloproteinases limit functional recovery after spinal cord injury by modulation of early vascular events. J Neurosci 22(17):7526–7535

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  4. Hausmann ON (2003) Post-traumatic inflammation following spinal cord injury. Spinal Cord 41(7):369–378

    CAS  Article  PubMed  Google Scholar 

  5. Qiao F, Atkinson C, Kindy MS, Shunmugavel A, Morgan BP, Song H, Tomlinson S (2010) The alternative and terminal pathways of complement mediate post-traumatic spinal cord inflammation and injury. Am J Pathol 177(6):3061–3070

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  6. Beck KD, Nguyen HX, Galvan MD, Salazar DL, Woodruff TM, Anderson AJ (2010) Quantitative analysis of cellular inflammation after traumatic spinal cord injury: evidence for a multiphasic inflammatory response in the acute to chronic environment. Brain 133(Pt 2):433–447

    Article  PubMed  PubMed Central  Google Scholar 

  7. Beattie MS (2004) Inflammation and apoptosis: linked therapeutic targets in spinal cord injury. Trends Mol Med 10(12):580–583

    CAS  Article  PubMed  Google Scholar 

  8. Bartholdi D, Schwab ME (1995) Methylprednisolone inhibits early inflammatory processes but not ischemic cell death after experimental spinal cord lesion in the rat. Brain Res 672(1–2):177–186

    CAS  Article  PubMed  Google Scholar 

  9. Gerndt SJ, Rodriguez JL, Pawlik JW, Taheri PA, Wahl WL, Micheals AJ, Papadopoulos SM (1997) Consequences of high-dose steroid therapy for acute spinal cord injury. J Trauma 42(2):279–284

    CAS  Article  PubMed  Google Scholar 

  10. Qian T, Guo X, Levi AD, Vanni S, Shebert RT, Sipski ML (2005) High-dose methylprednisolone may cause myopathy in acute spinal cord injury patients. Spinal Cord 43(4):199–203

    CAS  Article  PubMed  Google Scholar 

  11. Shi KQ, Fan YC, Liu WY, Li LF, Chen YP, Zheng MH (2012) Traditional Chinese medicines benefit to nonalcoholic fatty liver disease: a systematic review and meta-analysis. Mol Biol Rep 39(10):9715–9722

    CAS  Article  PubMed  Google Scholar 

  12. Bachrach WH, Hofmann AF (1982) Ursodeoxycholic acid in the treatment of cholesterol cholelithiasis. part I. Dig Dis Sci 27(8):737–761

    CAS  Article  PubMed  Google Scholar 

  13. Beuers U, Boyer JL, Paumgartner G (1998) Ursodeoxycholic acid in cholestasis: potential mechanisms of action and therapeutic applications. Hepatology 28(6):1449–1453

    CAS  Article  PubMed  Google Scholar 

  14. Cirillo NW, Zwas FR (1994) Ursodeoxycholic acid in the treatment of chronic liver disease. Am J Gastroenterol 89(9):1447–1452

    CAS  PubMed  Google Scholar 

  15. Lazaridis KN, Gores GJ, Lindor KD (2001) Ursodeoxycholic acid mechanisms of action and clinical use in hepatobiliary disorders. J Hepatol 35(1):134–146

    CAS  Article  PubMed  Google Scholar 

  16. Ko WK, Lee SH, Kim SJ, Jo MJ, Kumar H, Han IB, Sohn S (2017) Anti-inflammatory effects of ursodeoxycholic acid by lipopolysaccharide-stimulated inflammatory responses in RAW 264.7 macrophages. PLoS One 12(6):e0180673

    Article  PubMed  PubMed Central  Google Scholar 

  17. Fleming JC, Norenberg MD, Ramsay DA, Dekaban GA, Marcillo AE, Saenz AD, Pasquale-Styles M, Dietrich WD et al (2006) The cellular inflammatory response in human spinal cords after injury. Brain 129(Pt 12):3249–3269

    Article  PubMed  Google Scholar 

  18. Donnelly DJ, Popovich PG (2008) Inflammation and its role in neuroprotection, axonal regeneration and functional recovery after spinal cord injury. Exp Neurol 209(2):378–388

    CAS  Article  PubMed  Google Scholar 

  19. Ropper AE, Zeng X, Anderson JE, Yu D, Han I, Haragopal H, Teng YD (2015) An efficient device to experimentally model compression injury of mammalian spinal cord. Exp Neurol 271:515–523

    Article  PubMed  Google Scholar 

  20. Basso DM, Beattie MS, Bresnahan JC (1995) A sensitive and reliable locomotor rating scale for open field testing in rats. J Neurotrauma 12(1):1–21

    CAS  Article  PubMed  Google Scholar 

  21. Kumar H, Jo MJ, Choi H, Muttigi MS, Shon S, Kim BJ, Lee SH, Han IB (2017) Matrix Metalloproteinase-8 inhibition prevents disruption of blood-spinal cord barrier and attenuates inflammation in rat model of spinal cord injury. Mol Neurobiol 54:3578–3590

    CAS  Article  PubMed  Google Scholar 

  22. Collins TJ (2007) ImageJ for microscopy. BioTechniques 43(1 Suppl):25–30

    Article  PubMed  Google Scholar 

  23. Schmittgen TD, Livak KJ (2008) Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc 3(6):1101–1108

    CAS  Article  PubMed  Google Scholar 

  24. Hayashi M, Ueyama T, Nemoto K, Tamaki T, Senba E (2000) Sequential mRNA expression for immediate early genes, cytokines, and neurotrophins in spinal cord injury. J Neurotrauma 17(3):203–218

    CAS  Article  PubMed  Google Scholar 

  25. Saiwai H, Kumamaru H, Ohkawa Y, Kubota K, Kobayakawa K, Yamada H, Yokomizo T, Iwamoto Y et al (2013) Ly6C+ Ly6G-myeloid-derived suppressor cells play a critical role in the resolution of acute inflammation and the subsequent tissue repair process after spinal cord injury. J Neurochem 125(1):74–88

    CAS  Article  PubMed  Google Scholar 

  26. Bethea JR, Nagashima H, Acosta MC, Briceno C, Gomez F, Marcillo AE, Loor K, Green J et al (1999) Systemically administered interleukin-10 reduces tumor necrosis factor-alpha production and significantly improves functional recovery following traumatic spinal cord injury in rats. J Neurotrauma 16(10):851–863

    CAS  Article  PubMed  Google Scholar 

  27. Sweitzer SM, Colburn RW, Rutkowski M, DeLeo JA (1999) Acute peripheral inflammation induces moderate glial activation and spinal IL-1beta expression that correlates with pain behavior in the rat. Brain Res 829(1–2):209–221

    CAS  Article  PubMed  Google Scholar 

  28. Yin X, Yin Y, Cao FL, Chen YF, Peng Y, Hou WG, Sun SK, Luo ZJ (2012) Tanshinone IIA attenuates the inflammatory response and apoptosis after traumatic injury of the spinal cord in adult rats. PLoS One 7(6):e38381

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  29. Bareyre FM, Schwab ME (2003) Inflammation, degeneration and regeneration in the injured spinal cord: Insights from DNA microarrays. Trends Neurosci 26(10):555–563

    CAS  Article  PubMed  Google Scholar 

  30. Genovese T, Esposito E, Mazzon E, Muia C, Di Paola R, Meli R, Bramanti P, Cuzzocrea S (2008) Evidence for the role of mitogen-activated protein kinase signaling pathways in the development of spinal cord injury. J Pharmacol Exp Ther 325(1):100–114

    CAS  Article  PubMed  Google Scholar 

  31. Hiyoshi T, Kambe D, Karasawa J, Chaki S (2014) Differential effects of NMDA receptor antagonists at lower and higher doses on basal gamma band oscillation power in rat cortical electroencephalograms. Neuropharmacology 85:384–396

    CAS  Article  PubMed  Google Scholar 

  32. Cheng Y, Tauschel HD, Nilsson A, Duan RD (1999) Ursodeoxycholic acid increases the activities of alkaline sphingomyelinase and caspase-3 in the rat colon. Scand J Gastroenterol 34(9):915–920

    CAS  Article  PubMed  Google Scholar 

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Funding

This work was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT, and future Planning (NRF-2016M3A9E8941668).

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Correspondence to In-Bo Han or Seil Sohn.

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The authors declare that they have no conflict of interest.

Electronic Supplementary Material

Fig. S1
figure 9

Quantification of GFAP and TNF-α in injured spinal cords. (A) Quantitative analysis of the fluorescence intensity for GFAP. (B) Analysis of the quantitative fluorescence intensity for TNF-α. Results are the mean ± SEM: *p < 0.05 and **p < 0.01, significant differences among three groups were demonstrated. (GIF 18 kb)

Fig. S2
figure 10

Quantification of iba1 and iNOS in injured spinal cords. (A) Quantitative analysis of the fluorescence intensity for iba1. (B) Analysis of the quantitative fluorescence intensity for iNOS. Results are the mean ± SEM: *p < 0.05 and **p < 0.01, significant differences among three groups were demonstrated. (GIF 15 kb)

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Ko, WK., Kim, S.J., Jo, MJ. et al. Ursodeoxycholic Acid Inhibits Inflammatory Responses and Promotes Functional Recovery After Spinal Cord Injury in Rats. Mol Neurobiol 56, 267–277 (2019). https://doi.org/10.1007/s12035-018-0994-z

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  • DOI: https://doi.org/10.1007/s12035-018-0994-z

Keywords

  • Ursodeoxycholic acid
  • Spinal cord injury
  • BBB score
  • MAPK signaling
  • TNF-α
  • Anti-inflammation