SpringerLink
Log in

Inhibition of c-Jun N-terminal kinase prevents blood–brain barrier disruption and normalizes the expression of tight junction proteins clautin-5 and ZO-1 in a rat model of subarachnoid hemorrhage

  • Experimental research
  • Published:
Acta Neurochirurgica Aims and scope Submit manuscript

Abstract

Background

The c-Jun N-terminal kinase (JNK) proteins are encoded by three genes (JNK1, JNK2, and JNK3), giving rise to multiple isoforms via alternative splicing. JNK inhibition using a chemical inhibitor SP600125 confers neuroprotection in an animal model of subarachnoid hemorrhage (SAH). The aim of this study is to investigate whether the protective effects of SP600125 were associated with modulation of tight junction proteins including claudin-5 and ZO-1 and to define which JNK isoforms were involved in the early brain injury after SAH.

Methods

Seventy-five male Sprague Dawley rats (weighing 300–350 g) were randomly assigned to five groups (n = 15): (1) sham, (2) SAH, (3) SAH + DMSO (dimethyl sulfoxide), (4) SAH + 10 mg/kg SP600125, and (5) SAH + 30 mg/kg SP600125. SP600125 or DMSO was injected intraperitoneally 1 h before and 6 h after the induction of SAH. Animals from all the groups were killed 24 h after SAH, and brain tissues were dissected and subjected to electron microscopic examination, Western blot analysis, and histological evaluation.

Results

SP600125 pretreatment restored tight junctions and attenuated blood–brain barrier (BBB) disruption and cerebral edema after SAH, coupled with reduced apoptosis in the cerebral cortex. SP600125 exposure restored the reduced expression of both claudin-5 and ZO-1 following SAH and normalized the levels of JNK1 and JNK3.

Conclusion

Our data demonstrate that the JNK signaling plays an important role in the regulation of tight junction proteins and BBB integrity, and thus represents a promising target against brain injuries after SAH.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price includes VAT (France)

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Abbott NJ, Patabendige AA, Dolman DE, Yusof SR, Begley DJ (2010) Structure and function of the blood–brain barrier. Neurobiol Dis 37:13–25

    Article  PubMed  CAS  Google Scholar 

  2. Aveleira CA, Lin CM, Abcouwer SF, Ambrósio AF, Antonetti DA (2010) TNF-α signals through PKCζ/NF-κB to alter the tight junction complex and increase retinal endothelial cell permeability. Diabetes 59:2872–2882

    Article  PubMed  CAS  Google Scholar 

  3. Barnat M, Enslen H, Propst F, Davis RJ, Soares S, Nothias F (2010) Distinct roles of c-Jun N-terminal kinase isoforms in neurite initiation and elongation during axonal regeneration. J Neurosci 30:7804–7816

    Article  PubMed  CAS  Google Scholar 

  4. Bazzoni G (2006) Endothelial tight junctions: permeable barriers of the vessel wall. Thromb Haemost 95:36–42

    PubMed  CAS  Google Scholar 

  5. Bederson JB, Germano IM, Guarino L (1995) Cortical blood flow and cerebral perfusion pressure in a new noncraniotomy model of subarachnoid hemorrhage in the rat. Stroke 26:1086–1091

    Article  PubMed  CAS  Google Scholar 

  6. Broderick JP, Brott TG, Duldner JE, Tomsick T, Leach A (1994) Initial and recurrent bleeding are the major causes of death following subarachnoid hemorrhage. Stroke 25:1342–1347

    Article  PubMed  CAS  Google Scholar 

  7. Brown RC, Davis TP (2002) Calcium modulation of adherens and tight junction function: a potential mechanism for blood–brain barrier disruption after stroke. Stroke 33:1706–1711

    Article  PubMed  CAS  Google Scholar 

  8. Bogoyevitch MA (2006) The isoform-specific functions of the c-Jun N-terminal Kinases (JNKs): differences revealed by gene targeting. Bioessays 28:923–934

    Article  PubMed  CAS  Google Scholar 

  9. Cahill J, Calvert JW, Zhang JH (2006) Mechanisms of early brain injury after subarachnoid hemorrhage. J Cereb Blood Flow Metab 26:1341–1353

    Article  PubMed  CAS  Google Scholar 

  10. Crawford ED, Wells JA (2011) Caspase substrates and cellular remodeling. Annu Rev Biochem 80:1055–1087

    Article  PubMed  CAS  Google Scholar 

  11. Dhanasekaran DN, Reddy EP (2008) JNK signaling in apoptosis. Oncogene 27:6245–6251

    Article  PubMed  CAS  Google Scholar 

  12. Duris K, Manaenko A, Suzuki H, Rolland WB, Krafft PR, Zhang JH (2011) α7 Nicotinic acetylcholine receptor agonist PNU-282987 attenuates early brain injury in a perforation model of subarachnoid hemorrhage in rats. Stroke 42:3530–3536

    Google Scholar 

  13. Fanning AS, Jameson BJ, Jesaitis LA, Anderson JM (1998) The tight junction protein ZO-1 establishes a link between the transmembrane protein occludin and the actin cytoskeleton. J Biol Chem 273:29745–29753

    Article  PubMed  CAS  Google Scholar 

  14. Fontijn RD, Volger OL, Fledderus JO, Reijerkerk A, de Vries HE, Horrevoets AJ (2008) SOX-18 controls endothelial-specific claudin-5 gene expression and barrier function. Am J Physiol Heart Circ Physiol 294:H891–H900

    Article  PubMed  CAS  Google Scholar 

  15. Gupta S, Barrett T, Whitmarsh AJ, Cavanagh J, Sluss HK, Dérijard B, Davis RJ (1996) Selective interaction of JNK protein kinase isoforms with transcription factors. EMBO J 15:2760–2770

    PubMed  CAS  Google Scholar 

  16. Hishikawa T, Ono S, Ogawa T, Tokunaga K, Sugiu K, Date I (2008) Effects of deferoxamine-activated hypoxia-inducible factor-1 on the brainstem after subarachnoid hemorrhage in rats. Neurosurgery 62:232–240

    Article  PubMed  Google Scholar 

  17. Kuan CY, Yang DD, Samanta Roy DR, Davis RJ, Rakic P, Flavell RA (1999) The Jnk1 and Jnk2 protein kinases are required for regional specific apoptosis during early brain development. Neuron 22:667–676

    Article  PubMed  CAS  Google Scholar 

  18. Kusaka G, Ishikawa M, Nanda A, Granger DN, Zhang JH (2004) Signaling pathways for early brain injury after subarachnoid hemorrhage. J Cereb Blood Flow Metab 24:916–925

    Article  PubMed  CAS  Google Scholar 

  19. László FA, Varga C, Dóczi T (1995) Cerebral oedema after subarachnoid haemorrhage. Pathogenetic significance of vasopressin. Acta Neurochir (Wien) 133:122–133

    Article  Google Scholar 

  20. Liu J, Lin A (2005) Role of JNK activation in apoptosis: a double-edged sword. Cell Res 15:36–42

    Article  PubMed  Google Scholar 

  21. Li Y, Tang J, Khatibi NH, Zhu M, Chen D, Zheng W, Wang S (2010) Ginsenoside Rbeta1 reduces neurologic damage, is anti-apoptotic, and down-regulates p53 and BAX in subarachnoid hemorrhage. Curr Neurovasc Res 7:85–94

    Article  PubMed  Google Scholar 

  22. Matz PG, Fujimura M, Lewen A, Morita-Fujimura Y, Chan PH (2001) Increased cytochrome c-mediated DNA fragmentation and cell death in manganesesuperoxide dismutase-deficient mice after exposure to subarachnoid hemolysate. Stroke 32:506–515

    Article  PubMed  CAS  Google Scholar 

  23. Mikawa S, Kinouchi H, Kamii H, Gobbel GT, Chen SF, Carlson E, Epstein CJ, Chan PH (1996) Attenuation of acute and chronic damage following traumatic brain injury in copper, zinc-superoxide dismutase transgenic mice. J Neurosurg 85:885–891

    Article  PubMed  CAS  Google Scholar 

  24. Morita K, Sasaki H, Furuse M, Tsukita S (1999) Endothelial claudin: claudin-5/TMVCF constitutes tight junction strands in endothelial cells. J Cell Biol 147:185–194

    Article  PubMed  CAS  Google Scholar 

  25. Nag S, Kapadia A, Stewart DJ (2011) Review: molecular pathogenesis of blood–brain barrier breakdown in acute brain injury. Neuropathol Appl Neurobiol 37:3–23

    Article  PubMed  CAS  Google Scholar 

  26. Nitta T, Hata M, Gotoh S, Seo Y, Sasaki H, Hashimoto N, Furuse M, Tsukita S (2003) Size-selective loosening of the blood–brain barrier in claudin-5-deficient mice. J Cell Biol 161:653–660

    Article  PubMed  CAS  Google Scholar 

  27. Ostrowski RP, Colohan AR, Zhang JH (2005) Mechanisms of hyperbaric oxygen-induced neuroprotection in a rat model of subarachnoid hemorrhage. J Cereb Blood Flow Metab 25:554–571

    Article  PubMed  CAS  Google Scholar 

  28. Ostrowski RP, Colohan AR, Zhang JH (2006) Molecular mechanisms of early brain injury after subarachnoid hemorrhage. Neurol Res 28:399–414

    Article  PubMed  CAS  Google Scholar 

  29. Park JE, Lee JA, Park SG, Lee DH, Kim SJ, Kim HJ, Uchida C, Uchida T, Park BC, Cho S (2012) A critical step for JNK activation: isomerization by the prolyl isomerase Pin1. Cell Death Differ 19:153–161

    Google Scholar 

  30. Porter AG, Jänicke RU (1999) Emerging roles of caspase-3 in apoptosis. Cell Death Differ 6:99–104

    Article  PubMed  CAS  Google Scholar 

  31. Schievink WI, Riedinger M, Jhutty TK, Simon P (2004) Racial disparities in subarachnoid hemorrhage mortality: Los Angeles County, California, 1985–1998. Neuroepidemiology 23:299–305

    Article  PubMed  Google Scholar 

  32. Singh R, Wang Y, **ang Y, Tanaka KE, Gaarde WA, Czaja MJ (2009) Differential effects of JNK1 and JNK2 inhibition on murine steatohepatitis and insulin resistance. Hepatology 49:87–96

    Article  PubMed  CAS  Google Scholar 

  33. Suzuki H, Hasegawa Y, Kanamaru K, Zhang JH (2011) Mitogen-activated protein kinases in cerebral vasospasm after subarachnoid hemorrhage: a review. Acta Neurochir Suppl 110:133–139

    Article  PubMed  Google Scholar 

  34. Yan J, Li L, Khatibi NH, Yang L, Wang K, Zhang W, Martin RD, Han J, Zhang J, Zhou C (2011) Blood–brain barrier disruption following subarchnoid hemorrhage may be faciliated through PUMA induction of endothelial cell apoptosis from the endoplasmic reticulum. Exp Neurol 230:240–247

    Article  PubMed  CAS  Google Scholar 

  35. Yatsushige H, Ostrowski RP, Tsubokawa T, Colohan A, Zhang JH (2007) Role of c-Jun N-terminal kinase in early brain injury after subarachnoid hemorrhage. J Neurosci Res 85:1436–1448

    Article  PubMed  CAS  Google Scholar 

  36. Yatsushige H, Yamaguchi-Okada M, Zhou C, Calvert JW, Cahill J, Colohan AR, Zhang JH (2008) Inhibition of c-Jun N-terminal kinase pathway attenuates cerebral vasospasm after experimental subarachnoid hemorrhage through the suppression of apoptosis. Acta Neurochir Suppl 104:27–31

    Article  PubMed  CAS  Google Scholar 

  37. Zhou Y, Martin RD, Zhang JH (2011) Advances in experimental subarachnoid hemorrhage. Acta Neurochir Suppl 110:15–21

    Article  PubMed  Google Scholar 

Download references

Conflicts of interest

None.

Author information

Authors and Affiliations

  1. Department of Neurosurgery, Sheng**g Hospital of China Medical University, Shenyang, 110004, China

    Duo Chen, **ang-tai Wei, Jun-hong Guan, Jiang-wei Yuan, Yu-tao Peng, Lei Song & Yun-hui Liu

Authors
  1. Duo Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. **ang-tai Wei
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jun-hong Guan
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jiang-wei Yuan
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yu-tao Peng
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Lei Song
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yun-hui Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jun-hong Guan.

Additional information

Comment

The authors are investigating the mechanism underlying their previous observation that inhibition of JNK activation with SP600125 is neuroprotective following experimental rat SAH. The hypothesis is that administration of this agent preserves tight junctions at the capillary endothelium level and prevents BBB disruption following SAH.

The investigators’ approach is clever and multifaceted, looking at morphologic brain endpoints (edema, BBB disruption) and mechanistic endpoints (ultrastructure of the BBB, protein structure and regulation at the BBB). Consideration of the method shows that techniques are standard and the experimental observations are appropriately controlled. The SAH was created with a sharpened suture endovascular carotid perforation, and agent was administered pre- and post-injury. BBB permeability was evaluated with Evans blue. Selected protein analysis was with Western blot and apoptosis was measured with TUNEL counts. Cellular ultrastructure was assessed with TEM.

Experimental observations are both clinical and mechanistic. Clinically, SP600125 administration decreased Evans blue dye-observed BBB disruption. Brain water content (reflecting edema) was likewise statistically diminished in the treated group, and apoptosis was decreased as well. Mechanistically, by TEM there was more tight junction integrity in the treatment group. Integrity proteins at the BBB claudin-5 and ZO-1 were preserved by SP600125 dosing, while JNK1 and JNK3 were decreased. The implication is that JNK signals the deleterious BBB changes following experimental SAH.

As the authors conclude, targeting JNK signalling may be one way to increase neuroprotection and neural preservation following SAH. Certainly there are many inflection points in the SAH injury process. This excellent manuscript, by experienced and sophisticated investigators, rigorously follows one such thread, and as such adds value to our knowledge of SAH brain injury, although any clinical application, if appropriate, remains far away.

Christopher M. Loftus

Philadelphia, PA

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary Fig. S1

Determination of mortality rates 24 h after subarachnoid hemorrhage (SAH). SP(10), SP(30) treatment with 10 and 30 mg/kg SP600125, respectively. n.s. no significance (DOC 44 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chen, D., Wei, Xt., Guan, Jh. et al. Inhibition of c-Jun N-terminal kinase prevents blood–brain barrier disruption and normalizes the expression of tight junction proteins clautin-5 and ZO-1 in a rat model of subarachnoid hemorrhage. Acta Neurochir 154, 1469–1476 (2012). https://doi.org/10.1007/s00701-012-1328-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00701-012-1328-y

Keywords

Search

Navigation