J Korean Med Sci.  2015 Jul;30(7):943-952. 10.3346/jkms.2015.30.7.943.

Agmatine Attenuates Brain Edema and Apoptotic Cell Death after Traumatic Brain Injury

• Kim JY
• Lee YW
• Kim JH
• Lee WT
• Park KA
• Lee JE

• KMID: 2164481
• DOI: http://doi.org/10.3346/jkms.2015.30.7.943

Abstract

Traumatic brain injury (TBI) is associated with poor neurological outcome, including necrosis and brain edema. In this study, we investigated whether agmatine treatment reduces edema and apoptotic cell death after TBI. TBI was produced by cold injury to the cerebral primary motor cortex of rats. Agmatine was administered 30 min after injury and once daily until the end of the experiment. Animals were sacrificed for analysis at 1, 2, or 7 days after the injury. Various neurological analyses were performed to investigate disruption of the blood-brain barrier (BBB) and neurological dysfunction after TBI. To examine the extent of brain edema after TBI, the expression of aquaporins (AQPs), phosphorylation of mitogen-activated protein kinases (MAPKs), and nuclear translocation of nuclear factor-kappaB (NF-kappaB) were investigated. Our findings demonstrated that agmatine treatment significantly reduces brain edema after TBI by suppressing the expression of AQP1, 4, and 9. In addition, agmatine treatment significantly reduced apoptotic cell death by suppressing the phosphorylation of MAPKs and by increasing the nuclear translocation of NF-kappaB after TBI. These results suggest that agmatine treatment may have therapeutic potential for brain edema and neural cell death in various central nervous system diseases.

Keyword

Agmatine; Traumatic Brain Injury; Brain Edema; Blood-Brain Barrier; Aquaporins; Mitogen-Activated Protein Kinases

MeSH Terms

Active Transport, Cell Nucleus/drug effects
Agmatine/*therapeutic use
Animals
Apoptosis/*drug effects
Aquaporins/metabolism
Blood-Brain Barrier/physiopathology
Brain Edema/*drug therapy
Brain Injuries/*pathology
Male
Mitogen-Activated Protein Kinases/metabolism
Motor Cortex/*pathology
NF-kappa B/metabolism
Phosphorylation/drug effects
Rats
Rats, Sprague-Dawley
Agmatine
Aquaporins
NF-kappa B
Mitogen-Activated Protein Kinases

Figure

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  Fig. 1 Dorsal and coronal views of the rat skull and experimental scheme. Experimental animals were anesthetized and placed on the stereotactic frame. Craniotomy was performed above the right forelimb motor cortex at +2 mm anterior and -2 mm posterior from Bregma and +1-5 mm lateral from the midline. The representative figure is from "The Rat Brain in Stereotaxic Coordinates," fourth edition, George Paxinos & Charles Watson, Academic Press 1998.

• 

  Fig. 2 Agmatine treatment significantly attenuated brain edema and neural cell death after TBI. Measurements were made of the Evans Blue extravasations (A), the Evans Blue stained area (mm2) (B), swelling volume (C), and brain water content (D). Hematoxylin and eosin (H & E) staining (E), relative necrotic area measurement (F), and TUNEL-positive cells in the ipsilateral cortex (G) were all significantly suppressed. Data are expressed as the mean±SEM (*P < 0.05 vs. EC). EC (n=4), experimental control group; Agm (n=4), agmatine treatment group.

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  Fig. 3 Agmatine ameliorated neurological behavior dysfunction after TBI. The rotarod test (A) and limb placement test (B) were conducted in control and experimental animals. Agmatine administration promoted motor function under both conditions. Data are expressed as the mean±SEM (*P < 0.05 vs. EC).

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  Fig. 4 Agmatine attenuated edema formation by suppressing AQPs after TBI. Agmatine attenuated aquaporin expression 7 days after TBI. Immunohistochemistry of AQP1, 4, and 9 (A-C) and the protein expression of AQP1, 4, and 9 (D) were analyzed. Data are expressed as the mean±SEM (*P < 0.05 vs. EC).

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  Fig. 5 Agmatine suppressed JNK and P38 MAPK phosphorylation after TBI. Representative expression of phosphorylated ERK (A), JNK (B), and P38 MAPK (C) in control animals are shown in immunohistochemistry (A-C) and immune blotting (D). Agmatine suppressed the phosphorylation of JNK (F) and P38 MAPK (G) 1 day after TBI. However, agmatine treatment increased the phosphorylation of ERK (E) after TBI. Data are expressed as the mean±SEM (*P < 0.05 vs. EC).

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  Fig. 6 Agmatine promoted NF-κB p65 translocation into the nucleus after TBI. Protein expression of cytosolic NF-κB p65 (A) and nuclear NF-κB p65 (B) are shown. Agmatine caused significant translocation of NF-κB p65 into the nucleus 1 day after TBI. Optical density of cytosolic fraction (C) and nuclear fraction (D) were analyzed. Data are expressed as the mean±SEM (*P < 0.05 vs. EC).

Reference

1. Reilly PL. Brain injury: the pathophysiology of the first hours.'Talk and Die revisited'. J Clin Neurosci. 2001; 8:398–403.

2. McIntosh TK, Juhler M, Wieloch T. Novel pharmacologic strategies in the treatment of experimental traumatic brain injury: 1998. J Neurotrauma. 1998; 15:731–769.

3. Murakami K, Kondo T, Yang G, Chen SF, Morita-Fujimura Y, Chan PH. Cold injury in mice: a model to study mechanisms of brain edema and neuronal apoptosis. Prog Neurobiol. 1999; 57:289–299.

4. Hu BR, Liu CL, Park DJ. Alteration of MAP kinase pathways after transient forebrain ischemia. J Cereb Blood Flow Metab. 2000; 20:1089–1095.

5. Sugino T, Nozaki K, Hashimoto N. Activation of mitogen-activated protein kinases in gerbil hippocampus with ischemic tolerance induced by 3-nitropropionic acid. Neurosci Lett. 2000; 278:101–104.

6. Mori T, Wang X, Jung JC, Sumii T, Singhal AB, Fini ME, Dixon CE, Alessandrini A, Lo EH. Mitogen-activated protein kinase inhibition in traumatic brain injury: in vitro and in vivo effects. J Cereb Blood Flow Metab. 2002; 22:444–452.

7. Otani N, Nawashiro H, Fukui S, Nomura N, Yano A, Miyazawa T, Shima K. Differential activation of mitogen-activated protein kinase pathways after traumatic brain injury in the rat hippocampus. J Cereb Blood Flow Metab. 2002; 22:327–334.

8. Arima H, Yamamoto N, Sobue K, Umenishi F, Tada T, Katsuya H, Asai K. Hyperosmolar mannitol simulates expression of aquaporins 4 and 9 through a p38 mitogen-activated protein kinase-dependent pathway in rat astrocytes. J Biol Chem. 2003; 278:44525–44534.

9. Nito C, Kamada H, Endo H, Narasimhan P, Lee YS, Chan PH. Involvement of mitogen-activated protein kinase pathways in expression of the water channel protein aquaporin-4 after ischemia in rat cortical astrocytes. J Neurotrauma. 2012; 29:2404–2412.

10. Mattson MP, Culmsee C, Yu Z, Camandola S. Roles of nuclear factor kappaB in neuronal survival and plasticity. J Neurochem. 2000; 74:443–456.

11. Karin M, Lin A. NF-kappaB at the crossroads of life and death. Nat Immunol. 2002; 3:221–227.

12. Li G, Regunathan S, Barrow CJ, Eshraghi J, Cooper R, Reis DJ. Agmatine: an endogenous clonidine-displacing substance in the brain. Science. 1994; 263:966–969.

13. Piletz JE, Chikkala DN, Ernsberger P. Comparison of the properties of agmatine and endogenous clonidine-displacing substance at imidazoline and alpha-2 adrenergic receptors. J Pharmacol Exp Ther. 1995; 272:581–587.

14. Olmos G, DeGregorio-Rocasolano N, Paz Regalado M, Gasull T, Assumpció Boronat M, Trullas R, Villarroel A, Lerma J, García-Sevilla JA. Protection by imidazol(ine) drugs and agmatine of glutamate-induced neurotoxicity in cultured cerebellar granule cells through blockade of NMDA receptor. Br J Pharmacol. 1999; 127:1317–1326.

15. Halaris A, Plietz J. Agmatine : metabolic pathway and spectrum of activity in brain. CNS Drugs. 2007; 21:885–900.

16. Feng Y, Piletz JE, Leblanc MH. Agmatine suppresses nitric oxide production and attenuates hypoxic-ischemic brain injury in neonatal rats. Pediatr Res. 2002; 52:606–611.

17. Gilad GM, Gilad VH. Accelerated functional recovery and neuroprotection by agmatine after spinal cord ischemia in rats. Neurosci Lett. 2000; 296:97–100.

18. Kim JH, Yenari MA, Giffard RG, Cho SW, Park KA, Lee JE. Agmatine reduces infarct area in a mouse model of transient focal cerebral ischemia and protects cultured neurons from ischemia-like injury. Exp Neurol. 2004; 189:122–130.

19. Yang XC, Reis DJ. Agmatine selectively blocks the N-methyl-D-aspartate subclass of glutamate receptor channels in rat hippocampal neurons. J Pharmacol Exp Ther. 1999; 288:544–549.

20. Gilad GM, Salame K, Rabey JM, Gilad VH. Agmatine treatment is neuroprotective in rodent brain injury models. Life Sci. 1996; 58:PL 41–PL 46.

21. Lee WT, Hong S, Yoon SH, Kim JH, Park KA, Seong GJ, Lee JE. Neuroprotective effects of agmatine on oxygen-glucose deprived primary-cultured astrocytes and nuclear translocation of nuclear factor-kappa B. Brain Res. 2009; 1281:64–70.

22. Kim JH, Lee YW, Park KA, Lee WT, Lee JE. Agmatine attenuates brain edema through reducing the expression of aquaporin-1 after cerebral ischemia. J Cereb Blood Flow Metab. 2010; 30:943–949.

23. Kuo JR, Lo CJ, Chio CC, Chang CP, Lin MT. Resuscitation from experimental traumatic brain injury by agmatine therapy. Resuscitation. 2007; 75:506–514.

24. Wang CC, Chio CC, Chang CH, Kuo JR, Chang CP. Beneficial effect of agmatine on brain apoptosis, astrogliosis, and edema after rat transient cerebral ischemia. BMC Pharmacol. 2010; 10:11.

25. Kuo JR, Lo CJ, Chang CP, Lin KC, Lin MT, Chio CC. Agmatine-promoted angiogenesis, neurogenesis, and inhibition of gliosis-reduced traumatic brain injury in rats. J Trauma. 2011; 71:E87–E93.

26. Nag S. Cold-injury of the cerebral cortex: immunolocalization of cellular proteins and blood-brain barrier permeability studies. J Neuropathol Exp Neurol. 1996; 55:880–888.

27. Gröger M, Lebesgue D, Pruneau D, Relton J, Kim SW, Nussberger J, Plesnila N. Release of bradykinin and expression of kinin B2 receptors in the brain: role for cell death and brain edema formation after focal cerebral ischemia in mice. J Cereb Blood Flow Metab. 2005; 25:978–989.

28. Puurunen K, Jolkkonen J, Sirvio J, Haapalinna A, Sivenius J. Selegiline combined with enriched-environment housing attenuates spatial learning deficits following focal cerebral ischemia in rats. Exp Neurol. 2001; 167:348–355.

29. Ahmed KM, Dong S, Fan M, Li JJ. Nuclear factor-kappaB p65 inhibits mitogen-activated protein kinase signaling pathway in radioresistant breast cancer cells. Mol Cancer Res. 2006; 4:945–955.

30. Impey S, Obrietan K, Storm DR. Making new connections: role of ERK/MAP kinase signaling in neuronal plasticity. Neuron. 1999; 23:11–14.

31. Arndt MA, Battaglia V, Parisi E, Lortie MJ, Isome M, Baskerville C, Pizzo DP, Ientile R, Colombatto S, Toninello A, et al. The arginine metabolite agmatine protects mitochondrial function and confers resistance to cellular apoptosis. Am J Physiol Cell Physiol. 2009; 296:C1411–C1419.

32. Condello S, Currò M, Ferlazzo N, Caccamo D, Satriano J, Ientile R. Agmatine effects on mitochondrial membrane potential and NF-kappaB activation protect against rotenone-induced cell damage in human neuronal-like SH-SY5Y cells. J Neurochem. 2011; 116:67–75.

33. de Vries HE, Blom-Roosemalen MC, van Oosten M, de Boer AG, van Berkel TJ, Breimer DD, Kuiper J. The influence of cytokines on the integrity of the blood-brain barrier in vitro. J Neuroimmunol. 1996; 64:37–43.

34. Morganti-Kossmann MC, Rancan M, Stahel PF, Kossmann T. Inflammatory response in acute traumatic brain injury: a double-edged sword. Curr Opin Crit Care. 2002; 8:101–105.

35. Liang D, Bhatta S, Gerzanich V, Simard JM. Cytotoxic edema: mechanisms of pathological cell swelling. Neurosurg Focus. 2007; 22:E2.

36. Guo Q, Sayeed I, Baronne LM, Hoffman SW, Guennoun R, Stein DG. Progesterone administration modulates AQP4 expression and edema after traumatic brain injury in male rats. Exp Neurol. 2006; 198:469–478.

37. Manley GT, Fujimura M, Ma T, Noshita N, Filiz F, Bollen AW, Chan P, Verkman AS. Aquaporin-4 deletion in mice reduces brain edema after acute water intoxication and ischemic stroke. Nat Med. 2000; 6:159–163.

38. Badaut J, Lasbennes F, Magistretti PJ, Regli L. Aquaporins in brain: distribution, physiology, and pathophysiology. J Cereb Blood Flow Metab. 2002; 22:367–378.

39. Papadopoulos MC, Krishna S, Verkman AS. Aquaporin water channels and brain edema. Mt Sinai J Med. 2002; 69:242–248.