Melatonin treatment reduces astrogliosis and apoptosis in rats with traumatic brain injury

Document Type: Original Article

Authors

1 Department of Anatomical Sciences, Afzalipour School of Medicine, Kerman University of Medical Sciences, Kerman, Iran

2 Neuroscience Research Center, Institute of Neuropharmacology, Kerman University of Medical Sciences, Kerman, Iran

3 Department of Physiology, Afzalipour School of Medicine, Kerman University of Medical Sciences, Kerman, Iran

4 Department of Pathology, Afzalipour School of Medicine, Kerman University of Medical Sciences, Kerman, Iran

Abstract

Objective(s):Melatonin is known as an anti-inflammatory agent, and it has been proven to exert neuroprotection through inhibition of cell death (apoptosis) in several models of brain injury.Secondary injury following the primary traumatic brain injury (TBI) results in glial cells activation, especially astrocytes. In fact, astrocyte activation causes the production of pro-inflammatory cytokines that may lead to secondary injury. Since most TBI research studies have focused on injured neurons and paid little attention to glial cells, the aim of current study was to investigate the effects of melatonin against astrocytes activation (astrogliosis), as well as inhibition of apoptosis in brain tissue of male rats after TBI.
Materials and Methods: The animals were randomly allocated into five groups: sham group, TBI+ vehicle group (1% ethanol in saline) and TBI+ melatonin groups (5 mg/kg, 10 mg/kg and 20 mg/kg). All rats were intubated and then exposed to diffuse TBI, except for the sham group. Immunohistochemical methods were conducted using glial fibrillary acidic protein (GFAP) marker and TUNEL assay to evaluate astrocyte reactivity and cell death, respectively.
Results: The results showed that based on the number of GFAP positive astrocytes in brain cortex, astrogliosis was reduced significantly (P<0.05) in melatonin- treated groups (no dose dependent) compared to the vehicle group. Furthermore, based on TUNEL results, melatonin treatment considerably reduced the number of apoptotic cells (P<0.05).
Conclusion:In total, the present findings suggest that melatonin treatment following TBI diminishes astrocyte reactivity and neuronal cells apoptosis in brain cortex in the rat model.

Keywords


1. Chua KSG NY, Yap SGM, Bok C. A brief review of traumatic brain injury rehabilitation. Ann Acad Med Singapore 2007; 36:31-42.

2. Khorasani-Zavareh D, Mohammadi R, Khankeh HR, Laflamme L, Bikmoradi A, Haglund BJ. The requirements and challenges in preventing of road traffic injury in Iran. A qualitative study. BMC Public Health 2009; 9:486.

3. Aghakhani N, Azami M, Jasemi M, Khoshsima M, Eghtedar S, Rahbar N. Epidemiology of traumatic brain injury in urmia, iran. Iran Red Crescent Med J 2013; 15:173.

4. Johnson VE, Stewart JE, Begbie FD, Trojanowski JQ, Smith DH, Stewart W. Inflammation and white matter degeneration persist for years after a single traumatic brain injury. Brain 2013; 136:28-42.

5. Ding K, Wang H, Xu J, Li T, Zhang L, Ding Y, et al. Melatonin stimulates antioxidant enzymes and reduces oxidative stress in experimental traumatic brain injury: the Nrf2-ARE signaling pathway as a potential mechanism. Free Radic Biol Med 2014; 73:1-11.

6. Bayir H, Kochanek PM, Clark RS. Traumatic brain injury in infants and children: mechanisms of secondary damage and treatment in the intensive care unit. Crit Care Clin 2003; 19:529-549.

7. Kunz A, Dirnagl U, Merqenthaler P. Acute pathophysiological processes after ischaemic and traumatic brain injury. Best Pract Res Clin Anaesthesiol 2010; 24:495-509.

8. Hall ED, Vaishnav RA, Mustafa AG. Antioxidant therapies for traumatic brain injury. Neurothera-peutics 2010; 7:51-61.

9. Reiter RJ. Melatonin: the chemical expression of darkness. Mol Cell Endocrinol 1991; 79:C153-158.

10. Claustrat B, Brun J, Chazot G. The basic physiology and pathophysiology of melatonin. Sleep Med Rev 2005; 9:11-24.

11. Seifman MA, Gomes K, Nguyen PN, Bailey M, Rosenfeld JV, Cooper DJ, et al. Measurement of serum melatonin in intensive care unit patients: changes in traumatic brain injury, trauma, and medical conditions. Front Neurol 2014; 5:273.

12. Ahmadiasl N, Shokofeh B, Alireza A. Combination Antioxidant Effect of erythropoietin and melatonin on renal ischemia-reperfusion injury in rats. Iran J Basic Med Sci 2013; 16:1209-1211.

13. Pineau I, Sun L, Bastien D, Lacroix S. Astrocytes initiate inflammation in the injured mouse spinal cord by promoting the entry of neutrophils and inflammatory monocytes in an IL-1 receptor/MyD88-dependent fashion. Brain Behav Immun 2010; 24:540-553.

14. Barreto GE, Gonzalez J, Torres Y, Morales L. Astrocytic-neuronal crosstalk: Implications for neuroprotection from brain injury. Neurosci Res 2011; 71:107-113.

15. Cameron B, Landreth GE. Inflammation, microglia, and alzheimer's disease. Neurobiol Dis 2010; 37:503-509.

16. Candace LF, Bruce GL. Astroglia: Important mediators of traumatic brain injury. Prog Brain Res 2007; 161.

17. Skaper SD, Floreani M, Ceccon M, Facci L, Giusti P. Excitotoxicity, oxidative stress, and the neuroprotective potential of melatonin. Ann N Y Acad Sci 1999; 890:107-118.

18. Dehghan F, Hadad MK, Asadikram G, Najafipour H, Shahrokhi N. Effect of melatonin on intracranial pressure and brain edema following traumatic brain injury: role of oxidative stresses. Arch Med Res 2013; 44:251-258.

19. A Marmarou, MAAE Foda, W Brink, J Campbell.  A new model of diffuse brain injury in rats: Part I: Pathophysiology and biomechanics. J Neurosurg 1994; 80:291-300.

20. Keshavarzi Z, Khaksari M, Shahrokhi N. The effects of cyclooxygenase inhibitors on the gastric emptying and small intestine transit in the male rats following traumatic brain injury. Iran J Basic Med Sci 2014; 17:406-410.

21. Koizumi S, Shigemoto-Mogami Y, Nasu-Tada K, Shinozaki Y, Ohsawa K, Tsuda M, et al. UDP acting at P2Y6 receptors is a mediator of microglial phagocytosis. Nature 2007; 446:1091-1095.

22. Ding K, Wang H, Xu J, Li T, Zhang L, Ding Y, et al. Melatonin stimulates antioxidant enzymes and reduces oxidative stress in experimental traumatic brain injury: the Nrf2-ARE signaling pathway as a potential mechanism. Free Radic Biol Med 2014; 73:1-11.

23. Lee MY, Kuan YH, Chen HY, Chen TY, Chen ST, Huang CC, et al. Intravenous administration of melatonin reduces the intracerebral cellular inflammatory response following transient focal cerebral ischemia in rats. J Pineal Res 2007; 42:297-309.

24. Ahmad molai Gh, Dabiri, Sh. Asadi karam GM, Shahrokhi N. Comparision of the effect of progestrone, allopregnanolone and gender on suppressing edema formation after traumatic brain injury in rat. Kerman Univ Med Sci 2008; 15:47-59.

25. Baydas G, Reiter RJ, Yasar A, Tuzcu M, Akdemir I, Nedzvetskii VS. Melatonin reduces glial reactivity in the hippocampus, cortex, and cerebellum of streptozotocin-induced diabetic rats. Free Radic Biol Med 2003; 35:797-804.

26. Li Y, Chopp M, Jiang N, Zhang ZG, Zaloga C. Induction of DNA fragmentation after 10 to 120 minutes of focal cerebral ischemia in rats. Stroke 1995; 26:1252-1258.

27. Kabadi SV, Maher TJ. Posttreatment with uridine and melatonin following traumatic brain injury reduces edema in various brain regions in rats. Ann N Y Acad Sci 2010; 1199:105–113.

28. Chern CM, Liao JF, Wang YH, Shen YC. Melatonin ameliorates neural function by promoting endogenous neurogenesis through the MT2 melatonin receptor in ischemic-stroke mice. Free Radic Biol Med 2012; 52:1634–1647.

29. Dehghan F, Khaksari Hadad M, Asadikram G, Najafipour H, Shahrokhi N. Effect of melatonin on intracranial pressure and brain edema following traumatic brain injury: role of oxidative stresses. Arch Med Res 2013; 44:251-258.

30. Ozdemir D, Uysal N, Gonenc S, Acikgoz O, Sonmez
A, Topcu A, et al. Effect of melatonin on brain oxidative damage induced by traumatic brain injury in immature rats. Physiol Res 2005; 54:631-637.

31. Welin AK, Svedin P, Lapatto R, Sultan B, Hagberg H, Gressens P, et al. Melatonin reduces inflammation and cell death in white matter in the mid-gestation fetal sheep following umbilical cord occlusion. Pediatr Res 2007; 61:153-158.

32. Keskin I, Kaplan S, Kalkan S, Sutcu M, Ulkay MB, Esener OB. Evaluation of neuroprotection by melatonin against adverse effects of prenatal exposure to a nonsteroidal anti-inflammatory drug during peripheral nerve development. Int J Dev Neurosci 2015; 41:1-7.

33. Tsai MC, Chen WJ, Tsai MS, Ching CH, Chuang JI. Melatonin attenuates brain contusion-induced oxidative insult, inactivation of signal transducers and activators of transcription 1, and upregulation of suppressor of cytokine signaling-3 in rats. J Pineal Res 2011; 51:233-245.

34. Marmarou CR, Liang X, Abidi NH, Parveen S, Taya K, Henderson SC, et al. Selective vasopressin-1a receptor antagonist prevents brain edema, reduces astrocytic cell swelling and GFAP, V1aR and AQP4 expression after focal traumatic brain injury. Brain Res 2014; 1581:89-102.

35. Pekny M, Nilsson M. Astrocyte activation and reactive gliosis. Glia 2005; 50:427-434.

36. Babaei-Balderlou F, Zare S, Heidari R, Farrokhi F. Effects of melatonin and vitamin E on peripheral neuropathic pain in streptozotocin-induced diabetic rats. Iran J Basic Med Sci 2010; 13:1-8.

37. Ananth C, Gopalakrishnakone P, Kaur C. Protective role of melatonin in domoic acid-induced neuronal damage in the hippocampus of adult rats. Hippocampus 2003; 13:375-387.

38. Abbott NJ, Patabendige AA, Dolman DE, Yusof SR, Begley DJ. Structure and function of the blood-brain barrier. Neurobiol Dis 2010; 37:13-25.

39. Ziebell JM, Morganti-Kossmann MC. Involvement of pro- and anti-inflammatory cytokines and chemokines in the pathophysiology of traumatic brain injury. Neurotherapeutics 2010; 7:22-30.

40. Domowicz MS, Henry JG, Wadlington N, Navarro A, Kraig RP. Astrocyte precursor response to embryonic brain injury. Brain Res 2011; 1389:35-49.

41. Ding K, Wang H, Xu J, Lu X, Zhang L, Zhu L. Melatonin reduced microglial activation and alleviated neuroinflammation induced neuron degeneration in experimental traumatic brain injury: Possible involvement of mTOR pathway. Neurochem Int 2014; 76:23-31.