Skip to main content

Advertisement

Log in

Reduction in traumatic brain injury-induced oxidative stress, apoptosis, and calcium entry in rat hippocampus by melatonin: Possible involvement of TRPM2 channels

Metabolic Brain Disease Aims and scope Submit manuscript

Abstract

Melatonin, which is a very effective reactive oxygen species (ROS) scavenger, acts through a direct reaction with free radicals. Ca2+ entry induced by traumatic brain injury (TBI) has deleterious effects on human hippocampal function. TRPM2 is a Ca2+ permeable non-selective channel in hippocampal neurons, and its activation of during oxidative stress has been linked to cell death. Despite the importance of oxidative stress in TBI, its role in apoptosis and Ca2+ entry in TBI is poorly understood. Therefore, we tested the effects of melatonin on apoptosis, oxidative stress, and Ca2+ entry through the TRPM2 channel in the hippocampal neurons of TBI-induced rats. Thirty-two rats were divided into the following four groups: control, melatonin, TBI, and TBI + melatonin groups. Melatonin (5 mg/kg body weight) was intraperitoneally given to animals in the melatonin group and the TBI + melatonin group after 1 h of brain trauma. Hippocampal neurons were freshly isolated from the four groups, incubated with a nonspecific TRPM2 blocker (2-aminoethyl diphenylborinate, 2-APB), and then stimulated with cumene hydroperoxide. Apoptosis, caspase-3, caspase-9, intracellular ROS production, mitochondrial membrane depolarization and intracellular free Ca2+ ([Ca2+]i) values were high in the TBI group, and low in the TBI + melatonin group. The [Ca2+]i concentration was decreased in the four groups by 2-APB. In our TBI experimental model, TRPM2 channels were involved in Ca2+ entry-induced neuronal death, and the negative modulation of the activity of this channel by melatonin pretreatment may account for the neuroprotective activity of TRPM2 channels against oxidative stress, apoptosis, and Ca2+ entry.

Possible molecular pathways of traumatic brain injury (TBI) on Ca2+ signaling, oxidative stress and apoptosis values through TRPM2 channels in hippocampal neurons. It is likely that TRPM2-mediated Ca2+ entry in the hippocampus of TBI-induced rats involves accumulation of ROS and opening of mitochondrial permeability transition (MPT) that consequently leads to mitochondrial dysfunction. When the low oxidative stress and Ca2+ are just enough to lead to the mitochondrial membrane pores, physiological cell functions may be induced. At the extreme, oxidative stress and Ca2+ entry cause severe MPT or even the rupture of the mitochondrial membrane, substantial swelling of the mitochondria with rupture of the outer membrane and release of apoptosis-inducing factors such as caspase-3 and -9.

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

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Abbreviations

[Ca2+](i):

Intracellular Ca2+

2-APB:

2-aminoethyl diphenylborinate

DMSO:

Dimethyl sulfoxide

MPT:

Mitochondrial permeability transition

ROS:

Reactive oxygen species

TBI:

Traumatic brain injury

TRP:

Transient receptor potential

TRPM2:

Melastatin-like transient receptor potential 2

VGCC:

Voltage gated calcium channels

SDU:

Suleyman Demirel University

References

  • Akpinar A, Uğuz AC, Nazıroğlu M (2014) Agomelatine and duloxetine synergistically modulates apoptotic pathway by inhibiting oxidative stress triggered intracellular calcium entry in neuronal PC12 cells: role of TRPM2 and voltage-gated calcium channels. J Membr Biol 247:451–459

    Article  CAS  PubMed  Google Scholar 

  • Argun M, Tök L, Uğuz AC, Celik O, Tök OY, Nazıroğlu M (2014) Melatonin and amfenac modulate calcium entry, apoptosis, and oxidative stress in ARPE-19 cell culture exposed to blue light irradiation (405 nm). Eye (Lond) 28(6):752–760

    Article  CAS  Google Scholar 

  • Belrose JC, Xie YF, Gierszewski LJ, MacDonald JF, Jackson MF (2012) Loss of glutathione homeostasis associated with neuronal senescence facilitates TRPM2 channel activation in cultured hippocampal pyramidal neurons. Mol Brain 5:11

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Cabrera J, Reıter R, Tan DX, Qi W, Saniz RM, Mayo JC, Garcia JJ, Kim SJ, El-Sokkary G (2000) Melatonin reduces oxidative neurotoxicity due to quinolinic acid: in vitro and in vivo findings. Neuropharmacology 39:507–514

    Article  CAS  PubMed  Google Scholar 

  • Carrasco C, Marchena AM, Holguín-Arévalo MS, Martín-Partido G, Rodríguez AB, Paredes SD, Pariente JA (2013) Anti-inflammatory effects of melatonin in a rat model of caerulein-induced acute pancreatitis. Cell Biochem Funct 31:585–590

    CAS  PubMed  Google Scholar 

  • Celik O, Nazıroğlu M (2012) Melatonin modulates apoptosis and TRPM2 channels in transfected cells activated by oxidative stress. Physiol Behav 107:458–465

    Article  CAS  PubMed  Google Scholar 

  • Chang CF, Huang HJ, Lee HC, Hung KC, Wu RT, Lin AM (2012) Melatonin attenuates kainic acid-induced neurotoxicity in mouse hippocampus via inhibition of autophagy and α-synuclein aggregation. J Pineal Res 52:312–321

    Article  CAS  PubMed  Google Scholar 

  • Cook NL, Vink R, Helps SC, Manavis J, van den Heuvel C (2010) Transient receptor potential melastatin 2 expression is increased following experimental traumatic brain injury in rats. J Mol Neurosci 42:192–199

    Article  CAS  PubMed  Google Scholar 

  • Dilek M, Naziroğlu M, Baha Oral H, Suat Ovey I, Küçükayaz M, Mungan MT, Kara HY, Sütçü R (2010) Melatonin modulates hippocampus NMDA receptors, blood and brain oxidative stress levels in ovariectomized rats. J Membr Biol 233:135–142

    Article  CAS  PubMed  Google Scholar 

  • Ekmekcioglu C (2006) Melatonin receptors in humans: biological role and clinical relevance. Biomed Pharmaceut 60:97–108

    Article  CAS  Google Scholar 

  • Erşahin M, Özdemir Z, Özsavcı D, Akakın D, Yeğen B, Reiter RJ, Sener G (2012) Melatonin treatment protects against spinal cord injury induced functional and biochemical changes in rat urinary bladder. J Pineal Res 52:340–348

    Article  PubMed  Google Scholar 

  • Espino J, Bejarano I, Redondo PC, Rosado JA, Barriga C, Reiter RJ, Pariente JA, Rodríguez AB (2010) Melatonin reduces apoptosis induced by calcium signaling in human leukocytes: evidence for the involvement of mitochondria and Bax activation. J Membr Biol 233:105–118

    Article  CAS  PubMed  Google Scholar 

  • Espino J, Bejarano I, Paredes SD, Barriga C, Reiter RJ, Pariente JA, Rodríguez AB (2011a) Melatonin is able to delay endoplasmic reticulum stress-induced apoptosis in leukocytes from elderly humans. Age (Dordr) 33:497–507

    Article  CAS  Google Scholar 

  • Espino J, Bejarano I, Paredes SD, Barriga C, Rodríguez AB, Pariente JA (2011b) Protective effect of melatonin against human leukocyte apoptosis induced by intracellular calcium overload: relation with its antioxidant actions. J Pineal Res 51:195–206

    Article  CAS  PubMed  Google Scholar 

  • Espino J, Rodríguez AB, Pariente JA (2013) The inhibition of TNF-α-induced leucocyte apoptosis by melatonin involves membrane receptor MT1/MT2 interaction. J Pineal Res 54:442–452

    CAS  PubMed  Google Scholar 

  • Esposito E, Cuzzocrea S (2010) Antiinflammatory activity of melatonin in central nervous system. Curr Neuropharmacol 8:228–242

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Ghazizadeh V, Nazıroğlu M (2014) Electromagnetic radiation (Wi-Fi) and epilepsy induce calcium entry and apoptosis through activation of TRPV1 channel in hippocampus and dorsal root ganglion of rats. Metab Brain Dis

  • Grynkiewicz C, Poenie M, Tsien RY (1985) A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem 260:3440–3450

    CAS  PubMed  Google Scholar 

  • Gurkoff G, Shahlaie K, Lyeth B, Berman R (2013) Voltage-gated calcium channel antagonists and traumatic brain injury. Pharmaceuticals (Basel) 6:788–812

    Article  CAS  Google Scholar 

  • Iuvone PM, Boatright JH, Tosini G, Ye K (2014) N-acetylserotonin: circadian activation of the BDNF receptor and neuroprotection in the retina and brain. Adv Exp Med Biol 801:765–771

    Article  PubMed Central  PubMed  Google Scholar 

  • Maharaj DS, Maharaj H, Daya S, Glass BD (2006) Melatonin and 6-hydroxymelatonin protect against iron-induced neurotoxicity. J Neurochem 96:78–81

    Article  CAS  PubMed  Google Scholar 

  • Marmarou A, Foda MA, Brink WVB, Campbell J, Kita H, Demetriadou K (1994) A new model of diffuse brain injury in rats. J Neurosurg 80:291–300

    Article  CAS  PubMed  Google Scholar 

  • Nazıroğlu M (2007) New molecular mechanisms on the activation of TRPM2 channels by oxidative stress and ADP-ribose. Neurochem Res 32:1990–2001

    Article  PubMed  Google Scholar 

  • Nazıroğlu M (2011) TRPM2 cation channels, oxidative stress and neurological diseases: where are we now? Neurochem Res 36:355–366

    Article  PubMed  Google Scholar 

  • Nazıroğlu M, Lückhoff A (2008a) Effects of antioxidants on calcium influx through TRPM2 channels in transfected cells activated by hydrogen peroxide. J Neurol Sci 270:152–158

    Article  PubMed  Google Scholar 

  • Nazıroğlu M, Lückhoff A (2008b) A calcium influx pathway regulated separately by oxidative stress and ADP-Ribose in TRPM2 channels: single channel events. Neurochem Res 33:1256–1262

    Article  PubMed  Google Scholar 

  • Nazıroğlu M, Lückhoff A, Jüngling E (2007) Antagonist effect of flufenamic acid on TRPM2 cation channels activated by hydrogen peroxide. Cell Biochem Funct 25:383–387

    Article  PubMed  Google Scholar 

  • Nazıroğlu M, Çelik Ö, Özgül C, Doğan S, Bal R, Gümral N, Rodríguez AB, Pariente JA (2012a) Melatonin modulates wireless devices (2.45 GHz)-induced brain and dorsal root ganglion injury through TRPM2 and voltage gated calcium channels in rat. Physiol Behav 105:683–692

    Article  PubMed  Google Scholar 

  • Nazıroğlu M, Dikici DM, Dursun S (2012b) Role of oxidative stress and Ca(2+) signaling on molecular pathways of neuropathic pain in diabetes: focus on TRP channels. Neurochem Res 37:2065–2075

    Article  PubMed  Google Scholar 

  • Nazıroğlu M, Tokat S, Demirci S (2012c) Role of melatonin on electromagnetic radiation-induced oxidative stress and Ca2+ signaling molecular pathways in breast cancer. J Recept Signal Transduct Res 32:290–297

    Article  PubMed  Google Scholar 

  • Nazıroğlu M, Ciğ B, Ozgül C (2013a) Neuroprotection induced by N-acetylcysteine against cytosolic glutathione depletion-induced Ca2+ influx in dorsal root ganglion neurons of mice: role of TRPV1 channels. Neuroscience 242:151–160

    Article  PubMed  Google Scholar 

  • Nazıroğlu M, Uğuz AC, Ismailoğlu Ö, Çiğ B, Özgül C, Borcak M (2013b) Role of TRPM2 cation channels in dorsal root ganglion of rats after experimental spinal cord injury. Muscle Nerve 48:945–950

    Article  PubMed  Google Scholar 

  • Nazıroğlu M, Çiğ B, Özgül C (2014) Modulation of oxidative stress and Ca(2+) mobilization through TRPM2 channels in rat dorsal root ganglion neuron by Hypericum perforatum. Neuroscience 263:27–35

    Article  PubMed  Google Scholar 

  • Ozdemir D, Tugyan K, Uysal N, Sonmez U, Sonmez A, Acikgoz O, Ozdemir N, Duman M, Ozkan H (2005) Protective effect of melatonin against head trauma-induced hippocampal damage and spatial memory deficits in immature rats. Neurosci Lett 385:234–239

    Article  CAS  PubMed  Google Scholar 

  • Pandi-Perumal SR, BaHammam AS, Brown GM, Spence DW, Bharti VK, Kaur C, Hardeland R, Cardinali DP (2013) Melatonin antioxidative defense: therapeutical implications for aging and neurodegenerative processes. Neurotox Res 23:267–300

    Article  CAS  PubMed  Google Scholar 

  • Paredes SD, Reiter RJ (2010) Melatonin: helping cells cope with oxidative disaster. Cell Membr Free Radic Res 2:99–111

    Google Scholar 

  • Paredes SD, Terrón MP, Marchena AM, Barriga C, Pariente JA, Reiter RJ, Rodríguez AB (2007) Effect of exogenous melatonin on viability, ingestion capacity, and free-radical scavenging in heterophils from young and old ringdoves (Streptopelia risoria). Mol Cell Biochem 304:305–314

    Article  CAS  PubMed  Google Scholar 

  • Park K, Lee Y, Park S, Lee S, Hong Y, Kil Lee S, Hong Y (2010) Synergistic effect of melatonin on exercise-induced neuronal reconstruction and functional recovery in a spinal cord injury animal model. J Pineal Res 48:270–281

    Article  CAS  PubMed  Google Scholar 

  • Park S, Lee SK, Park K, Lee Y, Hong Y, Lee S, Jeon JC, Kim JH, Lee SR, Chang KT, Hong Y (2012) Beneficial effects of endogenous and exogenous melatonin on neural reconstruction and functional recovery in an animal model of spinal cord injury. J Pineal Res 52:107–119

    Article  CAS  PubMed  Google Scholar 

  • Perraud AL, Fleig A, Dunn CA, Bagley LA, Launay P, Schmitz C, Stokes AJ, Zhu Q, Bessman MJ, Penner R, Kinet JP, Scharenberg AM (2001) ADP-ribose gating of the calcium-permeable LTRPC2 channel revealed by Nudix motif homology. Nature 411:595–599

    Article  CAS  PubMed  Google Scholar 

  • Ray SK, Dixon CE, Banik NL (2002) Molecular mechanisms in the pathogenesis of traumatic brain injury. Histol Histopathol 17:1137–1152

    CAS  PubMed  Google Scholar 

  • Reiter RJ, Tan DX, Osuna C, Gitto E (2000) Actions of melatonin in the reduction of oxidative stress. A review. J Biomed Sci 7:444–458

    Article  CAS  PubMed  Google Scholar 

  • Samantaray S, Sribnick EA, Das A, Knaryan VH, Matzelle DD, Yallapragada AV, Reiter RJ, Ray SK, Banik NL (2008) Melatonin attenuates calpain upregulation, axonal damage and neuronal death in spinal cord injury in rats. J Pineal Res 44:348–357

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Schäfer MK, Pfeiffer A, Jaeckel M, Pouya A, Dolga AM, Methner A (2014) Regulators of mitochondrial Ca(2+) homeostasis in cerebral ischemia. Cell Tissue Res

  • Şenol N, Nazıroğlu M (2014) Melatonin reduces traumatic brain injury-induced oxidative stress in the cerebral cortex and blood of rats. Neural Regen Res 9:1112–1116

    Article  PubMed Central  PubMed  Google Scholar 

  • Şenol N, Nazıroğlu M, Yürüker V (2014) N-acetylcysteine and selenium modulate oxidative stress, antioxidant vitamin and cytokine values in traumatic brain injury-induced rats. Neurochem Res 39:685–692

    Article  PubMed  Google Scholar 

  • Springer JE (2002) Apoptotic cell death following traumatic injury to the central nervous system. J Biochem Mol Biol 35:94–105

    Article  CAS  PubMed  Google Scholar 

  • Uğuz AC, Nazıroğlu M (2012) Effects of selenium on calcium signaling and apoptosis in rat dorsal root ganglion neurons induced by oxidative stress. Neurochem Res 37:1631–1638

    Article  PubMed  Google Scholar 

  • Uguz AC, Cig B, Espino J, Bejarano I, Naziroglu M, Rodríguez AB, Pariente JA (2012) Melatonin potentiates chemotherapy-induced cytotoxicity and apoptosis in rat pancreatic tumor cells. J Pineal Res 53:91–98

    Article  CAS  PubMed  Google Scholar 

  • Vitte PA, Harthe C, Lestage P, Claustrat B, Bobillier P (1988) Plasma, cerebrospinal fluid, and brain distribution of 14C-melatonin in rat: a biochemical and autoradiographic study. J Pineal Res 5:437–453

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

The study was partially supported by Scientific Research Project Unit of Suleyman Demirel University (BAP-3265-TU-2-12). The authors wish thanks to Technician Fatih Şahin (Neuroscience Research Center, Suleyman Demirel University) for excellent performance on the analyses of current study. Abstract of the study will be submitted in 5th International Congress on Cell Membranes and Oxidative Stress: Focus on Calcium Signaling and TRP Channels, 9–12 September 2014, Isparta Turkey. (http://www.cmos.org.tr/2014/).

Authors’ roles

MN and NŞ formulated the present hypothesis and MN was also responsible for writing the report. VY was responsible for induction of TBI.

Disclosures

None of the authors have any conflicts to disclose. All authors approved the final manuscript.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Mustafa Nazıroğlu.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Yürüker, V., Nazıroğlu, M. & Şenol, N. Reduction in traumatic brain injury-induced oxidative stress, apoptosis, and calcium entry in rat hippocampus by melatonin: Possible involvement of TRPM2 channels. Metab Brain Dis 30, 223–231 (2015). https://doi.org/10.1007/s11011-014-9623-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11011-014-9623-3

Keywords

Navigation