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Diabetes enhances oxidative stress-induced TRPM2 channel activity and its control by N-acetylcysteine in rat dorsal root ganglion and brain

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Abstract

N-acetylcysteine (NAC) is a sulfhydryl donor antioxidant that contributes to the regeneration of glutathione (GSH) and also scavengers via a direct reaction with free oxygen radicals. Recently, we observed a modulatory role of NAC on GSH-depleted dorsal root ganglion (DRG) cells in rats. NAC may have a protective role on oxidative stress and calcium influx through regulation of the TRPM2 channel in diabetic neurons. Therefore, we investigated the effects of NAC on DRG TRPM2 channel currents and brain oxidative stress in streptozotocin (STZ)-induced diabetic rats. Thirty-six rats divided into four groups: control, STZ, NAC and STZ + NAC. Diabetes was induced in the STZ and STZ + NAC groups by intraperitoneal STZ (65 mg/kg) administration. After the induction of diabetes, rats in the NAC and STZ + NAC groups received NAC (150 mg/kg) via gastric gavage. After 2 weeks, DRG neurons and the brain cortex were freshly isolated from rats. In whole-cell patch clamp experiments, TRPM2 currents in the DRG following diabetes induction with STZ were gated by H2O2. TRPM2 channel current densities in the DRG and lipid peroxidation levels in the DRG and brain were higher in the STZ groups than in controls; however, brain GSH, GSH peroxidase (GSH-Px), vitamin C and vitamin E concentrations and DRG GSH-Px activity were decreased by diabetes. STZ + H2O2-induced TRPM2 gating was totally inhibited by NAC and partially inhibited by N-(p-amylcinnamoyl) anthranilic acid (ACA) and 2-aminoethyl diphenylborinate (2-APB). GSH-Px activity and lipid peroxidation levels were also attenuated by NAC treatment. In conclusion, we observed a modulatory role of NAC on oxidative stress and Ca2+ entry through the TRPM2 channel in the diabetic DRG and brain. Since excessive oxidative stress and overload Ca2+ entry are common features of neuropathic pain, our findings are relevant to the etiology and treatment of pain neuropathology in DRG neurons.

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Abbreviations

2-APB:

2-aminoethyl diphenylborinate

ACA:

N-(p-amylcinnamoyl) anthranilic acid

ADPR:

Adenosine diphosphatase ribose

DRG:

Dorsal root ganglion

EGTA:

Ethylene glycol-bis[2-aminoethyl-ether]-N,N,N,N-tetraacetic acid

GSH:

Reduced glutathione

GSH-Px:

Glutathione peroxidase

IP:

Intraperitoneal

LP:

Lipid peroxidation

NAC:

N-acetylcysteine

NMDG+ :

N-methyl-D-glucamine

ROS:

Reactive oxygen species

SDU:

Suleyman Demirel Universy

TRP:

Transient receptor potential

TRPM2:

Transient receptor potential melastatin 2

W.C.:

Whole cell

References

  • Amador-Alvarado L, Montiel T, Massieu L (2014) Differential production of reactive oxygen species in distinct brain regions of hypoglycemic mice. Metab Brain Dis 29:711–719

    Article  CAS  PubMed  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  PubMed Central  Google Scholar 

  • Butterfield DA, Di Domenico F, Barone E (2014) Elevated risk of type 2 diabetes for development of Alzheimer disease: a key role for oxidative stress in brain. Biochim Biophys Acta 1842:1693–1706

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Desai ID (1984) Vitamin E analysis methods for animal tissues. Methods Enzymol 105:138–147

    Article  CAS  PubMed  Google Scholar 

  • Dringen R (2000) Metabolism and functions of glutathione in brain. Prog Neurobiol 62:649–671

    Article  CAS  PubMed  Google Scholar 

  • Duzhyy DE, Viatchenko-Karpinski VY, Khomula EV, Voitenko NV, Belan PV (2015) Upregulation of T-type Ca2+ channels in long-term diabetes determines increased excitability of a specific type of capsaicin-insensitive DRG neurons. Mol Pain 11:29

    Article  PubMed  PubMed Central  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 29:787–799

    Article  CAS  PubMed  Google Scholar 

  • Jagota SK, Dani HM (1982) A new colorimetric technique for the estimation of vitamin C using folin phenol reagent. Anal Biochem 127:178–182

    Article  CAS  PubMed  Google Scholar 

  • Kahya MC, Nazıroğlu M, Çiğ B. (2015) Melatonin and selenium reduce plasma cytokine and brain oxidative stress levels in diabetic rats. Brain Inj 5. doi:10.3109/02699052.2015

  • Kamboj SS, Sandhir R (2011) Protective effect of N-acetylcysteine supplementation on mitochondrial oxidative stress and mitochondrial enzymes in cerebral cortex of streptozotocin-treated diabetic rats. Mitochondrion 11:214–222

    Article  CAS  PubMed  Google Scholar 

  • Köse SA, Nazıroğlu M (2015) N-acetyl cysteine reduces oxidative toxicity, apoptosis, and calcium entry through TRPV1 channels in the neutrophils of patients with polycystic ovary syndrome. Free Radic Res 49:338–346

    Article  PubMed  Google Scholar 

  • Kraft R, Grimm C, Frenzel H, Harteneck C (2006) Inhibition of TRPM2 cation channels by N-(p-amylcinnamoyl)anthranilic acid. Br J Pharmacol 148:264–273

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lawrence RA, Burk RF (1976) Glutathione peroxidase activity in selenium-deficient rat liver. Biochem Biophys Res Commun 71:952–958

    Article  CAS  PubMed  Google Scholar 

  • Lee M, Cho T, Jantaratnotai N, Wang YT, McGeer E, McGeer PL (2010) Depletion of GSH in glial cells induces neurotoxicity: relevance to aging and degenerative neurological diseases. Faseb j 24:2533–2545

    Article  CAS  PubMed  Google Scholar 

  • Lu S, Xiang L, Clemmer JS, Mittwede PN, Hester RL (2014) Oxidative stress increases pulmonary vascular permeability in diabetic rats through activation of transient receptor potential melastatin 2 channels. Microcirculation 21:754–760

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • McHugh D, Flemming R, Xu SZ, Perraud AL, Beech DJ (2003) Critical intracellular Ca2+ dependence of transient receptor potential melastatin 2 (TRPM2) cation channel activation. J Biol Chem. 278(13):11002–11006

    Article  CAS  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, Butterworth PJ, Sonmez TT (2011a) Dietary vitamin C and E modulates antioxidant levels in blood, brain, liver, muscle, and testes in diabetic aged rats. Int J Vitam Nutr Res 81:347–357

    Article  PubMed  Google Scholar 

  • Nazıroğlu M, Çelik Ö, Özgül C, Çiğ B, Doğan S, Bal R, Gümral N, Rodríguez AB, Pariente JA (2012b) Melatonin modulates wireless (2.45 GHz)-induced oxidative injury through TRPM2 and voltage gated Ca(2+) channels in brain and dorsal root ganglion in rat. Physiol Behav 105:683–692

    Article  PubMed  Google Scholar 

  • Nazıroğlu M, Ciğ B, Ozgül C (2013b) 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, Çiğ B, Özgül C (2014b) 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 

  • Nazıroğlu M, Dikici DM, Dursun S (2012a) 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, Özgül C, Çiğ B, Doğan S, Uğuz AC (2011b) Glutathione modulates Ca(2+) influx and oxidative toxicity through TRPM2 channel in rat dorsal root ganglion neurons. J Membr Biol 242:109–118

    Article  PubMed  Google Scholar 

  • Nazıroğlu M, Senol N, Ghazizadeh V, Yürüker V (2014a) (2014b) neuroprotection induced by N-acetylcysteine and selenium against traumatic brain injury-induced apoptosis and calcium entry in hippocampus of rat. Cell Mol Neurobiol 34:895–903

    Article  PubMed  Google Scholar 

  • Nazıroğlu M, Uğuz AC, Ismailoğlu Ö, Çiğ B, Özgül C, Borcak M (2013a) 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 (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 (2009) Role of selenium on calcium signaling and oxidative stress-induced molecular pathways in epilepsy. Neurochem Res 34:2181–2191

    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 (2015) Role of melatonin on calcium signaling and mitochondrial oxidative stress in epilepsy: focus on TRP channels. Tr J Biol 39:813–821

    Google Scholar 

  • Nesuashvili L, Hadley SH, Bahia PK, Taylor-Clark TE (2013) Sensory nerve terminal mitochondrial dysfunction activates airway sensory nerves via transient receptor potential (TRP) channels. Mol Pharmacol 83:1007–1019

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Övey IS, Nazıroğlu M (2015) Homocysteine and cytosolic GSH depletion induce apoptosis and oxidative toxicity through cytosolic calcium overload in the hippocampus of aged mice: involvement of TRPM2 and TRPV1 channels. Neuroscience 284:225–233

    Article  PubMed  Google Scholar 

  • Özdemir ÜS, Nazıroğlu M, Şenol N, Ghazizadeh V. (2015) Hypericum perforatum attenuates spinal cord injury-induced oxidative stress and apoptosis in the dorsal root ganglion of rats: Involvement of TRPM2 and TRPV1 Channels. Mol Neurobiol 23. doi:10.1007/s12035-015-9292-1

  • Özgül C, Nazıroğlu M (2012) TRPM2 channel protective properties of N-acetylcysteine on cytosolic glutathione depletion dependent oxidative stress and Ca2+ influx in rat dorsal root ganglion. Physiol Behav 106:122–128

    Article  PubMed  Google Scholar 

  • Özkaya D, Nazıroğlu M, Armağan A, Demirel A, Köroglu BK, Çolakoğlu N, Kükner A, Sönmez TT (2011) Dietary vitamin C and E modulates oxidative stress induced-kidney and lens injury in diabetic aged male rats through modulating glucose homeostasis and antioxidant systems. Cell Biochem Funct 29:287–293

    Article  PubMed  Google Scholar 

  • Placer ZA, Cushman L, Johnson BC (1966) Estimation of products of lipid peroxidation (malonyl dialdehyde) in biological fluids. Anal Biochem 16:359–364

    Article  CAS  PubMed  Google Scholar 

  • Sedlak J, Lindsay RH (1968) Estimation of total, protein-bound, and nonprotein sulfhydryl groups in tissue with ellman's reagent. Anal Biochem 25:192–205

    Article  CAS  PubMed  Google Scholar 

  • Sen CK, Packer L (2000) Thiol homeostasis and supplements in physical exercise. Am J Clin Nutr 72(2 Suppl):653S–669S

    CAS  PubMed  Google Scholar 

  • Senol 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  CAS  PubMed  Google Scholar 

  • Staaf S, Franck MC, Marmigère F, Mattsson JP, Ernfors P (2010) Dynamic expression of the TRPM subgroup of ion channels in developing mouse sensory neurons. Gene Expr Patterns 10:65–74

    Article  CAS  PubMed  Google Scholar 

  • Suzuki J, Katoh N (1990) A simple and cheap method for measuring vitamin a in cattle using only a spectrophotometer. Jpn J Vet Sci 52:1282–1284

    Google Scholar 

  • Uchida K, Tominaga M (2014) The role of TRPM2 in pancreatic β-cells and the development of diabetes. Cell Calcium 56(5):332–339

    Article  CAS  PubMed  Google Scholar 

  • Wehage E, Eisfeld J, Heiner I, Jüngling E, Zitt C, Lückhoff A (2002) Activation of the cation channel long transient receptor potential channel 2 (LTRPC2) by hydrogen peroxide. A splice variant reveals a mode of activation independent of ADP-ribose. J Biol Chem. 277(26):23150–23156

    Article  CAS  PubMed  Google Scholar 

  • Zochodne DW (1999) Diabetic neuropathies: features and mechanisms. Brain Pathol 9:369–391

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

MN formulated the present hypothesis and was responsible for writing the report. ES was responsible for the patch-clamp experiments, diabetes induction and treatments of animals. The authors wish thanks to Fatih Şahin and Muhammet Şahin (Department of Biophysics, Suleyman Demirel University) for helping the patch-clamp, lipid peroxidation and antioxidant analyses in this study. There are no conflicts of interest in the current study. The study was supported by Scientific Project Unit of SDU (Protocol No: BAP-3169-D2-12). All authors approved the final manuscript.

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Authors and Affiliations

  1. Department of Biophysics, Faculty of Medicine, Suleyman Demirel University, Isparta, Turkey

    Ercan Sözbir & Mustafa Nazıroğlu

  2. Neuroscience Research Center, Suleyman Demirel University, Isparta, Turkey

    Mustafa Nazıroğlu

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  1. Ercan Sözbir
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  2. Mustafa Nazıroğlu
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Correspondence to Mustafa Nazıroğlu.

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Sözbir, E., Nazıroğlu, M. Diabetes enhances oxidative stress-induced TRPM2 channel activity and its control by N-acetylcysteine in rat dorsal root ganglion and brain. Metab Brain Dis 31, 385–393 (2016). https://doi.org/10.1007/s11011-015-9769-7

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