Abdülhadi Cihangir UĞUZ, Abdülhadi Cihangir UĞUZ, Ahmi ÖZ, Ahmi ÖZ, Büşra YILMAZ, Seda ALTUNBAŞ, Ömer ÇELİK

Melatonin attenuates apoptosis and mitochondrial depolarization levels in hypoxic conditions of SH-SY5Y neuronal cells induced by cobalt chloride (CoCl2)

Melatonin (MEL) and its metabolites serve as endogenous reactive oxygen species (ROS) scavengers and have a wide spectrum of antioxidant activity. Cobalt chloride is one of the commonly used hypoxia-mimetic agents, due to blocking the degradation of and triggering the accumulation of hypoxia-inducible factor-1 alpha (HIF-1α ) protein, which is very well known as a critical regulator of the cellular response against hypoxia. In the current study we aimed to determine the possible protective effects of melatonin on a cobalt chloride-induced hypoxia model of SH-SY5Y neuronal cells. Group I was the control group and SH-SY5Y cells were incubated in normal culture media without any chemical administration. In Group II, SH-SY5Y cells were incubated with 1 μM MEL for 24 h. In Group III cells were incubated with 200 μM cobalt chloride for 24 h. The last group, Group 4, was a combination group of cobalt chloride and MEL. Cells were preincubated with 1 μM for 12 h and then 200 μM cobalt chloride for 24 h. We performed the cell viability test (MTT) and checked the caspase-3 and -9 activities and the lipid peroxidation (LP), reduced glutathione (GSH), glutathione peroxidase (GSH-Px), mitochondrial depolarization, and intracellular ROS levels. We observed that cobalt chloride increased intracellular ROS, caspase-3 and -9, lipid peroxidation, and mitochondrial membrane depolarization values, while decreasing GSH levels and GSH-Px activity. However, GSH and GSH-Px values were increased by melatonin treatment although lipid peroxidation level, intracellular ROS production, and caspase-3 and -9 activities were decreased by the treatment. In conclusion, we observed that melatonin incubation protects neuronal cells against hypoxia-induced oxidative stress.

Anahtar Kelimeler:

SH-SY5Y neuroblastoma cells, cobalt chloride, oxidative stress, melatonin, mitochondrial membrane depolarization

Melatonin attenuates apoptosis and mitochondrial depolarization levels in hypoxic conditions of SH-SY5Y neuronal cells induced by cobalt chloride (CoCl2)

Keywords:

SH-SY5Y neuroblastoma cells, cobalt chloride, oxidative stress, melatonin, mitochondrial membrane depolarization,

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Amoroso S, D’Alessio A, Sirabella R, Di Renzo G, Annunziato L (2002). Ca(2+)-independent caspase-3 but not Ca(2+)- dependent caspase-2 activation induced by oxidative stress leads to SH-SY5Y human neuroblastoma cell apoptosis. J Neurosci Res 68: 454–462.
Amoroso S, Gioielli A, Cataldi M, Di Renzo G, Annunziato L (1999). In the neuronal cell line SH-SY5Y, oxidative stress-induced free radical overproduction causes cell death without any participation of intracellular Ca(2+) increase. Biochim Biophys Acta 1452: 151–160.
Banno Y, Wang S, Ito Y, Izumi T, Nakashima S, Shimizu T, Nozawa Y (2001). Involvement of ERK and p38 MAP kinase in oxidative stress-induced phospholipase D activation in PC12 cells. Neuroreport 12: 2271–2275.
Barrett P, Bolborea M (2012). Molecular pathways involved in seasonal body weight and reproductive responses governed by melatonin. J Pineal Res 52: 376–388.
Bejarano I, Redondo PC, Espino J, Rosado JA, Paredes SD, Barriga C, Reiter RJ, Pariente JA, Rodriguez AB (2009). Melatonin induces mitochondrial-mediated apoptosis in human myeloid HL-60 cells. J Pineal Res 46: 392–400.
Carrillo-Vico A, Garcia-Maurino S, Calvo JR, Guerrero JM (2003). Melatonin counteracts the inhibitory effect of PGE2 on IL-2 production in human lymphocytes via its mt1 membrane receptor. FASEB J 17: 755–757.
Chang CY, Lui TN, Lin JW, Lin YL, Hsing CH, Wang JJ, Chen RM (2014). Roles of microRNA-1 in hypoxia-induced apoptotic insults to neuronal cells. Arch Toxicol (in press).
Chen L, Liu L, Yin J, Luo Y, Huang S (2009). Hydrogen peroxideinduced neuronal apoptosis is associated with inhibition of protein phosphatase 2A and 5, leading to activation of MAPK pathway. Int J Biochem Cell Biol 41: 1284–1295.
Chen TI, Chiu HW, Pan YC, Hsu ST, Lin JH, Yang KT (2014). Intermittent hypoxia-induced protein phosphatase 2A activation reduces PC12 cell proliferation and differentiation. J Biomed Sci 21: 46.
D’Amelio M, Cavallucci V, Cecconi F (2010). Neuronal caspase-3 signaling: not only cell death. Cell Death Differ 17: 1104–1114.
Das A, McDowell M, Pava MJ, Smith JA, Reiter RJ, Woodward JJ, Varma AK, Ray SK, Banik NL (2010). The inhibition of apoptosis by melatonin in VSC4.1 motoneurons exposed to oxidative stress, glutamate excitotoxicity, or TNF-alpha toxicity involves membrane melatonin receptors. J Pineal Res 48: 157– 169.
Dunn KC, Aotaki-Keen AE, Putkey FR, Hjelmeland LM (1996). ARPE-19, a human retinal pigment epithelial cell line with differentiated properties. Exp Eye Res 62: 155–169.
Espino J, Bejarano I, Paredes SD, Barriga C, Reiter RJ, Pariente JA, Rodriguez AB (2011a). Melatonin is able to delay endoplasmic reticulum stress-induced apoptosis in leukocytes from elderly humans. Age (Dordr) 33: 497–507.
Espino J, Bejarano I, Paredes SD, Barriga C, Rodriguez 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.
Gaitanaki C, Kalpachidou T, Aggeli IK, Papazafiri P, Beis I (2007). CoCl2 induces protective events via the p38-MAPK signaling pathway and ANP in the perfused amphibian heart. J Exp Biol 210: 2267–2277.
Galano A, Tan DX, Reiter RJ (2011). Melatonin as a natural ally against oxidative stress: a physicochemical examination. J Pineal Res 51: 1–16.
Galano A, Tan DX, Reiter RJ (2013). On the free radical scavenging activities of melatonin’s metabolites, AFMK and AMK. J Pineal Res 54: 245–257.
Hill SM, Frasch T, Xiang S, Yuan L, Duplessis T, Mao L (2009). Molecular mechanisms of melatonin anticancer effects. Integr Cancer Ther 8: 337–346.
Lan A, Liao X, Mo L, Yang C, Yang Z, Wang X, Hu F, Chen P, Feng J, Zheng D et al. (2011). Hydrogen sulfide protects against chemical hypoxia-induced injury by inhibiting ROS-activated ERK1/2 and p38MAPK signaling pathways in PC12 cells. PLoS One 6: e25921.
Lawrence RA, Burk RF (1976). Glutathione peroxidase activity in selenium-deficient rat liver. Biochem Biophys Res Commun 71: 952–958.
Liu AL, Wang XW, Liu AH, Su XW, Jiang WJ, Qiu PX, Yan GM (2009). JNK and p38 were involved in hypoxia and reoxygenationinduced apoptosis of cultured rat cerebellar granule neurons. Exp Toxicol Pathol 61: 137–143.
Lowry OH, Rosenbrough NJ, Farr AL, Randall RJ (1951). Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265–275.
Martin M, Macias M, Escames G, Reiter RJ, Agapito MT, Ortiz GG, Acuna-Castroviejo D (2000). Melatonin-induced increased activity of the respiratory chain complexes I and IV can prevent mitochondrial damage induced by ruthenium red in vivo. J Pineal Res 28: 242–248.
Martin V, Sainz RM, Antolin I, Mayo JC, Herrera F, Rodriguez C (2002). Several antioxidant pathways are involved in astrocyte protection by melatonin. J Pineal Res 33: 204–212.
Naziroglu M, Cig B, Ozgul 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.
Olivieri G, Hess C, Savaskan E, Ly C, Meier F, Baysang G, Brockhaus M, Müller-Spahn F (2001). Melatonin protects SHSY5Y neuroblastoma cells from cobalt-induced oxidative stress, neurotoxicity, and increased beta-amyloid secretion. J Pineal Res. 31: 320–325.
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 (in press).
Paredes Royano SD, Reiter RJ (2010). Melatonin: helping cells cope with oxidative disaster. Cell Membr Free Radic Res 2: 99–111.
Placer ZA, Cushman LL, Johnson BC (1966). Estimation of product of lipid peroxidation (malonyl dialdehyde) in biochemical systems. Anal Biochem 16: 359–364.
Sakaida I, Thomas AP, Farber JL (1991). Increases in cytosolic calcium ion concentration can be dissociated from the killing of cultured hepatocytes by tert-butyl hydroperoxide. J Biol Chem 266: 717–722.
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.
Tan DX, Hardeland R, Manchester LC, Paredes SD, Korkmaz A, Sainz RM, Mayo JC, Fuentes-Broto L, Reiter RJ (2010). The changing biological roles of melatonin during evolution: from an antioxidant to signals of darkness, sexual selection and fitness. Biol Rev Camb Philos Soc 85: 607–623.
Tan DX, Manchester LC, Reiter RJ, Qi WB, Zhang M, Weintraub ST, Cabrera J, Sainz RM, Mayo JC (1999). Identification of highly elevated levels of melatonin in bone marrow: its origin and significance. Biochim Biophys Acta 1472: 206–214.
Uguz AC, Cig B, Espino J, Bejarano I, Naziroglu M, Rodriguez AB, Pariente JA (2012). Melatonin potentiates chemotherapyinduced cytotoxicity and apoptosis in rat pancreatic tumor cells. J Pineal Res 53: 91–98.
Uguz AC, Naziroglu 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.
Uguz AC, Naziroglu M, Espino J, Bejarano I, Gonzalez D, Rodriguez AB, Pariente JA (2009). Selenium modulates oxidative stressinduced cell apoptosis in human myeloid HL-60 cells through regulation of calcium release and caspase-3 and -9 activities. J Membr Biol 232: 15–23.
Yong-Kee CJ, Sidorova E, Hanif A, Perera G, Nash JE (2012). Mitochondrial dysfunction precedes other subcellular abnormalities in an in vitro model linked with cell death in Parkinson’s disease. Neurotox Res 21: 185–194.
Yurekli VA, Gurler S, Naziroglu M, Uguz AC, Koyuncuoglu HR (2013). Zonisamide attenuates MPP+-induced oxidative toxicity through modulation of Ca2+ signaling and caspase-3 activity in neuronal PC12 cells. Cell Mol Neurobiol 33: 205– 212.