Regulatory Effects of Thymoquinone on Dopamine Level in Neuronal Cells Exposed to Amphetamine: An In Vitro Study
Journal of Cellular & Molecular Anesthesia,
Vol. 5 No. 4 (2020),
Introduction: Amphetamine (AT) is used to treat some medical conditions and also known to be abused recreationally. It is a potent central nervous system stimulant that is capable of producing damaging effects to the central dopaminergic pathway. Most of AT users are treated clinically for symptomatic treatment which is associated with neurological side effects. To date, there is growing interest in naturally occurring compounds which have lesser side effects to treat health problems. One of the potential compounds is thymoquinone (TQ), an active compound of Nigella sativa which is known for its cellular protective effects. Objective: The objectives of this study were to determine the IC50 values of AT and TQ on differentiated SH-SY5Y neuronal cells and to evaluate the changes of dopamine (DA) level in the cells exposed to AT after co-administering with TQ. Methodology: Differentiated SH-SY5Y cells were grown in cell culture flask containing DMEM/F12 medium supplemented with 10% (v/v) fetal bovine serum and 1% (v/v) penicillin/streptomycin. The IC50 value of TQ and AT in differentiated SH-SY5Y cells was determined by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The DA level was determined by using the Enzyme-Linked Immunosorbent Assay (ELISA) kit. Result and Discussion: The IC50 values of AT and TQ were 1596 µM and 926 µM respectively. Co-administration of 40 µM of AT and 30 µM of TQ demonstrated a significant increase in DA level at 48 hours of exposure when compared to the administration of AT group (P≤0.05). Conclusion: These findings suggested that TQ has a role in maintaining the DA activity after a long-term AT exposure.
- differentiated SH-SY5Y cells
- lactate dehydrogenase activity
How to Cite
Bramness JG, Gundersen OH, Guterstam J, et al. Amphetamine-induced psychosis-a separate diagnostic entity or primary psychosis triggered in the vulnerable? BMC Psychiatry. 2012;12(1):221-7.
Greene SL, Kerr F, Braitberg G. Amphetamines and related drugs of abuse. Emerg Med Australas. 2008;20(5):391-402.
Tzschentke TM. Pharmacology and behavioral pharmacology of the mesocortical dopamine system. Prog Neurobiol. 2001;63(3):241-320.
Uddin MS, Sufian MA, Kabir MT, Hossain MF, Nasrullah M, Islam L. Amphetamines: Potent recreational drug of abuse. J Addic Res The. 2017;8(4):1-12.
Schrantee A, Vaclavu L, Heijtel DF, et al. Dopaminergic system dysfunction in recreational dexamphetamine users. Neuropsychopharmacology. 2015; 40(5):1172-80.
Ciccarone D. Stimulant abuse: pharmacology, cocaine, methamphetamine, treatment, attempts at pharmacotherapy. Primary Care: Clinics in Office Practice. 2011;38(1):41-58.
Berman SM, Kuczenski R, McCracken JT, London ED. Potential adverse effects of amphetamine treatment on brain and behavior: a review. Mo Psychiatry. 2009;14(2):123-42.
Richards JR, Albertson TE, Derlet RW, Lange RA, Olson KR, Horowitz BZ. Treatment of toxicity from amphetamines, related derivatives, and analogues: a systematic clinical review. Drug and Alcohol Dependence. 2015;150:1-13.
Ayano G. First generation antipsychotics: pharmacokinetics, pharmacodynamics, therapeutic effects and side effects: A review. Research & Reviews: J Chemistry. 2016;5(3):53-63.
Sun HQ, Chen HM, Yang FD, Lu L, Kosten TR. Epidemiological trends and the advances of treatments of amphetamine‐type stimulants (ATS) in China. The American Journal on Addictions. 2014;23(3):313-7.
Zhang J, Wider B, Shang H. Li X, Ernst E. Quality of herbal medicines: challenges and solutions. Complementary Therapies in Medicine. 2012;20(1-2):100-6.
Hobbenaghi R, Javanbakht J, Sadeghzadeh S. et al. Neuroprotective effects of Nigella sativa extract on cell death in hippocampal neurons following experimental global cerebral ischemia-reperfusion injury in rats. J Neurol Sci. 2014;337(1-2):74-9.
Darakhshan S, Bidmeshki PA, Hosseinzadeh CA, Sisakhtnezhad S. Thymoquinone and its therapeutic potentials. Pharmacol Res. 2015;95:138-58.
Ghayur MN, Gilani AH, Janssen LJ. Intestinal, airway, and cardiovascular relaxant activities of thymoquinone. Evid-Based Compl Alt Med. 2012;2012:1-13.
Radad SK, Al-Shraim MM, Moustafa FM, Rausch W. Neuroprotective role of thymoquinone against 1-methyl-4-phenylpyridinium-induced dopaminergic cell death in primary mesencephalic cell culture. Neurosciences, 2015:20 (1):10-6.
Ahmad A, Husain A, Mujeeb M. Khan SA, Najmi AK, Siddique NA, et al. A review on therapeutic potential of Nigella sativa: A miracle herb. Asian Pac J Trop Biomed. 2013;3(5):337-52.
Beheshti F, Khazaei M, Hosseini M. Neuropharmacological effects of Nigella sativa. Avicenna J Phytomedicine. 2016;6(1):104-16.
Radad K, Moldzio R, Taha M, Rausch WD. Thymoquinone protects dopaminergic neurons against MPP+ and rotenone. Phytotherapy Research: An International Journal Devoted to Pharmacological and Toxicological Evaluation of Natural Product Derivatives. 2009;23(5):696-700.
Kassab RB, El-Hennamy RE. The role of thymoquinone as a potent antioxidant in ameliorating the neurotoxic effect of sodium arsenate in female rat. Egyptian Journal of Basic and Applied Sciences. 2017;4(3):160-7.
Presgraves SP, Ahmed T, Borwege S, Joyce JN. Terminally differentiated SH-SY5Y cells provide a model system for studying neuroprotective effects of dopamine agonists. Neurotox Res. 2003;5(8):579-98.
Cheung YT, Lau WKW, Yu MS. et al. Effects of all-trans-retinoic acid on human SH-SY5Y neuroblastoma as in vitro model in neurotoxicity research. Neurotoxicology. 2009;30(1):127-35.
Xie HR, Hu LS, Li, GY. SH-SY5Y human neuroblastoma cell line: in vitro cell model of dopaminergic neurons in Parkinson's disease. Chin Med J. 2010;123(8):1086-92.
Gimenez‐Cassina A, Lim F, Diaz‐Nido J. Differentiation of a human neuroblastoma into neuron‐like cells increases their susceptibility to transduction by herpesviral vectors. J Neurosci. Res. 2006;84(4):755-67.
Kovalevich J, Langford D. Considerations for the use of SH-SY5Y neuroblastoma cells in neurobiology. Neuronal Cell Culture: Methods and Protocol: Amini S & White M;2013:9-21
Mosmann T. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J Immunol Methods. 1983;65(1–2):55-63.
Al-Sheddi ES, Farshori NN, Al-Oqail MM, Musarrat J, Al-Khedhairy AA, Siddiqui MA. Cytotoxicity of Nigella sativa seed oil and extract against human lung cancer cell line. Asian Pac J Cancer P. 2014;15(2):983-987.
Karakas D, Ari F, Ulukaya E. The MTT viability assay yields strikingly false-positive viabilities although the cells are killed by some plant extracts. Turkish J Biol. 2017;41(6):919-25.
Oliveira MT, Rego AC, Morgadinho MT, Macedo TRA, Oliveira CR. Toxic effects of opioid and stimulant drugs on undifferentiated PC12 cells. Annal N Y Acad Sci. 2002;965(1):487-96.
Jamil MFA, Subki MFM, Lan TM, Majid MIA, Adenan MI. The effect of mitragynine on cAMP formation and mRNA expression of mu-opioid receptors mediated by chronic morphine treatment in SK–N–SH neuroblastoma cell. J Ethnopharmacology. 2013;148(1):135-43.
Babazadeh B, Sadeghnia HR, Kapurchal ES, Parsaee H, Nasri S, Tayarani-Najaran Z. Protective effect of Nigella sativa and thymoquinone on serum/glucose deprivation-induced DNA damage in PC12 cells. Avicenna J Phytomedicine. 2012;2(3):125-32.
Ward AS, Kelly TH, Foltin RW, Fischman MW. Effects of d-amphetamine on task performance and social behavior of humans in a residential laboratory. Exp Clin Psychopharm. 1997;5(2):130-6.
Krasnova IN, Ladenheim B, Jayanthi S, et al. Amphetamine-induced toxicity in dopamine terminals in CD-1 and C57BL/6J mice: complex roles for oxygen-based species and temperature regulation. Neuroscience. 2001;107(2):265-74.
Hamdy NM, Taha RA. Effects of Nigella sativa oil and thymoquinone on oxidative stress and neuropathy in streptozotocin-induced diabetic rats. Pharmacology. 2009;84(3):127-34.
Cadet JL, Krasnova NI, Jayanthi S, Lyles J. Neurotoxicity of substituted amphetamines: molecular and cellular mechanisms. Neurotox Res. 2007;11(34):183-202.
Hotchkiss AJ, Gibb JW. Long-term effects of multiple doses of methamphetamine on tryptophan hydroxylase and tyrosine hydroxylase activity in rat brain. J Pharmacol Exp Ther. 1980; 214(2):257-62.
Richfield EK, Penney JB, Young AB. Anatomical and affinity state comparisons between dopamine D1 and D2 receptors in the rat central nervous system. Neuroscience. 1989;30(3):767-77.
Daberkow DP, Brown HD, Bunner KD. et al. Amphetamine paradoxically augments exocytotic dopamine release and phasic dopamine signals. J Neurosci. 2013;33(2):452-63.
Schmitz Y, Lee CJ, Schmauss C, Gonon F, Sulzer D. Amphetamine distorts stimulation-dependent dopamine overflow: effects on D2 autoreceptors, transporters, and synaptic vesicle stores. J Neuroscience. 2001;21(16):5916-24.
Yamamoto BK, Moszczynska A, Gudelsky GA. Amphetamine toxicities. Annal N Y Acad Sci. 2010;1187(1):101-121.
El-Shamy AK, Khadrawy AY, El-Feki AM, Refaat HI, Sawie GH. The effect of both vitamin E and thymoquinone on monoamine neurotransmitter changes induced by nicotine treatment and withdrawal in the cortex and hippocampus of rat brain. J App Sci Res. 2013;9(6):4030-4040.
Farkhondeh T, Samarghandian S, Shahri AMP, Samini F. The neuroprotective effects of thymoquinone: A review. Dose-Response. 2018;16(2):1-11.
Cunha-Oliveira T, Rego AC, Oliveira CR. Cellular and molecular mechanisms involved in the neurotoxicity of opioid and psychostimulant drugs. Brain Res Rev. 2008;58(1):192-208.
Mansour MA, Nagi MN, El‐Khatib AS, Al‐Bekairi AM. Effects of thymoquinone on antioxidant enzyme activities, lipid peroxidation and DT‐diaphorase in different tissues of mice: a possible mechanism of action. Cell Biochem Func. 2002;20(2):143-51.
Alhebshi AH, Gotoh M, Suzuki I. Thymoquinone protects cultured rat primary neurons against amyloid β-induced neurotoxicity. Biochemical and Biophysical Research Communications. 2013;433(4):362-7.
Badary OA, Taha RA, Gamal El-Din AM, Abdel-Wahab MH. Thymoquinone is a potent superoxide anion scavenger. Drug Chem Toxicol. 2003;26(2):87-98.
Daba MH, Abdel-Rahman MS. Hepatoprotective activity of thymoquinone in isolated rat hepatocytes. Toxicol Lett. 1998;95(1):23-9.
Harzallah HJ, Grayaa R, Kharoubi W, Maaloul A, Hammami M, Mahjoub T. Thymoquinone, the Nigella sativa bioactive compound, prevents circulatory oxidative stress caused by 1, 2-dimethylhydrazine in erythrocyte during colon postinitiation carcinogenesis. Oxid Med Cell Long. 2012;2012:1-6.
Cobourne-Duval MK, Taka E, Mendonca P, Bauer D, Soliman KF. The antioxidant effects of thymoquinone in activated BV-2 murine microglial cells. Neurochem Res. 2016;41(12):3227-38.
Khan A, Vaibhav K, Javed H. Attenuation of Aβ-induced neurotoxicity by thymoquinone via inhibition of mitochondrial dysfunction and oxidative stress. Mol Cell Biochem. 2012; 369(1-2):55-65.
- Abstract Viewed: 386 times
- PDF Downloaded: 193 times