Protective effects of nimodipine and lithium against aluminum-induced cell death and oxidative stress in PC12 cells

Document Type : Original Article


1 Diagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran

2 Faculty of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran


Objective(s): The role of aluminum (Al) in the pathogenesis of neurodegenerative diseases has been implicated in several studies. However, the exact mechanisms of cytotoxic effects of Al have not been elucidated yet. The aim of this study was to investigate the effect of L-type calcium channel antagonist, nimodipine (NM), and lithium chloride (LiCl) on Al-induced toxicity in PC12 cells.
Materials and Methods: PC12 cells were treated with Al-maltolate (Almal) in the presence and absence of different concentrations of NM (50-150 μm) and/or LiCl (0.5-1.0 mm) for 48 hr. Cell viability, apoptosis, and catalase (CAT) activity, a marker of oxidative stress, were then measured using MTT, flow cytometry and enzyme assay, respectively.
Results: The results showed that Almal, dose dependently induced cell death, apoptosis and CAT activity in the PC12 cells. NM significantly increased cell viability and decreased apoptosis and CAT activity of Almal-treated cells in a dose dependent mode. LiCl reduced CAT activity and increased cell viability in Almal-treated cells, without significant effect on apoptosis (P=0.74).
Conclusion: These findings suggest that NM and Li may have benefits in the prevention of Al-induced cytotoxicity through decreasing oxidative stress.


1. Bondy SC. The neurotoxicity of environmental aluminum is still an issue. Neurotoxicology 2010; 31:575-581.
2. Bondy SC, Bondy SC. Low levels of aluminum can lead to behavioral and morphological changes associated with Alzheimer's disease and age-related neurodegeneration. Neurotoxicology 2015; 11.
3. Yuan CY, Lee YJ, Hsu GS. Aluminum overload increases oxidative stress in four functional brain areas of neonatal rats. J Biomed Sci 2012;19:51.
4. Bhalla P, Dhawan DK. Protective role of lithium in ameliorating the aluminium-induced oxidative stress and histological changes in rat brain. Cell Mol Neurobiol 2009; 29:513-521.
5. Kaur A, Gill KD. Disruption of neuronal calcium homeostasis after chronic aluminium toxicity in rats. Basic Clin Pharmacol Toxicol 2005; 96:118-122.
6. Chen L, Liu CJ, Tang M, Li A, Hu XW, Du YM, et al. Action of aluminum on high voltage-dependent calcium current and its modulation by ginkgolide B. Acta Pharmacol Sin 2005; 26:539-545.
7. Chin-Chan M, Navarro-Yepes J, Quintanilla-Vega B. Environmental pollutants as risk factors for neurodegenerative disorders: Alzheimer and Parkinson diseases. Front Cell Neurosci 2015; 9:124.
8. Jinzhu Y, Qinli Z, Jin Y, Pan K, Jianjun H, Qiao N. Aluminum and benzo[a]pyrene co-operate to induce neuronal apoptosis in vitro. J Toxicol Sci 2015; 40: 365-373.
9. Dewitt DA, Hurd JA, Fox N, Townsend BE, Griffioen KJ, Ghribi O, et al. Peri-nuclear clustering of mitochondria is triggered during aluminum maltolate induced apoptosis. J Alzheimers Dis 2006; 9:195-205.
10. Mustafa Rizvi SH, Parveen A, Verma AK, Ahmad I, Arshad M, Mahdi AA. Aluminium induced endoplasmic reticulum stress mediated cell death in SH-SY5Y neuroblastoma cell line is independent of p53. PLoS One 2014; 9:e98409.
11. Allagui MS, Hachani R, Saidi S, Feriani A, Murat JC, Kacem K, et al. Pleiotropic protective roles of melatonin against aluminum-induced toxicity in rats. Gen Physiol Biophys. 2015; 34:415-524.
12. Mohammad NS, Arafa MH, Atteia HH. Coenzyme Q10 and fish oil synergistically alleviate aluminum chloride-induced suppression of testicular steroidogenesis and antioxidant defense. Free Radic Res 2015; 49: 1319-1334.
13. Bhalla P, Garg ML, Dhawan DK. Protective role of lithium during aluminum-induced neurotoxicity. Neurochem Int 2010; 56:256-262.
14. Peters J, Booth A, Peters R. Potential for specific dihydropyridine calcium channel blockers to have a positive impact on cognitive function in humans: a systematic review. Ther Adv chronic Dis 2015; 6: 160-169.
15. Disterhoft JF, Oh MM. Pharmacological and molecular enhancement of learning in aging and Alzheimer's disease. J Physiol Paris 2006 1; 99:180-192.
16. Aslan A, Gurelik M, Cemek M, Buyukokuroglu M, Goksel HM, Eser O. Nimodipine can diminish oxidative stress in patients with severe head trauma. J Neurosurg Sci 2012; 56:247-253.
17. Herzfeld E, Strauss C, Simmermacher S, Bork K, Horstkorte R, Dehghani F, et al. Investigation of the neuroprotective impact of nimodipine on Neuro2a cells by means of a surgery-like stress model. Int J Mol Sci 2014; 15:18453-18465.
18. Shafer TJ, Atchison WD. Transmitter, ion channel and receptor properties of pheochromocytoma (PC12) cells: a model for neurotoxicological studies. Neurotoxicology 1991; 12:473-492.
19. Johnson VJ, Kim SH, Sharma RP. Aluminum-maltolate induces apoptosis and necrosis in neuro-2a cells: potential role for p53 signaling. Toxicol Sci 2005; 83:329-339.
20. Bertholf RL, Herman MM, Savory J, Carpenter RM, Sturgill BC, Katsetos CD, et al. A long-term intravenous model of aluminum maltol toxicity in rabbits: tissue distribution, hepatic, renal, and neuronal cytoskeletal changes associated with systemic exposure. Toxicol Appl Pharmacol 1989; 98:58-74.
21. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 1983; 65:55-63.
22. Saberzadeh J, Arabsolghar R, Takhshid MA. Alpha synuclein protein is involved in Aluminum-induced cell death and oxidative stress in PC12 cells. Brain Res 2016;1635:153-160.
23. Kruger NJ. The Bradford Method for Protein Quantitation. Methods Mol Biol 1994; 32: 9-15.
24. Aebi H. Catalase in vitro. Methods Enzymol 1984; 105: 121-126.
25. Jaishankar M, Tseten T, Anbalagan N, Mathew BB, Beeregowda KN. Toxicity, mechanism and health effects of some heavy metals. Interdiscip Toxicol 2014; 7:60-72.
26. Bork K, Wurm F, Haller H, Strauss C, Scheller C, Gnanapragassam VS, et al. Neuroprotective and neuroregenerative effects of nimodipine in a model system of neuronal differentiation and neurite outgrowth. Molecules 2015; 20:1003-1013.
27. Anekonda TS, Quinn JF, Harris C, Frahler K, Wadsworth TL, Woltjer RL. L-type voltage-gated calcium channel blockade with isradipine as a therapeutic strategy for Alzheimer's disease. Neurobiol Dis 2011; 41:62-70.
28. Archana M, Bastian, Yogesh TL, Kumaraswamy KL. Various methods available for detection of apoptotic cells--a review. Indian J cancer 2013; 50:274-283.
29. John S, Kale M, Rathore N, Bhatnagar D. Protective effect of vitamin E in dimethoate and malathion induced oxidative stress in rat erythrocytes. J Nutr Biochem 2001; 12:500-504.
30. Li ZH, Zlabek V, Velisek J, Grabic R, Machova J, Randak T. Modulation of antioxidant defence system in brain of rainbow trout (Oncorhynchus mykiss) after chronic carbamazepine treatment. Comp Biochem Physiol Toxicol  Pharmacol 2010; 151: 137-141.
31. Kumari K, Khare A, Dange S. The applicability of oxidative stress biomarkers in assessing chromium induced toxicity in the fish Labeo rohita. BioMed Res Int 2014; 2014:782493.
32. Frijhoff J, Winyard PG, Zarkovic N, Davies SS, Stocker R, Cheng D, et al. Clinical Relevance of Biomarkers of Oxidative Stress. Antioxid  Redox Signal 2015; 23:1144-1170.
33. Simms BA, Zamponi GW. Neuronal voltage-gated calcium channels: structure, function, and dysfunction. Neuron 2014; 82:24-45.
34. Sekiya T, Yagihashi A, Asano K, Suzuki S. Nimodipine ameliorates trauma-induced cochlear neuronal death. Neurol Res 2002; 24:775-780.
35. Jia YF, Gao HL, Ma LJ, Li J. Effect of nimodipine on rat spinal cord injury. Genet Mol Res 2015; 14:1269-1276.
36. Li H, Yang X, Shi W, Ma Z, Feng G, Wang Q, et al. Protective effects of nimodipine on cerebrovascular function in chronic alcoholic encephalopathy. Int J Mol Med 2014; 33:201-208.
37. Bailey JM, Hutsell BA, Newland MC. Dietary nimodipine delays the onset of methylmercury neurotoxicity in mice. Neurotoxicology. 2013; 37:108-117.
38. Lecht S, Rotfeld E, Arien-Zakay H, Tabakman R, Matzner H, Yaka R, et al. Neuroprotective effects of nimodipine and nifedipine in the NGF-differentiated PC12 cells exposed to oxygen-glucose deprivation or trophic withdrawal. Int J Dev Neurosci 2012; 30: 465-469.
39. Lockman JA, Geldenhuys WJ, Bohn KA, Desilva SF, Allen DD, Van der Schyf CJ. Differential effect of nimodipine in attenuating iron-induced toxicity in brain- and blood-brain barrier-associated cell types. Neurochem Res 2012; 37:134-142.
40. Kim AH, Sheline CT, Tian M, Higashi T, McMahon RJ, Cousins RJ, et al. L-type Ca(2+) channel-mediated Zn(2+) toxicity and modulation by ZnT-1 in PC12 cells. Brain Res 2000; 886:99-107.
41. Hinkle PM, Osborne ME. Cadmium toxicity in rat pheochromocytoma cells: studies on the mechanism of uptake. Toxicol Appl Pharmacol 1994; 124:91-98.
42. Pachauri V, Mehta A, Mishra D, Flora SJ. Arsenic induced neuronal apoptosis in guinea pigs is Ca2+ dependent and abrogated by chelation therapy: role of voltage gated calcium channels. Neurotoxicology 2013; 35:137-145.
43. Satoh E, Yasuda I, Yamada T, Suzuki Y, Ohyashiki T. Involvement of NO generation in aluminum-induced cell death. Biol Pharm Bull 2007; 30:1390-1394.
44. Satoh E, Okada M, Takadera T, Ohyashiki T. Glutathione depletion promotes aluminum-mediated cell death of PC12 cells. Biol Pharm Bull 2005; 28:941-946.
45. Vota DM, Crisp RL, Nesse AB, Vittori DC. Oxidative stress due to aluminum exposure induces eryptosis which is prevented by erythropoietin. J Cell Biochem 2012; 113:1581-1589.
46. Kandimalla R, Vallamkondu J, Corgiat EB, Gill KD. Understanding Aspects of aluminum exposure in
Alzheimer's disease development. Brain pathol 2015; 23.
47. Liu ZP, Sun XF. Effect of nimodipine on aluminum induced learning and memory impairment in rats. Chin J Pharmacol Toxicol 1999; 191-193.
48. Yuan Y, Guo JZ, Zhou QX. The homeostasis of iron and suppression of HO-1 involved in the protective effects of nimodipine on neurodegeneration induced by aluminum overloading in mice. Eur J Pharmacol 2008; 586:100-105.
49. Gawlik-Kotelnicka O, Mielicki W, Rabe-Jablonska J, Lazarek J, Strzelecki D. Impact of lithium alone or in combination with haloperidol on oxidative stress parameters and cell viability in SH-SY5Y cell culture. Acta Neuropsychiatr 2016; 28:38-44.
50. Ghribi O, Herman MM, Spaulding NK, Savory J. Lithium inhibits aluminum-induced apoptosis in rabbit hippocampus, by preventing cytochrome c translocation, Bcl-2 decrease, Bax elevation and caspase-3 activation. J neurochem 2002; 82:137-145.
51. Beurel E, Grieco SF, Jope RS. Glycogen synthase kinase-3 (GSK3): regulation, actions, and diseases. Pharmacol Ther 2015; 148:114-131.