Effects of N-Acetylcysteine on Lead-Exposed PC-12 Cells Article First Online: 20 June 2005 Received: 09 February 2004 Accepted: 13 September 2004 DOI:
Cite this article as: Aykin-Burns, N., Franklin, E.A. & Ercal, N. Arch Environ Contam Toxicol (2005) 49: 119. doi:10.1007/s00244-004-0025-0 Abstract
The neurotoxicity of lead has been well established through numerous studies. However, the cellular processes of lead neurotoxicity, as well as techniques to prevent or reverse cellular damage after lead exposure, remain unknown. If oxidative stress plays a primary role in lead-induced neurotoxicity, antioxidants should assist in reviving lead-exposed cells. The present study explores N-acetylcysteine (NAC) as an antioxidant agent in PC-12 cells after lead exposure. Selective oxidative stress parameters, including glutathione (GSH), glutathione disulfide (GSSG), and malondialdehyde (MDA), were measured in PC-12 cells exposed to various concentrations of lead acetate. Administering NAC after lead exposure improved cell survival as measured by Trypan Blue exclusion. NAC treatment also increased the GSH/GSSG ratio compared to the lead-only group, and reduced MDA to near control levels. These results imply that NAC protects cells from lead-induced oxidative damage by boosting the PC-12 cells’ antioxidant defense mechanisms.
References Antonio, MT, Leret, ML 2000 Study of the neurochemical alterations produced in discrete brain areas by perinatal low-level lead exposure Life Sci 67 635 642 Google Scholar Bradford, MM 1976 A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding Anal Biochem 72 248 254 Google Scholar Bressler, JP, Belloni-Olivi, L, Forman, S, Goldstein, GW 1996 Distinct mechanisms of neurotransmitter release from PC 12 cells exposed to lead J Neurosci Res 46 678 685 Google Scholar Draper, HH, Squires, EJ, Mahmoodi, H, Wu, J, Agarwal, S, Hadley, M 1993 A comparative evaluation of thiobarbituric acid methods for the determination of malondialdehyde in biological materials Free Radic Biol Med 15 353 363 Google Scholar Droge, W, Schulze-Osthoff, K, Mihm, S, Galter, D, Schenk, H, Eck, HP, Roth, S, Gmunder, H 1994 Functions of glutathione and glutathione disulfide in immunology and immunopathology FASEB J 8 1131 1138 Google Scholar Ercal, N, Treeratphan, P, Hammond, TC, Matthews, RH, Grannemann, NH, Spitz, DR 1996a In vivo indices of oxidative stress in lead-exposed C57BL/6 mice are reduced by treatment with meso-2,3-dimercaptosuccinic acid or N-acetylcysteine Free Radic Biol Med 21 157 161 Google Scholar Ercal, N, Treeratphan, P, Lutz, P, Hammond, TC, Matthews, RH 1996b N-acetylcysteine protects Chinese hamster ovary (CHO) cells from lead-induced oxidative stress Toxicology 108 57 64 Google Scholar Goyer, RA 1993 Lead toxicity: current concerns Environ Health Perspect 100 177 187 Google Scholar Gurer, H, Ercal, N 2000 Can antioxidants be beneficial in the treatment of lead poisoning? Free Radic Biol Med 29 927 945 Google Scholar Gurer, H, Ozgunes, H, Oztezcan, S, Ercal, N 1999 Antioxidant role of alpha-lipoic acid in lead toxicity Free Radic Biol Med 27 75 81 Google Scholar Hegg, CC, Miletic, V 1998 Diminished blocking effect of acute lead exposure on high-threshold voltage-gated calcium currents in PC12 cells chronically exposed to the heavy metal Neurotoxicology 19 413 420 Google Scholar Hilliard, A, Ramesh, A, Zawia, NH 1999 Correlation between lead-induced changes in cerebral ornithine decarboxylase and protein kinase C activities during development and in cultured PC 12 cells Int J Dev Neurosci 17 777 785 Google Scholar Hirsch, HV, Mercer, J, Sambaziotis, H, Huber, M, Stark, DT, Torno-Morley, T, Hollocher, K, Ghiradella, H, Ruden, DM 2003 Behavioral effects of chronic exposure to low levels of lead in Drosophila melanogaster Neurotoxicology 24 435 442 Google Scholar Holtzman, D, Olson, JE, DeVries, C, Bensch, K 1987 Lead toxicity in primary cultured cerebral astrocytes and cerebellar granular neurons Toxicol Appl Pharmacol 89 211 225 Google Scholar Hossain, MM, Takashima, A, Nakayama, H, Doi, K 1997 5-Azacytidine induces toxicity in PC12 cells by apoptosis Exp Toxicol Pathol 49 201 206 Google Scholar Kelly, GS 1998 Clinical applications of N-acetylcysteine Altern Med Rev 3 114 127 Google Scholar Kim, K, Annadata, M, Goldstein, GW, Bressler, JP 1997 Induction of c-fos mRNA by lead in PC 12 cells Int J Dev Neurosci 15 175 182 Google Scholar Kuzirian, AM, Epstein, HT, Buck, D, Child, FM, Nelson, T, Alkon, DL 2001 Pavlovian conditioning-specific increases of the Ca2+- and GTP-binding protein, calexcitin in identified Hermissenda visual cells J Neurocytol 30 993 1008 Google Scholar Loikkanen, JJ, Naarala, J, Savolainen, KM 1998 Modification of glutamate-induced oxidative stress by lead: the role of extracellular calcium Free Radic Biol Med 24 377 384 Google Scholar Neal, R, Cooper, K, Kellogg, G, Gurer, H, Ercal, N 1999 Effects of some sulfur-containing antioxidants on lead-exposed lenses Free Radic Biol Med 26 239 243 Google Scholar Needleman, HL, Schell, A, Bellinger, D, Leviton, A, Allred, EN 1990 The long-term effects of exposure to low doses of lead in childhood An 11-year follow-up report. N Engl J Med 322 83 88 Google Scholar Opanashuk, LA, Finkelstein, JN 1995 Relationship of lead-induced proteins to stress response proteins in astroglial cells J Neurosci Res 42 623 632 Google Scholar Oravecz, K, Bazso-Dombi, E, Jeney, F, Nagy, K, Gecse, M, Zs-Nagy, I 2001 The involvement of hydroxyl free radicals in differentiation of the PC-12 rat pheochromocytoma cell line Arch Gerontol Geriatr 33 61 69 Google Scholar Ramesh, GT, Manna, SK, Aggarwal, BB, Jadhav, AL 1999 Lead activates nuclear transcription factor-kappaB, activator protein-1, and amino-terminal c-Jun kinase in pheochromocytoma cells Toxicol Appl Pharmacol 155 280 286 Google Scholar Ridnour, LA, Winters, RA, Ercal, N, Spitz, DR 1999 Measurement of glutathione, glutathione disulfide, and other thiols in mammalian cell and tissue homogenates using high-performance liquid chromatography separation of N-(1-pyrenyl)maleimide derivatives Methods Enzymol 299 258 267 Google Scholar Salinas, JA, Huff, NC 2002 Lead and conditioned fear to contextual and discrete cues Neurotoxicol Teratol 24 541 550 Google Scholar Shelton, KR, Todd, JM, Egle, PM 1986 The induction of stress-related proteins by lead J Biol Chem 261 1935 1940 Google Scholar Suszkiw, JB 2004 Presynaptic disruption of transmitter release by lead Neurotoxicology 25 599 604 Google Scholar Suzukawa, K, Miura, K, Mitsushita, J, Resau, J, Hirose, K, Crystal, R, Kamata, T 2000 Nerve growth factor-induced neuronal differentiation requires generation of Rac1-regulated reactive oxygen species J Biol Chem 275 13175 13178 Google Scholar Wakefield, J 2002 The lead effect? Environ Health Perspect 110 A574 A580 Google Scholar Williams, TM, Ndifor, AM, Near, JT, Reams-Brown, RR 2000 Lead enhances NGF-induced neurite outgrowth in PC12 cells by potentiating ERK/MAPK activation Neurotoxicology 21 1081 1089 Google Scholar Winters, RA, Zukowski, J, Ercal, N, Matthews, RH, Spitz, DR 1995 Analysis of glutathione, glutathione disulfide, cysteine, homocysteine, and other biological thiols by high-performance liquid chromatography following derivatization by n-(1-pyrenyl)maleimide Anal Biochem 227 14 21 Google Scholar Copyright information
© Springer Science+Business Media, Inc. 2005