Abstract
Increased oxidative stress is a widely accepted factor in the development and progression of Alzheimer’s disease. Here, we introduce chitosan, an antioxidant oligosaccharide, as a protective agent against H2O2/FeSO4-induced cell death in the NT2 neural cell line. Chitosan not only protects the neurons against cell death, as measured by MTT and caspase-3 activity, but also decreases amyloid β formation. NT2 neurons can be used to elucidate the relationship between oxidative stress and Aβ formation. We induced Aβ formation through oxidative stress in NT2 neurons and studied the effect of chitosan. We demonstrate that chitosan can be neuroprotective by suppressing Aβ formation. We further show that chitosan exerts its protective effect by up-regulation of HO-1, γ-GCS, Hsp-70, and Nrf2, while it inhibits activation of caspase-3 and NF-κB. Chitosan or chitosan derivatives have potential value as neuroprotective agents, particularly with regard to oxidative stress.
Similar content being viewed by others
Change history
05 February 2022
This article has been retracted. Please see the Retraction Notice for more detail: https://doi.org/10.1007/s11010-022-04373-9
Abbreviations
- AD:
-
Alzheimer’s disease
- AREs:
-
Antioxidant response elements
- DMEM:
-
Dulbecco’s modified Eagle’s medium
- DTT:
-
Dithiothreitol
- DTNB:
-
Dithionitrobenzoic acid
- ECL:
-
Electrochemiluminescence
- ELISA:
-
Sandwich enzyme-linked immunosorbent
- γ-GCS:
-
γ-glutamylcysteine synthetase
- HO-1:
-
Hemeoxigenase-1
- Hsp-70:
-
Heat shock protein-70
- MTT:
-
3-[4, 5-dimethylthiazol-2-yl]-2, 5-dephenyl tetrazolium bromide
- NF-κB:
-
Nuclear factor- κB
- Nrf2:
-
Nuclear factor-erythroid 2 p45-related factor 2
- PBS:
-
Phosphate buffered saline
- PMSF:
-
Phenylmethanesulfonyl fluoride
- tBHQ:
-
tert-butylhydroquinone
References
Gotz ME, Kunig G, Riederer P et al (1994) Oxidative stress: free radical production in neural degeneration. J Pharm Therap 63:37–122
Butterfield DA, Drake J, Pocernich CB et al (2001) Evidence of oxidative damage in Alzheimer’s disease brain: central role for amyloid beta-peptide. Trends Mol Med 7:548–554
Behl C, Davis JB, Lesley R et al (1994) Hydrogen peroxide mediates amyloid beta protein toxicity. J Cell 77:817–827
Chen Q, Liu S, Du Y et al (2006) Carboxymethyl-chitosan protects rabbit chondrocytes frominterleukin-1β-induced apoptosis. Eur J Pharmacol 541:1–8
Jackel RJ, Townsend JA, Kraft AD et al (2007) Nrf2-mediated protection against 6-hydroxydopamine. J Brain Res 1144:192–201
Wurck JC, Goetz EM, Herdegen T et al (2008) Kavalactones protects neural cells against amyloid β peptide-induced neurotoxicity via ERK1/2-dependent Nrf2-activation. J Mol Pharmacol 73:1785–1795
Boothby LA, Doering PL (2005) Vitamin C and Vitamin E for Alzheimer’s disease. Ann Pharmacother 39:2073–2080
Kumar MNV (2000) A review of chitin and chitosan applications. React Funct Polym 46:1–27
Pae HO, Seo WG, Kim NY (2001) Induction of granulocytic differentiation in acute promyelocytic leukemia cells (HL-60) by water-soluble chitosan oligomer. Leuk Res 25:339–346
Yoon HJ, Park HS, Bom HS et al (2005) Chitosan oligosaccharide inhibits 203HgCl2-induced genotoxicity in mice: micronuclei occurrence and chromosomal aberration. Arch Pharm Res 28:1079–1085
Xie W, Xu P, Liu Q (2001) Antioxidant activity of water-soluble chitosan derivatives. Bioorg Med Chem Lett 11:1699–1701
Kamil J, Jeon YJ, Shahidi F (2002) Antioxidative activity of chitosans of different viscosity in cooked comminuted flesh of herring (Clupea harengus). Food Chem 79:69–77
Jeon TI, Hwang SG, Park NG et al (2003) Antioxidative effect of chitosan on chronic carbon tetrachloride induced hepatic injury in rats. Toxicology 187:67–73
Yoon HJ, Moon ME, Park HS et al (2008) Effects of chitosan oligosaccharide (COS) on the glycerol-induced acute renal failure in vitro and in vivo. Food Chem Toxicol 46:710–716
Tokora A, Kobayashi M, Tatekawa N et al (1989) Protective effect of N-acetyl chitose on Listeria monocytogenes infection in mice. Microbiol Immunol 33:357–367
Nishimura K, Nishimura S, Nishi N et al (1984) Immunological activity of chitin and its derivatives. Vaccine 2:93–99
Hirano S (1989) Production and application of chitin and chitosan in Japan. In: Skjak-Braek G, Anthonsen T, Sandford P (eds) Chitin and chitosan. Elsevier Applied Science, London, pp 37–43
Kendra DF, Christian D, Hadwiger LA (1989) Chitosan oligomers from Fusarium solani/pea interactions, chitinase/β-glucanase digestion of sporelings and from fungal wall chitin actively inhibit fungal growth and enhance disease resistance. Physiol Mol Plant Pathol 35:215–230
Uchida Y, Izume M, Ohtakara A (1989) Preparation of chitosan oligomers with purified chitosanase and its application. In: Skjak-Braek G, Anthonsen T, Sandford P (eds) Chitin and chitosan. Elsevier Applied Science, London, pp 373–382
Pleasure SJ, Page C, Lee VMY (1992) Pure, postmitotic, polarized human neurons derived from NTera 2 cells provide a system for expression exogenous proteins in terminally differentiated neurons. J Neurosci 12:1802–1815
Tamango E, Bardini P, Obbili A et al (2002) Oxidative stress increases expression and activity of BACE in NT2 neurons. J Neurobiol Dis 10:279–288
Kutuk O, Basaga H (2003) Aspirin prevents apoptosis and NF-kappaB activation induced by H2O2 in HeLa cells. Free Radic Res 37:1267–1276
Bradford M (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
Ellman GL (1959) Tissue sulfhydryl groups. Arch Biochem Biophys 82:70–77
Miller DK (1997) The role of caspase family of cysteine proteases in apoptosis. Semin Immunol 9:35–49
Keller JN, Kindy MS, Holtsberg FW et al (1998) Mitochondrial MnSOD prevents neural apoptosis and reduces ischemic brain injury: suppression of peroxynitrite production, lipid peroxidaion and mitochondrial dysfunction. J Neurosci 18:687–697
Kruman II, Culmsee C, Chan SL et al (2000) Homocysteine elicits a DNA damage response in neurons that promotes apoptosis and hypersensitivity to excitotoxicity. J Neurosci 20:6920–6926
Frederriske PH, Garland D, Zigler JS et al (1996) Oxidative stress increases production of beta-amyloid precursor protein and beta-amyloid (A-beta) in mammalian lens and A-beta has toxic effects on lens epithelial cells. J Biol Chem 271:10169–10174
Misonou H, Morishima-Kawashima M, Ihara Y (2000) Oxidative stress induces intracellular accumulation of amyloid β-protein (Aβ) in neuroblastoma cells. Biochemistry 39:6951–6959
Paola D, Domenicotti C, Nitti M et al (2000) Oxidative stress induces increase in intracellular amyloid β-protein production and selective activation of βI and βII PKCs in NT2 cells. J Biochem Biophys Res Commun 268:642–646
Tenhunen R, Marver HS, Schmid R (1969) Microsomal heme oxygenase characterization of the enzyme. J Biol Chem 244:6388–6394
Maines MD (1997) The heme oxygenase system: a regulator of second messenger gases. Annu Rev Pharmacol Toxicol 37:517–554
Ponka P (1999) Cell biology of heme. Am J Med Sci 318:241–256
McNally SJ, Harrison EM, Ross JA et al (2007) Curcumin induces heme oxygenase 1 through generation of reactive oxygen species, p38 activation and phosphatase inhibition. Int J Mol Med 19:165–172
Ogborne RM, Rushworth SA, Charalambos CA et al (2004) Heme oxygenase-1: a target for dietary antioxidants. Biochem Soc Trans 32:1003–1005
Hill-Kapturczak N, Thamilselvan V, Liu FY et al (2001) Mechanism of heme oxygenase 1 induction by curcumin in human renal proximal tubule cells. Am J Physiol Renal Physiol 281:F851–F859
Juan SH, Cheng TH, Lin HC et al (2005) Mechanism of concentration dependent induction of heme oxygenase-1 by resveratrol in human aortic smooth muscle cells. Biochem Pharmacol 69:41–48
Alam J, Stewart D, Touchard C et al (1999) Nrf2, a cap’n’collar transcription factor, regulates induction of the heme oxygenase-1gene. J Biol Chem 274:26071–26078
Alam J, Wicks C, Stewart D et al (2000) Mechanism of heme oxygenase-1 gene activation by cadmium in MCF7 mammary epithelial cells. J Biol Chem 275:27694–27702
Balogun E, Hoque M, Gong P et al (2003) Curcumin activates the heam oxygenase-1 gene via regulation of Nrf2 and the antioxidant response element. Biochem J 371:887–895
Rordorf G, Koroshetz WJ, Bonventre JV (1991) Heat shock protects cultured neurons from glutamate toxicity. Neuron 7:1043–1051
Sato K, Saito H, Matsuki N (1996) HSP70 is essential to the neuroprotective effect of heat-shock. Brain Res 740:117–123
Currie RW, Ellison JA, White RF et al (2000) Benign focal ischemic preconditioning induces neuronal Hsp70 and prolonged astrogliosis with expression of Hsp27. Brain Res 863:169–181
Chiu JH, Tsou MT, Tung HH et al (2003) Preconditioned somatothermal stimulation on median nerve territory increases myocardial heat shock protein 70 and protects rat hearts against ischemiareperfusion injury. J Thorac Cardiovasc Surg 125:678–685
Kelly KJ (2005) Heat shock (stress response) proteins and renal ischemia/reperfusion injury. Contrib Nephrol 148:86–106
Kalmar B, Greensmith L (2009) Induction of heat shock proteins for protection against oxidative stress. Adv Drug Deliv Rev 61:310–318
Wild AC, Moinova HR, Mulcahy RT (1999) Regulation of γ-Glutamylcysteine synthetase subunit gene expression by the transcription factor Nrf2. J Biol Chem 274:33627–33636
Lee J, Johnson JA (2004) An important role of Nrf2-ARE pathway in cellular defense mechanism. J Biochem Mol Biol 37:139–143
Sun X, Erb H, Murphy TH (2005) Coordinate regulation of glutathione metabolism in astrocytes by Nrf2. J Biochem Biophys Res Commun 326:371–377
Nguyen T, Sherratt PJ, Pickett CB (2003) Regulatory mechanisms controlling gene expression mediated by the antioxidant response element. Ann Pharmacol Toxicol 43:233–260
Kalayarasan S, Prabhu PN, Sriram N et al (2009) Diallyl sulfide enhances antioxidants and inhibits inflammation through the activation of Nrf2 against gentamicin-induced nephrotoxicity in Wistar rats. Europ J Pharmacol 606:162–171
Longpre F, Garneau P, Christen Y et al (2006) Protection by EGb 761 against β-amyloid induced neurotoxicity: Involvment of NF-κB, SIRT1, and MAPKs pathways and inhibition of amyloid fibril formation. Free Radic Biol Med 41:1781–1794
Wertkin AM, Turner RS, Pleasure SJ et al (1993) Human neurons derived from a teratocarcinoma cell line express solely the 695-amino acid amyloid precursor and produce intracellular beta amyloid or A 4 peptides. J Proc Natl Acad Sci USA 90:9513–9517
Turner RS, Suzuki N, Chuyung AS et al (1996) Amyloid beta 40 and beta 42 are generated intracellulary in cultured human neurons and their secretion increases with maturation. J Biol Chem 271:8966–8970
Tamango E, Aragno M, Parola M et al (2000) NT2 neurons, a classical model for Alzheimer’s disease, are highly susceptible to oxidative stress. Neuroreport 11:1865–1869
Martin D, Rojo AI, Salinas M et al (2004) Regulation of heme oxygenase-1 expression through the phosphatidylinositol 3-kinase/Akt pathway and the Nrf2 transcription factor in response to the antioxidant phytochemical carnosol. J Biol Chem 279:8919–8929
Feinstein DL, Galea E, Reis DJ (1997) Suppression of glial nitric oxide synthase induction by heat shock: effects on proteolytic degradation of IkappaB-alpha. Nitric Oxide 1:167–176
Guzhova IV, Darieva ZA, Melo AR et al (1997) Major stress protein Hsp70 interacts with NF-κB regulatory complex in human T-lymphoma cells. Cell Stress Chaperones 2:132–139
Ran R, Lu A, Zhang L et al (2004) Hsp70 promotes TNF-mediated apoptosis by binding IKK gamma and impairing NF-kappa B survival signaling. Genes Dev 18:1466–1481
Fujikake N, Nagia Y, Popiel HA et al (2008) Heat shock transcription factor 1-activating compounds suppress polyglutamine-induced neurodegeneration through induction of multiple molecular chaperones. JBC 283:26188–26197
Chen X, Dodd G, Thomas S et al (2006) Activation of Nrf2/ARE pathway protects endothelial cells from oxidant injury and inflammatory gene expression. Am J Physiol Heart Circ Physiol 290:1862–1870
Anraku M, Kabashima M, Maruyama T et al (2008) Antioxidant protection of human serum albumin by chitosan. Int J Biol Macromol 43:159–164
Mendis E, Kim MM, Rajapakse N et al (2007) An in vitro cellular analysis of the radical scavenging efficacy of chitooligosaccharide. Life Sci 80:2118–2127
Perskvist N, Long M, Stendahl O et al (2002) Mycobacterium tuberculosis promotes apoptosis in human neutrophils by activating caspase-3 and altering expression of Bax/Bcl-xL via an oxygen-dependent pathway. J Immunol 168:6358–6365
Jiang M, Zhuge X, Yang Y et al (2009) The promotion of peripheral nerve regeneration by chitooligosaccharides in the rat nerve crush injury model. Neurosci Lett 454:239–243
Zhou S, Yang Y, Gu X et al (2008) Chitooligosaccharides protect cultured hippocampal neurons against glutamate-induced neurotoxicity. Neurosci Lett 444:270–274
Acknowledgments
F. KH thanks National elite fund, Iran, for the award of Young Scientist Research Fellowship. This study was supported partially by Shahid Beheshti University (M.C.) Research funds.
Author information
Authors and Affiliations
Corresponding author
About this article
Cite this article
Khodagholi, F., Eftekharzadeh, B., Maghsoudi, N. et al. RETRACTED ARTICLE: Chitosan prevents oxidative stress-induced amyloid β formation and cytotoxicity in NT2 neurons: involvement of transcription factors Nrf2 and NF-κB. Mol Cell Biochem 337, 39–51 (2010). https://doi.org/10.1007/s11010-009-0284-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11010-009-0284-1