Skip to main content
SpringerLink
Log in

Ac-cel, a Novel Antioxidant, Protects Against Hydrogen Peroxide-induced Injury in PC12 Cells via Attenuation of Mitochondrial Dysfunction

  • Published:
Journal of Molecular Neuroscience Aims and scope Submit manuscript

Abstract

Oxidative stress has been implicated in pathophysiology of many neurodegenerative diseases (ND) and increased oxidative stress is closely associated with mitochondrial dysfunction. As a result, looking for potent antioxidants, especially those targeting mitochondria, has become an attractive strategy in ND therapy. In this study, we explored protective effects and potential mechanism of Ac-cel, a novel compound, against hydrogen peroxide (H2O2)-induced injury in PC12 cells. Pretreatment of PC12 cells with Ac-cel prior to 24 h of H2O2 exposure markedly attenuated cytotoxicity induced by H2O2 as evidenced by morphological changes and 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Ac-cel also exhibited potent antiapoptotic effect demonstrated by results of annexin V and PI staining. The above beneficial effects of Ac-cel were accompanied by improved mitochondrial function, reduced caspase-3 cleavage as well as upregulated ratio of Bcl-2/Bax protein expression. Moreover, Ac-cel pretreatment markedly reversed intracellular reactive oxygen species (ROS) accumulation following 30 min of H2O2 exposure in PC12 cells. Further, subcellular investigation indicated that Ac-cel significantly reduced production of mitochondrial ROS in isolated rat cortical mitochondria. Taken together, the present study, for the first time, reports that Ac-cel pretreatment inhibits H2O2-stimulated early accumulation of intracellular ROS possibly via reducing mitochondrial ROS production directly and leads to subsequent preservation of mitochondrial function. These results indicate that Ac-cel is a potential drug candidate for treatment of oxidative stress-associated ND.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

Abbreviations

CNS:

Central nervous system

DCF-DA:

2, 7-Dichlorofluorescein diacetate

DMEM:

Dulbecco's modified Eagle's medium

DMSO:

Dimethylsulfoxide

H2O2 :

Hydrogen peroxide

MMP:

Mitochondrial membrane potential

MPTP:

Mitochondrial permeability transition pore

MTT:

3-(4,5-Dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide

NCS:

Newborn calf serum

ND:

Neurodegenerative diseases

PC12:

Pheochromocytoma

PI:

Propidium iodide

ROS:

Reactive oxygen species

SD:

Sprague–Dawley

References

  • Abdel-Kader R, Hauptmann S, Keil U et al (2007) Stabilization of mitochondrial function by Ginkgo biloba extract (EGb 761). Pharmacol Res 56(6):493–502

    Article  PubMed  CAS  Google Scholar 

  • Bae YS, Oh H, Rhee SG, Yoo YD (2011) Regulation of reactive oxygen species generation in cell signaling. Mol Cells 32(6):491–509

    Article  PubMed  CAS  Google Scholar 

  • Benedi J, Arroyo R, Romero C, Martin-Aragon S, Villar AM (2004) Antioxidant properties and protective effects of a standardized extract of Hypericum perforatum on hydrogen peroxide-induced oxidative damage in PC12 cells. Life Sci 75(10):1263–1276

    Article  PubMed  CAS  Google Scholar 

  • Budihardjo I, Oliver H, Lutter M, Luo X, Wang XD (1999) Biochemical pathways of caspase activation during apoptosis. Annu Rev Cell Dev Biol 15:269–290

    Article  PubMed  CAS  Google Scholar 

  • Carafoli E, Rossi CS, Lehninger AL (1964) Cation and anion balance during active accumulation of Ca++ and Mg++ by isolated mitochondria. J Biol Chem 239:3055–3061

    PubMed  CAS  Google Scholar 

  • Chong ZZ, Li F, Maiese K (2005) Oxidative stress in the brain: novel cellular targets that govern survival during neurodegenerative disease. Prog Neurobiol 75(3):207–246

    Article  PubMed  CAS  Google Scholar 

  • Crispo JA, Piche M, Ansell DR et al (2010) Protective effects of methyl gallate on H2O2-induced apoptosis in PC12 cells. Biochem Biophys Res Commun 393(4):773–778

    Article  PubMed  CAS  Google Scholar 

  • Darzynkiewicz Z, Bruno S, Del Bino G et al (1992) Features of apoptotic cells measured by flow cytometry. Cytometry 13(8):795–808

    Article  PubMed  CAS  Google Scholar 

  • Finkel T (2003) Oxidant signals and oxidative stress. Curr Opin Cell Biol 15(2):247–254

    Article  PubMed  CAS  Google Scholar 

  • Gao X, Zheng CY, Yang L, Tang XC, Zhang HY (2009) Huperzine A protects isolated rat brain mitochondria against beta-amyloid peptide. Free Radic Biol Med 46(11):1454–1462

    Article  PubMed  CAS  Google Scholar 

  • Giorgio M, Trinei M, Migliaccio E, Pelicci PG (2007) Hydrogen peroxide: a metabolic by-product or a common mediator of ageing signals? Nat Rev Mol Cell Biol 8(9):722–728

    Article  PubMed  CAS  Google Scholar 

  • Gough DR, Cotter TG (2011) Hydrogen peroxide: a Jekyll and Hyde signalling molecule. Cell Death Dis 2:e213

    Article  PubMed  CAS  Google Scholar 

  • Hakem R, Hakem A, Duncan GS et al (1998) Differential requirement for caspase 9 in apoptotic pathways in vivo. Cell 94(3):339–352

    Article  PubMed  CAS  Google Scholar 

  • Halliwell B (2012) Free radicals and antioxidants: updating a personal view. Nutr Rev 70(5):257–265

    Article  PubMed  Google Scholar 

  • Halliwell B, Aruoma OI (1991) DNA damage by oxygen-derived species. Its mechanism and measurement in mammalian systems. FEBS Lett 281(1–2):9–19

    Article  PubMed  CAS  Google Scholar 

  • Hockenbery DM, Oltvai ZN, Yin XM, Milliman CL, Korsmeyer SJ (1993) Bcl-2 functions in an antioxidant pathway to prevent apoptosis. Cell 75(2):241–251

    Article  PubMed  CAS  Google Scholar 

  • Jomova K, Vondrakova D, Lawson M, Valko M (2010) Metals, oxidative stress and neurodegenerative disorders. Mol Cell Biochem 345(1–2):91–104

    Article  PubMed  CAS  Google Scholar 

  • Jurgensmeier JM, Xie ZH, Deveraux Q, Ellerby L, Bredesen D, Reed JC (1998) Bax directly induces release of cytochrome c from isolated mitochondria. Proc Natl Acad Sci U S A 95(9):4997–5002

    Article  PubMed  CAS  Google Scholar 

  • Kluck RM, BossyWetzel E, Green DR, Newmeyer DD (1997) The release of cytochrome c from mitochondria: a primary site for Bcl-2 regulation of apoptosis. Science 275(5303):1132–1136

    Article  PubMed  CAS  Google Scholar 

  • Koopman G, Reutelingsperger CP, Kuijten GA, Keehnen RM, Pals ST, van Oers MH (1994) Annexin V for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis. Blood 84(5):1415–1420

    PubMed  CAS  Google Scholar 

  • Kroemer G, Zamzami N, Susin SA (1997) Mitochondrial control of apoptosis. Immunol Today 18(1):44–51

    Article  PubMed  CAS  Google Scholar 

  • Niatsetskaya ZV, Sosunov SA, Matsiukevich D et al (2012) The oxygen free radicals originating from mitochondrial complex I contribute to oxidative brain injury following hypoxia–ischemia in neonatal mice. J Neurosci 32(9):3235–3244

    Article  PubMed  CAS  Google Scholar 

  • Ochiai T, Takenaka Y, Kuramoto Y et al (2008) Molecular mechanism for neuro-protective effect of prosaposin against oxidative stress: its regulation of dimeric transcription factor formation. Biochim Biophys Acta 1780(12):1441–1447

    Article  PubMed  CAS  Google Scholar 

  • Pletjushkina OY, Fetisova EK, Lyamzaev KG et al (2005) Long-distance apoptotic killing of cells is mediated by hydrogen peroxide in a mitochondrial ROS-dependent fashion. Cell Death Differ 12(11):1442–1444

    Article  PubMed  CAS  Google Scholar 

  • Scaffidi C, Fulda S, Srinivasan A et al (1998) Two CD95 (APO-1/Fas) signaling pathways. EMBO J 17(6):1675–1687

    Article  PubMed  CAS  Google Scholar 

  • Shen W, Song D, Wu J, Zhang W (2011) Protective effect of a polysaccharide isolated from a cultivated Cordyceps mycelia on hydrogen peroxide-induced oxidative damage in PC12 cells. Phytother Res: PTR 25(5):675–680

    PubMed  CAS  Google Scholar 

  • Shi GX, Liu CZ, Wang LP, Guan LP, Li SQ (2012) Biomarkers of oxidative stress in vascular dementia patients. Can J Neurol Sci 39(1):65–68

    PubMed  Google Scholar 

  • Sims NR, Anderson MF (2008) Isolation of mitochondria from rat brain using Percoll density gradient centrifugation. Nat Protoc 3(7):1228–1239

    Article  PubMed  CAS  Google Scholar 

  • Su MY, Huang HY, Li L, Lu YH (2011) Protective effects of 2′,4′-dihydroxy-6′-methoxy-3′,5′-dimethylchalcone to PC12 cells against cytotoxicity induced by hydrogen peroxide. J Agric Food Chem 59(2):521–527

    Article  PubMed  CAS  Google Scholar 

  • Sultana R, Butterfield DA (2013) Oxidative modification of brain proteins in Alzheimer's disease: perspective on future studies based on results of redox proteomics studies. J Alzheimers Dis 33:S243–51

    PubMed  Google Scholar 

  • Swalwell H, Latimer J, Haywood RM, Birch-Machin MA (2012) Investigating the role of melanin in UVA/UVB- and hydrogen peroxide-induced cellular and mitochondrial ROS production and mitochondrial DNA damage in human melanoma cells. Free Radic Biol Med 52(3):626–634

    Article  PubMed  CAS  Google Scholar 

  • Thannickal VJ, Fanburg BL (2000) Reactive oxygen species in cell signaling. Am J Physiol Lung Cell Mol Physiol 279(6):L1005–1028

    PubMed  CAS  Google Scholar 

  • VanderHeiden MG, Chandel NS, Williamson EK, Schumacker PT, Thompson CB (1997) Bcl-x(L) regulates the membrane potential and volume homeostasis of mitochondria. Cell 91(5):627–637

    Article  CAS  Google Scholar 

  • Wang X, Michaelis EK (2010) Selective neuronal vulnerability to oxidative stress in the brain. Front Aging Neurosci 2:12

    PubMed  Google Scholar 

  • Wang R, Xiao XQ, Tang XC (2001) Huperzine A attenuates hydrogen peroxide-induced apoptosis by regulating expression of apoptosis-related genes in rat PC12 cells. Neuroreport 12(12):2629–2634

    Article  PubMed  CAS  Google Scholar 

  • Xiao XQ, Yang JW, Tang XC (1999) Huperzine A protects rat pheochromocytoma cells against hydrogen peroxide-induced injury. Neurosci Lett 275(2):73–76

    Article  PubMed  CAS  Google Scholar 

  • Zamzami N, Susin SA, Marchetti P et al (1996) Mitochondrial control of nuclear apoptosis. J Exp Med 183(4):1533–1544

    Article  PubMed  CAS  Google Scholar 

  • Zhang HY, Tang XC (2000) Huperzine B, a novel acetylcholinesterase inhibitor, attenuates hydrogen peroxide induced injury in PC12 cells. Neurosci Lett 292(1):41–44

    Article  PubMed  CAS  Google Scholar 

  • Zhang J, Zhu D, Sheng R et al (2009) BZYX, a novel acetylcholinesterase inhibitor, significantly improved chemicals-induced learning and memory impairments on rodents and protected PC12 cells from apoptosis induced by hydrogen peroxide. Eur J Pharmacol 613(1–3):1–9

    Article  PubMed  CAS  Google Scholar 

  • Zhang S, Ye J, Dong G (2010) Neuroprotective effect of baicalein on hydrogen peroxide-mediated oxidative stress and mitochondrial dysfunction in PC12 cells. J Mol Neurosci 40(3):311–320

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

This project is supported by the Ministry of Science and Technology of China (No. 2011CB510004), the National Natural Science Foundation of China (No. 81173034, 81072646), National Science & Technology Major Project “Key New Drug Creation and Manufacturing Program” of China (2012ZX09301001-001, 2012ZX09301001-004), the SKLDR/SIMM Projects (Nos. SIMM1105KF-04 and SIMM1203KF-02), and Shanghai Science and Technology Development Funds (No. 10QA1408100), SA-SIBS Scholarship Program.

Author information

Authors and Affiliations

  1. State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zu Chong Zhi Road, Zhangjiang Hi-Tech Park, Shanghai, 201203, People’s Republic of China

    Xianjun Guo, Yuting Chen, Qunfang Liu, Jian Wu, Luoyi Wang, Xican Tang, Weimin Zhao & Haiyan Zhang

Authors
  1. Xianjun Guo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yuting Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Qunfang Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jian Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Luoyi Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Xican Tang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Weimin Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Haiyan Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Weimin Zhao or Haiyan Zhang.

Additional information

Xianjun Guo and Yuting Chen contribute equally to this work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Guo, X., Chen, Y., Liu, Q. et al. Ac-cel, a Novel Antioxidant, Protects Against Hydrogen Peroxide-induced Injury in PC12 Cells via Attenuation of Mitochondrial Dysfunction. J Mol Neurosci 50, 453–461 (2013). https://doi.org/10.1007/s12031-013-9955-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12031-013-9955-1

Keywords

Search

Navigation