Cellular and Molecular Neurobiology

, Volume 34, Issue 1, pp 101–111 | Cite as

Neuroprotective Effects of Farnesene Against Hydrogen Peroxide-Induced Neurotoxicity In vitro

  • Hasan TurkezEmail author
  • Piera Sozio
  • Fatime Geyikoglu
  • Abdulgani Tatar
  • Ahmet Hacimuftuoglu
  • Antonio Di Stefano
Original Research


Oxidative stress is highly damaging to cellular macromolecules and is also considered a main cause of the loss and impairment of neurons in several neurodegenerative disorders. Recent reports indicate that farnesene (FNS), an acyclic sesquiterpene, has antioxidant properties. However, little is known about the effects of FNS on oxidative stress-induced neurotoxicity. We used hydrogen peroxide (H2O2) exposure for 6 h to model oxidative stress. Therefore, this experimental design allowed us to explore the neuroprotective potential of different FNS isomers (α-FNS and β-FNS) and their mixture (Mix-FNS) in H2O2-induced toxicity in newborn rat cerebral cortex cell cultures for the first time. For this aim, both MTT and lactate dehydrogenase assays were carried out to evaluate cell viability. Total antioxidant capacity (TAC) and total oxidative stress (TOS) parameters were used to assess oxidative alterations. In addition to determining of 8-hydroxy-2-deoxyguanosine (8-OH-dG) levels in vitro, the comet assay was also performed for measuring the resistance of neuronal DNA to H2O2-induced challenge. Our results showed that survival and TAC levels of the cells decreased, while TOS, 8-OH-dG levels and the mean values of the total scores of cells showing DNA damage (comet assay) increased in the group treated with H2O2 alone. But pretreatment of FNS suppressed the cytotoxicity, genotoxicity and oxidative stress, which were increased by H2O2 in clear type of isomers and applied concentration-dependent manners. The order of antioxidant effectiveness for modulating H2O2-induced oxidative stress-based neurotoxicity and genotoxicity is as β-FNS > Mix-FNS > α-FNS.


Farnesene Neuroprotection H2O2 Cultured neuron DNA damage Oxidative stress 



We are grateful to our laboratory specialists for their help and efforts in experiments done in the medical genetics and pharmacology laboratories and animal housing.

Conflict of interest

The authors declare that there are no conflicts of interest.


  1. Abdel-Wahab BA, Metwally ME (2011) Ginkgo biloba enhances the anticonvulsant and neuroprotective effects of sodium valproate against kainic acid-induced seizures in mice. J Pharmacol Toxicol 6:679–690CrossRefGoogle Scholar
  2. Afoulous S, Ferhout H, Raoelison EG, Valentin A, Moukarzel B, Couderc F, Bouajila J (2013) Chemical composition and anticancer, antiinflammatory, antioxidant and antimalarial activities of leaves essential oil of Cedrelopsis grevei. Food Chem Toxicol 56:352–362PubMedCrossRefGoogle Scholar
  3. Al-Maskri AY, Hanif MA, Al-Maskari MY, Abraham AS, Al-sabahi JN, Al-Mantheri O (2011) Essential oil from Ocimum basilicum (Omani Basil): a desert crop. Nat Prod Commun 6:1487–1490PubMedGoogle Scholar
  4. Ban JY, Jeon SY, Nguyen TT, Bae K, Song KS, Seong YH (2006) Neuroprotective effect of oxyresveratrol from smilacis chinae rhizome on amyloid Beta protein (25–35)-induced neurotoxicity in cultured rat cortical neurons. Biol Pharm Bull 29:2419–2424PubMedCrossRefGoogle Scholar
  5. Cacciatore I, Baldassarre L, Fornasari E, Cornacchia C, Di Stefano A, Sozio P, Cerasa LS, Fontana A, Fulle S, Di Filippo ES, La Rovere RM, Pinnen F (2012) (R)-α-lipoyl-glycyl-l-prolyl-l-glutamyl dimethyl ester codrug as a multifunctional agent with potential neuroprotective activities. Chem Med CHem 7:2021–2029PubMedCrossRefGoogle Scholar
  6. Chehregani A, Mohsenzadeh F, Mirazi N, Hajisadeghian S, Baghali Z (2010) Chemical composition and antibacterial activity of essential oils of Tripleurospermum disciforme in three developmental stages. Pharm Biol 48:1280–1284PubMedCrossRefGoogle Scholar
  7. Chen X, Zhang Q, Cheng Q, Ding F (2009) Protective effect of salidroside against H2O2-induced cell apoptosis in primary culture of rat hippocampal neurons. Mol Cell Biochem 32:85–93CrossRefGoogle Scholar
  8. Droge W (2002) Free radicals in the physiological control of cell function. Physiol Rev 82:47–95PubMedGoogle Scholar
  9. Erel O (2004) A novel automated direct measurement method for total antioxidant capacity using a new generation, more stable ABTS radical cation. Clin Biochem 37:277–285PubMedCrossRefGoogle Scholar
  10. Erel O (2005) A new automated colorimetric method for measuring total oxidant status. Clin Biochem 38:1103–1111PubMedCrossRefGoogle Scholar
  11. Farah R, Khamisy-Farah R, Amit T, Youdim MB, Arraf Z (2013) Lithium’s gene expression profile, relevance to neuroprotection A cDNA microarray study. Cell Mol Neurobiol 33:411–420PubMedCrossRefGoogle Scholar
  12. Hou RC, Huang HM, Tzen JT, Jeng KC (2003) Protective effects of sesamin and sesamolin on hypoxic neuronal and PC12 cells. J Neurosci Res 74:123–133PubMedCrossRefGoogle Scholar
  13. Iizuka Y, Hong S, Kim CY, Yang WI, Lee JE, Seong GJ (2010) Protective mechanism of agmatine pretreatment on RGC-5 cells injured by oxidative stress. Braz J Med Biol Res 43:356–358PubMedCrossRefGoogle Scholar
  14. Inman DM, Lambert WS, Calkins DJ, Horner PJ (2013) α-lipoic acid antioxidant treatment limits glaucoma-related retinal ganglion cell death and dysfunction. PLoS ONE 8:e65389PubMedCentralPubMedCrossRefGoogle Scholar
  15. Isobe C, Abe T, Terayama Y (2010) Levels of reduced and oxidized coenzyme Q-10 and 8-hydroxy-2′-deoxyguanosine in the CSF of patients with Alzheimer’s disease demonstrate that mitochondrial oxidative damage and/or oxidative DNA damage contributes to the neurodegenerative process. J Neurol 257:399–404PubMedCrossRefGoogle Scholar
  16. Jain V, Baitharu I, Barhwal K, Prasad D, Singh SB, Ilavazhagan G (2012) Enriched environment prevents hypobaric hypoxia induced neurodegeneration and is independent of antioxidant signaling. Cell Mol Neurobiol 32:599–611PubMedCrossRefGoogle Scholar
  17. Jiang B, Liu JH, Bao Y, An LJ (2003) Hydrogen peroxide-induced apoptosis in PC12 cells and the protective effect of puerarin. Cell Biol Int 27:1025–1031PubMedCrossRefGoogle Scholar
  18. Kang SM, Cha SH, Ko JY, Kang MC, Kim D, Heo SJ, Kim JS, Heu MS, Kim YT, Jung WK, Jeon YJ (2012) Neuroprotective effects of phlorotannins isolated from a brown alga, Ecklonia cava, against H2O2-induced oxidative stress in murine hippocampal HT22 cells. Environ Toxicol Pharmacol 34:96–105PubMedCrossRefGoogle Scholar
  19. Karpińska A, Gromadzka G (2013) Oxidative stress and natural antioxidant mechanisms: the role in neurodegeneration. From molecular mechanisms to therapeutic strategies. Postepy Hig Med Dosw 67:43–53CrossRefGoogle Scholar
  20. Kikuchi Y, Yasuhara T, Agari T, Kondo A, Kuramoto S, Kameda M, Kadota T, Baba T, Tajiri N, Wang F, Tayra JT, Liang H, Miyoshi Y, Borlongan CV, Date I (2011) Urinary 8-OHdG elevations in a partial lesion rat model of Parkinson’s disease correlate with behavioral symptoms and nigrostriatal dopaminergic depletion. J Cell Physiol 226:1390–1398PubMedCrossRefGoogle Scholar
  21. Kumar KH, Khanum F (2013) Hydroalcoholic extract of Cyperus rotundus ameliorates H(2)O (2)-induced human neuronal cell damage via its anti-oxidative and anti-apoptotic machinery. Cell Mol Neurobiol 33:5–17PubMedCrossRefGoogle Scholar
  22. Lewerenz V, Hanelt S, Nastevska C, El-Bahay C, Rouhrdanz E, Kahl R (2003) Antioxidants protect primary rat hepatocyte cultures against acetaminophen-induced DNA strand breaks but not against acetaminophen induced cytotoxicity. Toxicol 191:179–187CrossRefGoogle Scholar
  23. Liu T, Hu HT, Sun QR (2010) Neuroprotective effects of emodin on primary rat cortical neurons apoptosis induced by hydrogen peroxide. Zhong Yao Cai 33:1116–1119PubMedGoogle Scholar
  24. Lorenzo Y, Costa S, Collins AR, Azqueta A (2013) The comet assay, DNA damage, DNA repair and cytotoxicity: hedgehogs are not always dead. Mutagenesis 28:427–432PubMedCrossRefGoogle Scholar
  25. Ma W, Miao Z, Novotny MV (1999) Induction of estrus in grouped female mice (Mus domesticus) by synthetic analogues of preputial gland constituents. Chem Senses 24:289–293PubMedCrossRefGoogle Scholar
  26. Maynard S, Schurman SH, Harboe C, de Souza-Pinto NC, Bohr VA (2009) Base excision repair of oxidative DNA damage and association with cancer and aging. Carcinogenesis 30:2–10PubMedCrossRefGoogle Scholar
  27. McCarthy JJ, Saith S, Linnertz C, Burke JR, Hulette CM, Welsh-Bohmer KA, Chiba-Falek O (2012) The Alzheimer’s associated 5′ region of the SORL1 gene cis regulates SORL1 transcripts expression. Neurobiol Aging 33:1485–1488PubMedCentralPubMedCrossRefGoogle Scholar
  28. Mehri S, Abnous K, Mousavi SH, Shariaty VM, Hosseinzadeh H (2012) Neuroprotective effect of crocin on acrylamide-induced cytotoxicity in PC12 cells. Cell Mol Neurobiol 32:227–235PubMedCrossRefGoogle Scholar
  29. Moslehi M, Meshkini A, Yazdanparast R (2012) Flavonoid baicalein modulates H2O2-induced mitogen-activated protein kinases activation and cell death in SK-N-MC cells. Cell Mol Neurobiol 32:549–560PubMedCrossRefGoogle Scholar
  30. Nabavi SF, Nabavi SM, Habtemariam S, Moghaddam AH, Sureda A, Mirzaei M (2013) Neuroprotective effects of methyl-3-o-methyl gallate against sodium fluoride-induced oxidative stress in the brain of rats. Cell Mol Neurobiol 33:261–267PubMedCrossRefGoogle Scholar
  31. Obuzor GU, Nwaokolo MI (2010) Composition of essential oil of Nigerian (Niger Delta) grown gardenia jasminoide flower. Int Arch Appl Sci Technol 1:32–36Google Scholar
  32. Olmez I, Ozyurt H (2012) Reactive oxygen species and ischemic cerebrovascular disease. Neurochem Int 60:208PubMedCrossRefGoogle Scholar
  33. Pala S, Gurkan H (2008) The role of free radicals in ethiopathogenesis of diseases. Adv Mol Biol 1:1–9Google Scholar
  34. Panieri E, Gogvadze V, Norberg E, Venkatesh R, Orrenius S, Zhivotovsky B (2013) Reactive oxygen species generated in different compartments induce cell death, survival, or senescence. Free Radic Biol Med 57:176–187PubMedCrossRefGoogle Scholar
  35. Sarikurkcu C, Sabih Ozer M, Cakir A, Eskici M, Mete E (2013) GC/MS evaluation and in vitro antioxidant activity of essential oil and solvent extracts of an endemic plant used as folk remedy in Turkey: Phlomis bourgaei Boiss. Evidence Based Comp Alter Med 2013:293080Google Scholar
  36. Shin HT, Chung SH, Lee JS, Kim SS, Shin HD, Jang MH, Shin MC, Bahn GH, Paik EK, Park JH, Kim CJ (2003) Protective effect of shenqi-wan against H2O2-induced apoptosis in hippocampal neuronal cells. Am J Chin Med 31:675–686PubMedCrossRefGoogle Scholar
  37. Si CL, Shen T, Jiang YY, Wu L, Yu GJ, Ren XD, Xu GH, Hu WC (2013). Antioxidant properties and neuroprotective effects of isocampneoside II on hydrogen peroxide-induced oxidative injury in PC12 cells. Food Chem Toxicol (in press) doi: 10.1016/j.fct.2013.05.051
  38. Silva FM, Marques A, Chaveiro A (2010) Reactive oxygen species: a double-edged sword in reproduction. Open Vet Sci J 4:127–133Google Scholar
  39. Singh NP, McCoy MT, Tice RR, Schneider EL (1988) A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res 175:184–191PubMedCrossRefGoogle Scholar
  40. Sozio P, Cerasa LS, Laserra S, Cacciatore I, Cornacchia C, Di Filippo ES, Fulle S, Fontana A, Di Crescenzo A, Grilli M, Marchi M, Di Stefano A (2013) Memantine-sulfur containing antioxidant conjugates as potential prodrugs to improve the treatment of Alzheimer’s disease. Eur J Pharm Sci 49:187–198PubMedCrossRefGoogle Scholar
  41. Stefanova N, Georgievska B, Eriksson H, Poewe W, Wenning GK (2012) Myeloperoxidase inhibition ameliorates multiple system atrophy-like degeneration in a transgenic mouse model. Neurotox Res 21:393–404PubMedCrossRefGoogle Scholar
  42. Sumathi T, Shobana C, Christinal J, Anusha C (2012) Protective effect of Bacopa monniera on methyl mercury-induced oxidative stress in cerebellum of rats. Cell Mol Neurobiol 32:979–987PubMedCrossRefGoogle Scholar
  43. Suntrup S, Teismann I, Bejer J, Suttrup I, Winkels M, Mehler D, Pantev C, Dziewas R, Warnecke T (2013) Evidence for adaptive cortical changes in swallowing in Parkinson’s disease. Brain 36:726–738CrossRefGoogle Scholar
  44. Togar B (2013) The cytological, biochemical and genetic effects of selected sesquiterpenes on healthy neuron and N2a neuroblastoma cell cultures. PhD’s thesis, Atatürk University, Gradute School of Natural and Applied Sciences, Department of Biology, Erzurum, TurkeyGoogle Scholar
  45. Tomassoni D, Amenta F, Amantini C, Farfariello V, Di Mannelli Cesare L, Nwankwo IE, Marini C, Tayebati SK (2013) Brain activity of thioctic Acid enantiomers: in vitro and in vivo studies in an animal model of cerebrovascular injury. Int J Mol Sci 14:4580–4595PubMedCentralPubMedCrossRefGoogle Scholar
  46. Turrens JF (2003) Mitochondrial formation of reactive oxygen species. J Physiol 552:335–344PubMedCrossRefGoogle Scholar
  47. Vlasova IA, Zakharova IO, Sokolova TV, Avrova NF (2013) Metabolic effects of ganglioside GM1 on PC12 cells at oxidative stress depend on modulation of activity of tyrosine kinase of trk receptor. Zh Evol Biokhim Fiziol 49:15–23PubMedGoogle Scholar
  48. Zhang Q, Huang WD, Lv XY, Yang YM (2012) Puerarin protects differentiated PC12 cells from H2O2-induced apoptosis through the PI3 K/Akt signalling pathway. Cell Biol Int 36:419–426PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Hasan Turkez
    • 1
    Email author
  • Piera Sozio
    • 2
  • Fatime Geyikoglu
    • 3
  • Abdulgani Tatar
    • 4
  • Ahmet Hacimuftuoglu
    • 5
  • Antonio Di Stefano
    • 2
  1. 1.Department of Molecular Biology and Genetics, Faculty of ScienceErzurum Technical UniversityErzurumTurkey
  2. 2.Dipartimento di FarmaciaUniversità “G. D’Annunzio”ChietiItaly
  3. 3.Department of Biology, Faculty of ScienceAtatürk UniversityErzurumTurkey
  4. 4.Department of Medical Genetics, Faculty of MedicineAtatürk UniversityErzurumTurkey
  5. 5.Department of Medical Pharmacology, Faculty of MedicineAtatürk UniversityErzurumTurkey

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