Skip to main content

Caffeic Acid Phenethyl Ester (CAPE) Protects PC12 Cells from Cisplatin-Induced Neurotoxicity by Activating the NGF-Signaling Pathway

Abstract

Cisplatin is a highly effective chemotherapeutic drug that is toxic to the peripheral nervous system. Findings suggest that axons are early targets of the neurotoxicity of cisplatin. Although many compounds have been reported as neuroprotective, there is no effective treatment against the neurotoxicity of cisplatin. Caffeic acid phenethyl ester (CAPE) is a propolis component with neuroprotective potential mainly attributed to antioxidant and anti-inflammatory mechanisms. We have recently demonstrated the neurotrophic potential of CAPE in a cellular model of neurotoxicity related to Parkinson’s disease. Now, we have assessed the neurotrophic and neuroprotective effects of CAPE against cisplatin-induced neurotoxicity in PC12 cells. CAPE (10 μM) attenuated the inhibition of neuritogenesis and the downregulation of markers of neuroplasticity (GAP-43, synapsin I, synaptophysin, and 200-kD neurofilament) induced by cisplatin (5 μM). This concentration of cisplatin does not affect cell viability, and it was used in order to assess the early neurotoxic events triggered by cisplatin. When a lethal dose of cisplatin was used (IC50 = 32 μM), CAPE (10 μM) increased cell viability. The neurotrophic effect of CAPE is not dependent on NGF nor is it additive to the effect of NGF, but it might involve the activation of the NGF-high-affinity receptors (trkA). The involvement of other neurotrophin receptors such as trkB and trkC is unlikely. This is the first study to demonstrate the protective potential of CAPE against the neurotoxicity of cisplatin and to suggest the involvement of trkA receptors in the neuroprotective mechanism of CAPE. Based on these findings, the beneficial effect of CAPE on cisplatin-induced peripheral neuropathy should be further investigated.

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9

References

  1. Albers JW, Chaudhry V, Cavaletti G , Donehower RC (2014). Interventions for preventing neuropathy caused by cisplatin and related compounds Cochrane Database Syst Rev. CD005228

  2. Avan A, Postma TJ, Ceresa C, Avan A, Cavaletti G, Giovannetti E, Peters GJ (2015) Platinum-induced neurotoxicity and preventive strategies: past, present, and future. Oncologist 20(4):411–432. https://doi.org/10.1634/theoncologist.2014-0044

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  3. Bak J, Kim HJ, Kim SY, Choi YS (2016) Neuroprotective effect of caffeic acid phenethyl ester in 3-nitropropionic acid-induced striatal neurotoxicity. Korean J Physiol Pharmacol 20(3):279–286. https://doi.org/10.4196/kjpp.2016.20.3.279

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  4. Barros Silva R, Santos NA, Martins NM, Ferreira DA, Barbosa F Jr, Oliveira Souza VC, Kinoshita A, Baffa O, Del-Bel E, Santos AC (2013) Caffeic acid phenethyl ester protects against the dopaminergic neuronal loss induced by 6-hydroxydopamine in rats. Neuroscience 233:86–94. https://doi.org/10.1016/j.neuroscience.2012.12.041

    Article  PubMed  CAS  Google Scholar 

  5. Benowitz LI, Routtenberg A (1997) GAP-43: an intrinsic determinant of neuronal development and plasticity. Trends Neurosci 20(2):84–91. https://doi.org/10.1016/S0166-2236(96)10072-2

    Article  PubMed  CAS  Google Scholar 

  6. Burton AW, Fanciullo GJ, Beasley RD, Fisch MJ (2007) Chronic pain in the cancer survivor: a new frontier. Pain Med 8(2):189–198. https://doi.org/10.1111/j.1526-4637.2006.00220.x

    Article  PubMed  Google Scholar 

  7. Calabrese EJ (2008) Enhancing and regulating neurite outgrowth. Crit Rev Toxicol 38(4):391–418. https://doi.org/10.1080/10408440801981981

    Article  PubMed  CAS  Google Scholar 

  8. Cascinu S, Cordella L, del Ferro E, Fronzoni M, Catalano G (1995) Neuroprotective effect of reduced glutathione on cisplatin-based chemotherapy in advanced gastric cancer: a randomized double-blind placebo-controlled trial. J Clin Oncol 13(1):26–32. https://doi.org/10.1200/JCO.1995.13.1.26

    Article  PubMed  CAS  Google Scholar 

  9. Cascinu S, Catalano V, Cordella L, Labianca R, Giordani P, Baldelli AM, Beretta GD, Ubiali E, Catalano G (2002) Neuroprotective effect of reduced glutathione on oxaliplatin-based chemotherapy in advanced colorectal cancer: a randomized, double-blind, placebo-controlled trial. J Clin Oncol 20(16):3478–3483. https://doi.org/10.1200/JCO.2002.07.061

    Article  PubMed  CAS  Google Scholar 

  10. Das KP, Freudenrich TM, Mundy WR (2004) Assessment of PC12 cell differentiation and neuritic growth: a comparison of morphological and neurochemical measures. Neurotoxicol Teratol 26(3):397–406. https://doi.org/10.1016/j.ntt.2004.02.006

    Article  PubMed  CAS  Google Scholar 

  11. Dechant G, Barde YA (2002) The neurotrophin receptor p75(NTR): novel functions and implications for diseases of the nervous system. Nat Neurosci 5(11):1131–1136. https://doi.org/10.1038/nn1102-1131

    Article  PubMed  CAS  Google Scholar 

  12. dos Santos NA, Martins NM, Silva RDEB, Ferreira RS, Sisti FM, dos Santos AC (2014) Caffeic acid phenethyl ester (CAPE) protects PC12 cells from MPP+ toxicity by inducing the expression of neuron-typical proteins. Neurotoxicology 45:131–138. https://doi.org/10.1016/j.neuro.2014.09.007

    Article  PubMed  CAS  Google Scholar 

  13. Edsjo A, Hallberg B, Fagerstrom S, Larsson C, Axelson H, Pahlman S (2001) Differences in early and late responses between neurotrophin-stimulated trkA- and trkC-transfected SH-SY5Y neuroblastoma cells. Cell Growth Differ 12(1):39–50

    PubMed  CAS  Google Scholar 

  14. Ferreira RS, dos Santos NA, Martins NM, Fernandes LS, dos Santos AC (2016a) Non-cytotoxic concentration of cisplatin decreases neuroplasticity-related proteins and neurite outgrowth without affecting the expression of NGF in PC12 cells. Neurochem Res 41(11):2993–3003. https://doi.org/10.1007/s11064-016-2019-5

    Article  PubMed  CAS  Google Scholar 

  15. Ferreira RS, dos Santos NA, Martins NM, Fernandes LS, dos Santos AC (2016b) Non-cytotoxic concentration of cisplatin decreases neuroplasticity-related proteins and neurite outgrowth without affecting the expression of NGF in PC12 cells. Neurochem Res

  16. Flaskos J, Fowler MJ, Teurtrie C, Hargreaves AJ (1999) The effects of carbaryl and trichlorphon on differentiating mouse N2a neuroblastoma cells. Toxicol Lett 110(1-2):79–84. https://doi.org/10.1016/S0378-4274(99)00142-3

    Article  PubMed  CAS  Google Scholar 

  17. HAUSHEER FH, SCHILSKY RL, BAIN S, BERGHORN EJ, LIEBERMAN F (2006) Diagnosis, management, and evaluation of chemotherapy-induced peripheral neuropathy. Semin Oncol 33(1):15–49. https://doi.org/10.1053/j.seminoncol.2005.12.010

    Article  PubMed  CAS  Google Scholar 

  18. HUANG EJ, REICHARDT LF (2001) Neurotrophins: roles in neuronal development and function. Annu Rev Neurosci 24(1):677–736. https://doi.org/10.1146/annurev.neuro.24.1.677

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  19. Huang Y, Jin M, Pi R, Zhang J, Chen M, Ouyang Y, Liu A, Chao X, Liu P, Liu J, Ramassamy C, Qin J (2013) Protective effects of caffeic acid and caffeic acid phenethyl ester against acrolein-induced neurotoxicity in HT22 mouse hippocampal cells. Neurosci Lett 535:146–151. https://doi.org/10.1016/j.neulet.2012.12.051

    Article  PubMed  CAS  Google Scholar 

  20. Ilhan A, Koltuksuz U, Ozen S, Uz E, Ciralik H, Akyol O (1999) The effects of caffeic acid phenethyl ester (CAPE) on spinal cord ischemia/reperfusion injury in rabbits. Eur J Cardiothorac Surg 16(4):458–463. https://doi.org/10.1016/S1010-7940(99)00246-8

    Article  PubMed  CAS  Google Scholar 

  21. Ilhan A, Akyol O, Gurel A, Armutcu F, Iraz M, Oztas E (2004) Protective effects of caffeic acid phenethyl ester against experimental allergic encephalomyelitis-induced oxidative stress in rats. Free Radic Biol Med 37(3):386–394. https://doi.org/10.1016/j.freeradbiomed.2004.04.022

    Article  PubMed  CAS  Google Scholar 

  22. Kaplan DR, Matsumoto K, Lucarelli E, Thiele CJ (1993) Induction of TrkB by retinoic acid mediates biologic responsiveness to BDNF and differentiation of human neuroblastoma cells. Eukaryotic Signal Transduction Group. Neuron 11(2):321–331

    Article  PubMed  CAS  Google Scholar 

  23. Kim HB, Yoo BS (2016) Propolis inhibits neurite outgrowth in differentiating SH-SY5Y human neuroblastoma cells. Toxicol Res 32(3):239–243. https://doi.org/10.5487/TR.2016.32.3.239

    Article  PubMed  PubMed Central  Google Scholar 

  24. Kizilay A, Kalcioglu MT, Ozerol E, Iraz M, Gulec M, Akyol O, Ozturan O (2004) Caffeic acid phenethyl ester ameliorated ototoxicity induced by cisplatin in rats. J Chemother 16(4):381–387. https://doi.org/10.1179/joc.2004.16.4.381

    Article  PubMed  CAS  Google Scholar 

  25. Kurauchi Y, Hisatsune A, Isohama Y, Mishima S, Katsuki H (2012) Caffeic acid phenethyl ester protects nigral dopaminergic neurons via dual mechanisms involving haem oxygenase-1 and brain-derived neurotrophic factor. Br J Pharmacol 166(3):1151–1168. https://doi.org/10.1111/j.1476-5381.2012.01833.x

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  26. L'episcopo F, Serapide MF, Tirolo C, Testa N, Caniglia S, Morale MC, Pluchino S, Marchetti B (2011) A Wnt1 regulated Frizzled-1/beta-Catenin signaling pathway as a candidate regulatory circuit controlling mesencephalic dopaminergic neuron-astrocyte crosstalk: therapeutical relevance for neuron survival and neuroprotection. Mol Neurodegener 6(1):49. https://doi.org/10.1186/1750-1326-6-49

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  27. Longo FM, Massa SM (2013) Small-molecule modulation of neurotrophin receptors: a strategy for the treatment of neurological disease. Nat Rev Drug Discov 12(7):507–525. https://doi.org/10.1038/nrd4024

    Article  PubMed  CAS  Google Scholar 

  28. Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65(1-2):55–63. https://doi.org/10.1016/0022-1759(83)90303-4

    Article  PubMed  CAS  Google Scholar 

  29. Murphy A, Breen KC, Long A, Feighery C, Casey EB, Kelleher D (1993) Neurofilament expression in human T lymphocytes. Immunology 79(1):167–170

    PubMed  PubMed Central  CAS  Google Scholar 

  30. Murtaza G, Karim S, Akram MR, Khan SA, Azhar S, Mumtaz A, Bin Asad MH (2014) Caffeic acid phenethyl ester and therapeutic potentials. Biomed Res Int 2014:145342

    PubMed  PubMed Central  Google Scholar 

  31. Natarajan K, Singh S, Burke TR Jr, Grunberger D, Aggarwal BB (1996) Caffeic acid phenethyl ester is a potent and specific inhibitor of activation of nuclear transcription factor NF-kappa B. Proc Natl Acad Sci U S A 93(17):9090–9095. https://doi.org/10.1073/pnas.93.17.9090

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  32. Ozen S, Akyol O, Iraz M, Sogut S, Ozugurlu F, Ozyurt H, Odaci E, Yildirim Z (2004) Role of caffeic acid phenethyl ester, an active component of propolis, against cisplatin-induced nephrotoxicity in rats. J Appl Toxicol 24(1):27–35. https://doi.org/10.1002/jat.941

    Article  PubMed  CAS  Google Scholar 

  33. Pace A, Savarese A, Picardo M, Maresca V, Pacetti U, del Monte G, Biroccio A, Leonetti C, Jandolo B, Cognetti F, Bove L (2003) Neuroprotective effect of vitamin E supplementation in patients treated with cisplatin chemotherapy. J Clin Oncol 21(5):927–931. https://doi.org/10.1200/JCO.2003.05.139

    Article  PubMed  CAS  Google Scholar 

  34. Phan CW, Lee GS, Hong SL, Wong YT, Brkljaca R, Urban SABD, Malek SN, Sabaratnam V (2014) Hericium erinaceus (Bull.: Fr) Pers. cultivated under tropical conditions: isolation of hericenones and demonstration of NGF-mediated neurite outgrowth in PC12 cells via MEK/ERK and PI3K-Akt signaling pathways. Food Funct 5(12):3160–3169. https://doi.org/10.1039/C4FO00452C

    Article  PubMed  CAS  Google Scholar 

  35. Planting AS, Catimel G, de Mulder PH, de Graeff A, Hoppener F, Verweij J, Oster W, Vermorken JB (1999) Randomized study of a short course of weekly cisplatin with or without amifostine in advanced head and neck cancer. EORTC Head and Neck Cooperative Group. Ann Oncol 10(6):693–700. https://doi.org/10.1023/A:1008353505916

    Article  PubMed  CAS  Google Scholar 

  36. Rasband WS (1997–2014) ImageJ. U. S. National Institutes of Health, Bethesda, Maryland, USA http://imagej.nih.gov/ij/

  37. Sadri S, Bahrami F, Khazaei M, Hashemi M, Asgari A (2010) Cannabinoid receptor agonist WIN-55,212-2 protects differentiated PC12 cells from organophosphorus-induced apoptosis. Int J Toxicol 29(2):201–208. https://doi.org/10.1177/1091581809359708

    Article  PubMed  CAS  Google Scholar 

  38. Santos NAGD, Martins NM, Silva RDB, Ferreira RS, Sisti FM, Santos ACD (2014) Caffeic acid phenethyl ester (CAPE) protects PC12 cells from MPP+ toxicity by inducing the expression of neuron-typical proteins. Neurotoxicology 45:131–138. https://doi.org/10.1016/j.neuro.2014.09.007

    Article  PubMed  CAS  Google Scholar 

  39. Schimmelpfeng J, Weibezahn KF, Dertinger H (2004) Quantification of NGF-dependent neuronal differentiation of PC-12 cells by means of neurofilament-L mRNA expression and neuronal outgrowth. J Neurosci Methods 139(2):299–306. https://doi.org/10.1016/j.jneumeth.2004.05.010

    Article  PubMed  CAS  Google Scholar 

  40. Tapley P, Lamballe F, Barbacid M (1992) K252a is a selective inhibitor of the tyrosine protein kinase activity of the trk family of oncogenes and neurotrophin receptors. Oncogene 7(2):371–381

    PubMed  CAS  Google Scholar 

  41. Theil G (1993) Synapsin I, synapsin II, and synaptophysin: marker proteins for synaptic vesicle. Brain Pathol 3(1):87–95. https://doi.org/10.1111/j.1750-3639.1993.tb00729.x

    Article  Google Scholar 

  42. Tolba MF, Azab SS, Khalifa AE, Abdel-RAHMAN SZ, Abdel-NAIM AB (2013) Caffeic acid phenethyl ester, a promising component of propolis with a plethora of biological activities: a review on its anti-inflammatory, neuroprotective, hepatoprotective, and cardioprotective effects. IUBMB Life 65(8):699–709. https://doi.org/10.1002/iub.1189

    Article  PubMed  CAS  Google Scholar 

  43. Uzar E, Sahin O, Koyuncuoglu HR, Uz E, Bas O, Kilbas S, Yilmaz HR, Yurekli VA, Kucuker H, Songur A (2006) The activity of adenosine deaminase and the level of nitric oxide in spinal cord of methotrexate administered rats: protective effect of caffeic acid phenethyl ester. Toxicology 218(2-3):125–133. https://doi.org/10.1016/j.tox.2005.10.014

    Article  PubMed  CAS  Google Scholar 

  44. van den Berg R, Haenen GRMM, van den Berg H, BAST A (2001) Transcription factor NF-κB as a potential biomarker for oxidative stress. Br J Nutr 86(S1):S121–S127. https://doi.org/10.1079/BJN2001340

    Article  PubMed  Google Scholar 

  45. Vaudry D, Stork PJ, lazarovici P, Eiden LE (2002) Signaling pathways for PC12 cell differentiation: making the right connections. Science 296(5573):1648–1649. https://doi.org/10.1126/science.1071552

    Article  PubMed  CAS  Google Scholar 

  46. Wu G, Fang Y, Lu ZH, Ledeen RW (1998) Induction of axon-like and dendrite-like processes in neuroblastoma cells. J Neurocytol 27:1–14

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

The authors would like to thank CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico, grant number 140106/2015-4, Rafaela Scalco Ferreira) and FAPESP (Fundação de Amparo à Pesquisa do Estado de São Paulo, processo 2017/09332-7) for the financial support.

Author information

Affiliations

Authors

Corresponding author

Correspondence to Antonio Cardozo dos Santos.

Ethics declarations

Conflict of Interest

The authors declare that they have no conflict of interest.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Ferreira, R.S., dos Santos, N.A.G., Martins, N.M. et al. Caffeic Acid Phenethyl Ester (CAPE) Protects PC12 Cells from Cisplatin-Induced Neurotoxicity by Activating the NGF-Signaling Pathway. Neurotox Res 34, 32–46 (2018). https://doi.org/10.1007/s12640-017-9849-z

Download citation

Keywords

  • Cisplatin
  • Peripheral neurotoxicity
  • CAPE
  • Neuroprotection
  • Neuritogenesis
  • trkA receptors