Neurotoxicity Research

, Volume 6, Issue 2, pp 119–130 | Cite as

Neuroprotective effect ofH. perforatum extracts on β-amyloid-induced neurotoxicity

  • Bruno A. Silva
  • Alberto C. P. Dias
  • Federico Ferreres
  • João O. Malva
  • Catarina R. Oliveira
Article

Abstract

In the present study we assessed the neuroprotective role of aHypericum perforatum ethanolic extract and obtained fractions in amyloid-β peptide (Aβ)(25–35)-induced cell death in rat cultured hippocampal neurons. Lipid peroxidation was used as a marker of oxidative stress by following the formation of TBARS in rat cortical synaptosomes, after incubation with ascorbate/Fe2+, alone or in the presence of EC97 effective concentrations ofH. perforatum fractions. Induced lipid peroxidation was significantly inhibited by fractions containing flavonol glycosides, flavonol and biflavone aglycones, and by a fraction containing several phenols, mainly chlorogenic acid-type phenolics (21%,77%and 98%, respectively). Lipid peroxidation evaluated after incubation with 25 μM Aβ(25–35), was significantly inhibited byH. perforatum extract.

Cell viability was assessed by use of the Syto-13/PI assay. The total ethanolic extract (TE) and fractions containing flavonol glycosides, flavonol and biflavone aglycones, reduced Aβ(25–35)-induced cell death (65%,58%and 59%,respectively). These results were further supported by morphological analysis of cells stained with cresyl violet. Peptide β-amyloid(25–35) induced a decrease in cell volume, chromatin condensation and nuclear fragmentation, alterations not evident in the presence of the TE and fractions containing hypericins (hypericin concentration = 11.02 μM), or fractions containing flavonoids (quercetin concentration = 21.13 μM). Dendritic lesion,an evidence of neurodegeneration, was observed by neuronal staining with cobalt following insult with Aβ(25–35), but prevented after exposure to the peptide plus the fractions referred above.

The results of the present paper suggest thatH. perforatum extracts may be endowed with neuroprotective compounds able to prevent Aβ(25–35)-induced toxicity.

Keywords

Alzheimer’s disease Hypericum perforatum Amyloid β(25–35) Neuroprotection Hippocampal neurons 

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References

  1. Agostinho P and CR Oliveira (2003) Involvement of calcineurin in the neurotoxic effects induced by amyloid-beta and prion pep-tides.Eur. J. Neurosci. 17, 1189–1196.PubMedCrossRefGoogle Scholar
  2. Ahlemeyer B and J Krieglstein (2003) Neuroprotective effects of Ginkgo biloba extract.Cell. Mol. Life Sci. 60, 1779–1792.PubMedCrossRefGoogle Scholar
  3. Alvarez G, M Ramos, F Ruiz, J Satrustegui and E Bogonez (2003) Pyruvate protection against beta-amyloid-induced neuronal death: role of mitochondrial redox state.J. Neurosci. Res. 73, 260–269.PubMedCrossRefGoogle Scholar
  4. Ambrósio AF, AP Silva, JO Malva, JF Mesquita, AP Carvalho and CM Carvalho (2000) Role of desensitization of AMPA receptors on the neuronal viability and on the [Ca2+]i changes in cultured rat hippocampal neurons.Eur. J. Neurosci. 12, 2021–2031.PubMedCrossRefGoogle Scholar
  5. Areias FM, AC Rego, CR Oliveira and RM Seabra (2001) Antioxidant effect of flavonoids after ascorbate/Fe(2+)-induced oxidative stress in cultured retinal cells.Biochem. Pharmacol. 62, 111–118.PubMedCrossRefGoogle Scholar
  6. Bains JS and CA Shaw (1997) Neurodegenerative disorders in humans: the role of glutathione in oxidative stress-mediated neu-ronal death.Brain Res. Rev. 25, 335–358.PubMedCrossRefGoogle Scholar
  7. Bastianetto S and R Quirion (2002) Natural extracts as possible protective agents of brain aging.Neurobiol. Aging 23, 891–897.PubMedCrossRefGoogle Scholar
  8. Behl C and B Moosmann (2002) Antioxidation neuroprotection in Alzheimer’s disease as preventive and therapeutic approach.Free Radic. Biol. Med. 33, 182–191.PubMedCrossRefGoogle Scholar
  9. Brewer GJ, JR Torricelli, EK Evege and PJ Price (1993) Optimized survival of hippocampal neurons in B27-supplemented Neurobasal, a new serum-free medium combination.J. Neurosci. Res. 25, 567–576.CrossRefGoogle Scholar
  10. Butterweck V (2003) Mechanisms of action of St John’s wort in depression.CNS Drugs 17, 539–562.PubMedCrossRefGoogle Scholar
  11. Cakir A, A Mavi, A Yildirim, ME Duru, M Harmandar and C Kazaz (2003) Isolation and characterization of antioxidant phenolic compounds from the aerial parts of Hypericum hyssopifolium L. by activity-guided fractionation.J. Ethnopharmacol. 87, 73–83.PubMedCrossRefGoogle Scholar
  12. Casley CS, JM Land, MA Sharpe, JB Clark, MR Duchen and L Canevari (2002) Beta-amyloid fragment 25–35 causes mitochon-drial dysfunction in primary cortical neurons.Neurobiol. Dis. 10, 258–267.PubMedCrossRefGoogle Scholar
  13. Conforti F, GA Statti, R Tundis, F Menichini and P Houghton (2002) Antioxidant activity of methanolic extract of Hypericum triquetrifolium Turra aerial part.Fitoterapia 73, 479–483.PubMedCrossRefGoogle Scholar
  14. Cotman CW and JH Su (1996) Mechanisms of neuronal death in Alzheimer’s disease. Brain Pathol.6, 493–506.PubMedCrossRefGoogle Scholar
  15. Di Carlo G, F Borrelli, E Ernst and AA Izzo (2001) St John’s wort: prozac from the plant kingdom.Trends Pharmacol. Sci. 22, 292–297.PubMedCrossRefGoogle Scholar
  16. Dias ACP, RM Seabra, PB Andrade, F Ferreres and MF Ferreira (1999) The development and evaluation of a HPLC-DAD method for the analysis of the phenolic fractions fromin vivo andin vitro biomass of Hypericum species.J. Liquid Chromatography & R.T. 22, 215–217.CrossRefGoogle Scholar
  17. Du HY, M Olivo, BK Tan and BH Bay (2003) Hypericin-mediated photodynamic therapy induces lipid peroxidation and necrosis in nasopharyngeal cancer.Int. J. Oncol. 23, 1401–1405.PubMedGoogle Scholar
  18. Erdelmeier CAJ, E Koch and R Hoerr (2000)Hypericum perfora-tum — St. John’s wort chemical, pharmacological and clinical aspects, In: Atta-ur-Rahman (Ed.)Studies in Natural Products Chemistry — Bioactive Natural Products (Part C) (Elsevier Direct: New York), Vol. 22, pp. 643–716.CrossRefGoogle Scholar
  19. Greeson JM, B Sanford and DA Monti St, (2001) St. John’s wort (Hypericum perforatum): a review of the current pharmacological, toxicological, and clinical literature.Psychopharmacology 153, 402–414.PubMedCrossRefGoogle Scholar
  20. Hensley K, JM Carney, MP Mattson, M Aksenova, M Harris, JF Wu, RA Floyd and DA Butterfield (1994) A model for ß-amyloid aggregation and neurotoxicity based on free radical generation by the peptide: relevance to Alzheimer disease.Proc. Natl. Acad. Sci. USA 94, 3270–3274.CrossRefGoogle Scholar
  21. Hunt EJ, CE Lester, EA Lester and RL Tackett (2002) Effect of St. John’s wort on free radical production.Life Sci. 69, 181–190.CrossRefGoogle Scholar
  22. Jellinger KA (2001) Cell death mechanisms in neurodegeneration.J. Cell. Mol. Med. 5, 1–17.PubMedCrossRefGoogle Scholar
  23. Malva JO, AP Vieira and CR Oliveira (2003) Understanding physiology of glutamate receptors by use of a protocol for neuronal staining.Adv. Physiol. Edu. 27, 78–85.CrossRefGoogle Scholar
  24. Mattson MP (1997) Central role of oxyradicals in the mechanism of amyloid ß-peptide cytotoxicity.Alzheimer Dis. Rev. 2, 1–14.Google Scholar
  25. Mattson MP, J Partin and JG Begley (1998) Amyloid ß-peptide induced apoptosis-related events in synapses and dendrites.Brain Res. 807, 167–176.PubMedCrossRefGoogle Scholar
  26. Melo JB, P Agostinho and CR Oliveira (2002) Amyloid ß-peptide 25-35 reduces [3H]acetylcholine release in retinal neurons. Involvement of metabolic dysfunction.Amyloid 9, 221–228.PubMedGoogle Scholar
  27. Mook-Jung I, H Kim, W Fan, Y Tezuka, S Kadota, H Nishijo and MW Jung (2002) Neuroprotective effects of constituents of the oriental crude drugs, Rhodiola sacra, R. sachalimensis and Tokaku-joki-to against ß-amyloid toxicity, oxidative stress and apoptosis.Biochem. Pharmacol. Bull. 25, 1101–1104.CrossRefGoogle Scholar
  28. Moreira PI, MS Santos, A Moreno and CR Oliveira (2001) Amyloid ß-peptide promotes permeability transition pore in brain mitochondria.Biosci. Rep. 21, 789–800.PubMedCrossRefGoogle Scholar
  29. Moreira PI, MS Santos, AM Moreno and R Seica and CR Oliveira (2003) Increased vulnerability of brain mitochondria in diabetic (Goto-Kakizaki) rats with aging and amyloid-ß exposure.Diabetes 52, 1449–1556.PubMedCrossRefGoogle Scholar
  30. Morimoto K and T Oda (2003) Kainate exacerbates ß-amyloid tox-icity in rat hippocampus.Neurosci. Lett. 340, 242–244.PubMedCrossRefGoogle Scholar
  31. Muller WE, A Singer, M Wonnemann, U Hafner, M Rolliand and C Schafer (1998) Hyperforin represents the neurotransmitter reup-take inhibiting constituent of hypericum extract.Pharmacopsychiatry 31 (Suppl 1): S16-S21.CrossRefGoogle Scholar
  32. Nahrstedt A and V Butterweck (1997) Biologically active and other chemical constituents of the herb ofHypericum perforatum L.Pharmacopshychiatry 30, 129–134.CrossRefGoogle Scholar
  33. Pereira C, MS Santos and CR Oliveira (1999) Involvement of oxidative stress on the impairment of energy metabolism induced by A beta peptides on PC12 cells: protection by antioxidants.Neurobiol. Dis. 6, 209–219.PubMedCrossRefGoogle Scholar
  34. Pereira CF, P Moreira, R Seica, MS Santos and CR Oliveira (2000) Susceptibility to beta-amyloid-induced toxicity is decreased in goto-kakizaki diabetic rats: involvement of oxidative stress.Exp. Neurol. 161, 383–391.PubMedCrossRefGoogle Scholar
  35. Prior RL (2003) Fruits and vegetables in the prevention of cellular oxidative damage.Am. J. Clin. Nutr. 78 (3 Suppl): 570S-578S.PubMedGoogle Scholar
  36. Raina AK, A Hochman, X Zhu, CA Rottkamp, A Nunomura, SL Siedlak, H Boux, RJ Castellani, G Perry and MA Smith (2001) Abortive apoptosis in Alzheimer’s disease.Acta Neuropathol. (Berl.) 101, 305–310.Google Scholar
  37. Rego AC, N Monteiro, AP Silva, JO Malva and RO Catarina (2003) Mitochondrial apoptotic cell death and moderate superoxide generation upon selective activation of non-desensitizing AMPA receptors in hippocampal cultures.J. Neurochem. 86, 792–804.PubMedCrossRefGoogle Scholar
  38. Schempp CM, V Kirkin, B Simon-Haarhaus, A Kersten, J Kiss, CC Termeer, B Gilb, T Kaufmann, C Borner, JP Sleeman and JC Simon (2002) Inhibition of tumour cell growth by hyperforin, a novel anticancer drug from St. John’s wort that acts by induction of apoptosis.Oncogene 21, 1242–1250.PubMedCrossRefGoogle Scholar
  39. Silva AP, AP Carvalho, CM Carvalho and JO Malva (2001) Modulation of intracellular calcium changes and glutamate release by neuropeptide Y1 and Y2 receptors in the rat hippocampus: differential effects in CA1, CA3 and dentate gyrus.J. Neurochem. 79, 286–296.PubMedCrossRefGoogle Scholar
  40. Silva BC and ACP Dias (2002) Evaluation of the free radical scavenger activity ofHypericum perforatum alcoholic extracts.Revista de Fitoterapia 2 (Suppl. 1): p. 133.Google Scholar
  41. Small DH and CA McLean (1999) Alzheimer’s disease and the amyloid ß-protein: what is the role of amyloid?J. Neurochem. 73, 443–449.PubMedCrossRefGoogle Scholar
  42. Tomiyama T, A Shoji, K Kataoka, Y Suwa, S Asano, H Kaneko and N Endo (1996) Inhibition of amyloid ß-protein aggregation and neurotoxicity by rifampicin.J. Biol. Chem. 12, 6839–6844.Google Scholar
  43. Valentäo P, E Fernandes, F Carvalho, PB Andrade, RM Seabra and ML Bastos (2002) Antioxidant activity of Hypericum androsae-mum infusion: scavenging activity against superoxide radical, hydroxyl radical and hypochlorous acid.Biol. Pharm. Bull. 25, 1320–1323.PubMedCrossRefGoogle Scholar
  44. Vila M and S Przedborski (2003) Targeting programmed cell death in neurodegenerative diseases.Nat. Rev. Neurosci. 4, 365–375.PubMedCrossRefGoogle Scholar
  45. Ward MW, AC Rego, BG Frenguelli and DG Nicholls (2000) Mitochondrial membrane potential and glutamate excitotoxicity in cultured cerebellar granule cells.J. Neurosci. 20, 7208–7219.PubMedGoogle Scholar
  46. Yankner B (1996) Mechanisms of neuronal degeneration in Alzheimer’s disease.Neuron 16, 921–932.PubMedCrossRefGoogle Scholar
  47. Youdim KA and JA Joseph (2001) A possible emerging role of phy-tochemicals in improving age-related neurological dysfunctions: a multiplicity of effects.Free Radic. Biol. Med. 30, 583–594.PubMedCrossRefGoogle Scholar
  48. Youdim KA, JP Spencer, H Schroeter and C Rice-Evans (2002) Dietary flavonoids as potential neuroprotectants.Biol. Chem. 383, 503–519.PubMedCrossRefGoogle Scholar

Copyright information

© Springer 2004

Authors and Affiliations

  • Bruno A. Silva
    • 1
    • 2
  • Alberto C. P. Dias
    • 2
  • Federico Ferreres
    • 3
  • João O. Malva
    • 1
  • Catarina R. Oliveira
    • 1
  1. 1.Institute of Biochemistry and Center for Neuroscience and Cell Biology of Coimbra, Faculty of MedicineUniversity of CoimbraCoimbraPortugal
  2. 2.Laboratory of Plant Molecular Biochemistry and Physiology, Department of BiologyUniversity of MinhoBragaPortugal
  3. 3.Research Group on Quality, Safety and Bioactivity of Plant FoodsCEBAS (CSIC)MurciaSpain

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