Journal of Natural Medicines

, Volume 66, Issue 3, pp 544–551 | Cite as

Neuroprotective properties of Loranthus parasiticus aqueous fraction against oxidative stress-induced damage in NG108-15 cells

  • Daniel Zin Hua Wong
  • Habsah Abdul KadirEmail author
  • Choy Long Lee
  • Bey Hing Goh
Original Paper


Loranthus parasiticus, a Chinese folk medicine, has been widely used for the treatment of brain diseases, particularly in southwest China. Hence, the present neuroprotection model was designed to investigate its neuroprotective properties against H2O2-induced oxidative stress in NG108-15 cells. L. parasiticus aqueous fraction (LPAF), which was selected in the present study, had proved to be the most active fraction among the other tested extracts and fractions in our previous screening. The restoration of depleted intracellular glutathione (GSH), a major endogenous antioxidant, by LPAF was observed after H2O2 insult. Pretreatment with LPAF substantially reduced the production of intracellular reactive oxygen species generated from H2O2. Apoptotic features such as externalization of phosphatidylserine and disruption of mitochondrial membrane potential were significantly attenuated by LPAF. In addition, cell cycle analysis revealed a prominent decrease in the H2O2-induced sub-G1 population by LPAF. Moreover, apoptotic morphological analysis by DAPI nuclear staining demonstrated that NG108-15 cells treated with H2O2 exhibited apoptotic features, while such changes were greatly reduced in cells pretreated with LPAF. Taken together, these findings confirmed that LPAF exerts marked neuroprotective activity, which raises the possibility of potential therapeutic application of LPAF for managing oxidative stress-related neurological disorders and supports the traditional use of L. parasiticus in treating brain-related diseases.


Apoptosis H2O2 Loranthus parasiticus Neuroprotection Oxidative stress 



We would like to thank University of Malaya for the research funding through Institute of Research and Consultancy (PS238-2008C) and Fundamental Research Grant Scheme (FP009/2010A). We thank Dr. Lee Hong Boon from Cancer Research Initiatives Foundation (CARIF), Sime Darby Medical Centre, Subang Jaya, Selangor, Malaysia for the use of the flow cytometer.


  1. 1.
    Wyllie AH (1980) Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature 284:555–556PubMedCrossRefGoogle Scholar
  2. 2.
    Lo L, Anderson DJ (1995) Postmigratory neural crest cells expressing c-RET display restricted developmental and proliferative capacities. Neuron 15:527–539PubMedCrossRefGoogle Scholar
  3. 3.
    Hara H, Fink K, Endres M, Friedlander RM, Gagliardini V, Yuan JY, Moskowitz MA (1997) Attenuation of transient focal cerebral ischemic injury in transgenic mice expressing a mutant ICE inhibitory protein. J Cereb Blood Flow Metab 17:370–375PubMedCrossRefGoogle Scholar
  4. 4.
    Didier M, Bursztajn S, Adamec E, Passani L, Nixon RA, Coyle JT, Wei JY, Berman SA (1996) DNA strand breaks induced by sustained glutamate excitotoxicity in primary neuronal cultures. J Neurosci 16:2238–2250PubMedGoogle Scholar
  5. 5.
    Palluy O, Rigaud M (1996) Nitric oxide induces cultured cortical neuron apoptosis. Neurosci Lett 208:1–4PubMedCrossRefGoogle Scholar
  6. 6.
    Maiese K (1998) From the bench to the bedside: the molecular management of cerebral ischemia. Clin Neuropharmacol 21:1–7PubMedGoogle Scholar
  7. 7.
    Vincent AM, Maiese K (1999) Nitric oxide induction of neuronal endonuclease activity in programmed cell death. Exp Cell Res 246:290–300PubMedCrossRefGoogle Scholar
  8. 8.
    Bei WJ, Pen WL, Ma Y, Xu AL (2004) NaoXinQing, an anti-stroke herbal medicine, reduces hydrogen peroxide-induced injury in NG108-15 cells. Neurosci Lett 363:262–265PubMedCrossRefGoogle Scholar
  9. 9.
    Kang SS, Lee JY, Choi YK, Kim GS, Han BH (2004) Neuroprotective effects of flavones on hydrogen peroxide-induced apoptosis in SH-SY5Y neuroblostoma cells. Bioorg Med Chem Lett 14:2261–2264PubMedCrossRefGoogle Scholar
  10. 10.
    Yoon Y, Kim KS, Hong SG, Kang BJ, Lee MY, Cho DW (2000) Protective effects of Orostachys japonicus A. Berger (Crassulaceae) on H2O2-induced apoptosis in GT1-1 mouse hypothalamic neuronal cell line. J Ethnopharmacol 69:73–78PubMedCrossRefGoogle Scholar
  11. 11.
    Okuda T, Yoshida T (1987) Corianin from Coriaria japonica A. Gray, and sesquiterpene lactones from Loranthus parasiticus Merr. used for treatment of schizophrenia. Chem Pharm Bull 35:182–187PubMedCrossRefGoogle Scholar
  12. 12.
    Wong DZH, Kadir HA (2011) Antioxidative and neuroprotective effects of Loranthus parasiticus (L.) Merr (Loranthaceae) against oxidative stress in NG108-15 cells. J Med Plant Res 5:6291–6298Google Scholar
  13. 13.
    Griffith OW (1980) Determination of glutathione and glutathione disulfide using glutathione reductase and 2-vinylpyridine. Anal Biochem 106:207–212PubMedCrossRefGoogle Scholar
  14. 14.
    Reers M, Smiley ST, Mottola-Hartshorn C, Chen A, Lin M, Chen LB (1995) Mitochondrial membrane potential monitored by JC-1 dye. Method Enzymol 260:406–417CrossRefGoogle Scholar
  15. 15.
    Dypbukt JM, Ankarcrona M, Burkitt M, Sjoholm A, Strom K, Orrenius S, Nicotera P (1994) Different prooxidant levels stimulate growth, trigger apoptosis, or produce necrosis of insulin-secreting RINm5F cells. The role of intracellular polyamines. J Biol Chem 269:30553–30560PubMedGoogle Scholar
  16. 16.
    Satoh I, Sakai N, Enokido Y, Uchiyama Y, Hatanaka H (1996) Free radical independent protection by nerve growth factor and Bcl-2 of PC12 cells from hydrogen peroxide triggered-apoptosis. J Biochem 120:540–546PubMedCrossRefGoogle Scholar
  17. 17.
    Koh JY, Gwag BJ, Lobner D, Choi DW (1995) Potentiated necrosis of cultured cortical neurons by neurotrophins. Science 268:573–575PubMedCrossRefGoogle Scholar
  18. 18.
    Koh JY, Wie MB, Gwag BJ, Sensi SL, Canzoniero LMT, Demaro J, Csernansky C, Choi DW (1995) Staurosporine-induced neuronal apoptosis. Exp Neurol 135:153–159PubMedCrossRefGoogle Scholar
  19. 19.
    Whittemore ER, Loo DT, Watt JA, Cotman CW (1995) A detailed analysis of hydrogen peroxide-induced cell death in primary neuronal culture. Neuroscience 67:921–932PubMedCrossRefGoogle Scholar
  20. 20.
    Tong L, Perez-Polo JR (1996) Effect of nerve growth factor on AP-1, NF-κB, and Oct DNA binding activity in apoptotic PC12 cells: extrinsic and intrinsic elements. J Neurosci Res 45:1–12PubMedCrossRefGoogle Scholar
  21. 21.
    Maroto R, Perez-Polo JR (1997) BCL-2-related protein expression in apoptosis: Oxidative stress versus serum deprivation in PC12 cells. J Neurochem 69:514–523PubMedCrossRefGoogle Scholar
  22. 22.
    Macho A, Hirsch T, Marzo I, Marchetti P, Dallaporta B, Susin SA, Zamzami N, Kroemer G (1997) Glutathione depletion is an early and calcium elevation is a late event of thymocyte apoptosis. J Immunol 158:4612–4619PubMedGoogle Scholar
  23. 23.
    Halliwell B (1992) Reactive oxygen species and the central nervous system. J Neurochem 59:1609–1623PubMedCrossRefGoogle Scholar
  24. 24.
    Richardson JS, Subbarao KV, Ang LC (1992) On the possible role of iron-induced free radical peroxidation in neural degeneration in Alzheimer’s disease. Ann NY Acad Sci 648:326–327PubMedCrossRefGoogle Scholar
  25. 25.
    Fadok VA, Voelker DR, Campbell PA, Cohen JJ, Bratton DL, Henson PM (1992) Exposure of phosphatidylserine on the surface of apoptotic lymphocytes triggers specific recognition and removal by macrophages. J Immunol 148:2207–2216PubMedGoogle Scholar
  26. 26.
    Koopman G, Reutelingsperger CP, Kuijten GA, Keehnen RM, Pals ST, Van Oers MH (1994) Annexine V for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis. Blood 84:1415–1420PubMedGoogle Scholar
  27. 27.
    Dumont EAWJ, Hofstra L, Van Heerde WL, Eijnde SVD, Doevendans PAF, DeMuinck E, Daemen MARC, Smits JFM, Frederik P, Wellens HJJ, Daemen MJAP, Reutelingsperger CPM (2000) Cardiomyocyte death induced by myocardial ischemia and reperfusion: measurement with recombinant human annexin-V in a mouse model. Circulation 102:1564–1568PubMedGoogle Scholar
  28. 28.
    Zamzami N, Marchetti P, Castedo M, Zanin C, Vayssiere JL, Petit PX, Kroemer G (1995) Reduction in mitochondrial potential constitutes an early irreversible step of programmed lymphocyte death in vivo. J Exp Med 181:1661–1672PubMedCrossRefGoogle Scholar
  29. 29.
    Darzynkiewicz Z, Bruno S, Del Bino G, Gorczyca W, Hotz MA, Lassota P, Traganos F (1992) Features of apoptotic cells measured by flow cytometry. Cytometry 13:795–808PubMedCrossRefGoogle Scholar

Copyright information

© The Japanese Society of Pharmacognosy and Springer 2012

Authors and Affiliations

  • Daniel Zin Hua Wong
    • 1
  • Habsah Abdul Kadir
    • 1
    Email author
  • Choy Long Lee
    • 1
  • Bey Hing Goh
    • 1
  1. 1.Biomolecular Research Group, Biochemistry Program, Institute of Biological Sciences, Faculty of ScienceUniversity of MalayaKuala LumpurMalaysia

Personalised recommendations