Planta Med 2012; 78(17): 1813-1823
DOI: 10.1055/s-0032-1315397
Biological and Pharmacological Activity
Original Papers
Georg Thieme Verlag KG Stuttgart · New York

Antrodia cinnamomea Exhibits a Potent Neuroprotective Effect in the PC12 Cell-Aβ 25–35 Model – Pharmacologically through Adenosine Receptors and Mitochondrial Pathway

Chi-Huang Chang
1   Research Institute of Biotechnology, Hungkuang University, New Taichung City, Taiwan
Hui-Er Wang
2   Institute of Food Science and Applied Technology, Hungkuang University, New Taichung City, Taiwan
Pei-Yu Liaw
1   Research Institute of Biotechnology, Hungkuang University, New Taichung City, Taiwan
Chiung-Chi Peng
3   Graduate Institute of Clinical Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan
Robert Y. Peng
1   Research Institute of Biotechnology, Hungkuang University, New Taichung City, Taiwan
› Author Affiliations
Further Information

Publication History

received 17 April 2012
revised 25 August 2012

accepted 31 August 2012

Publication Date:
11 October 2012 (online)


Antrodia cinnamomea has a diversity of therapeutic effects including anticancer properties. Its neuroprotective effect is rarely cited. We hypothesized that due to its high phenol, triterpenoid, and adenosine contents, it might exhibit a potent neuroprotective effect. The PC12 cell model was used to investigate its pharmaceutical effects. Congo red staining was used to identify the activation of Aβ 25–35. Chemical analysis indicated that the ethanolic extract of Antrodia cinnamomea contained a huge amount (mg/g ethanolic extract of Antrodia cinnamomea) of polyphenolics (133 ± 7), flavonoids (114 ± 6), triterpenoids (175 ± 26), and adenosine (370 ± 17). When tested with Aβ 25–35 (15 µM), the cell viability was suppressed in a dose-dependent fashion with an IC50 value of 10 µM. The biochemical parameters upregulated by Aβ 25–35 (15 µM) involved TNF-α, ROS, MDA, NO, and the intracellular calcium ions. These adverse effects were effectively ameliorated by the ethanolic extract of Antrodia cinnamomea (1 µg/mL). The Western blot analysis revealed that Aβ 25–35 downregulated BcL-2/Bax and upregulated cleaved caspases-9 and − 3 without affecting cleaved caspase-8. The G2/M arrest elicited by Aβ 25–35 was ameliorated by the ethanolic extract of Antrodia cinnamomea. TUNEL assay confirmed the apoptosis, and the ethanolic extract of Antrodia cinnamomea downregulated adenosine A1 and adenosine A2A receptors. Taken together, Aβ 25–35 tends to induce neurotoxicity on PC12 cells. The ethanolic extract of Antrodia cinnamomea is capable of suppressing its neurotoxicity by rescuing the mitochondrial apoptosis pathway and simultaneously by downregulating adenosine A1 and adenosine A2A receptors to retard neurodegeneration and memory dysfunction.

Supporting Information

  • References

  • 1 Kadowaki H, Nishitoh H, Urano F, Sadamitsu C, Matsuzawa A, Takeda K, Masutani H, Yodoi J, Urano Y, Nagano T, Ichijo H. Amyloid beta induces neuronal cell death through ROS-mediated ASK1 activation. Cell Death Differ 2005; 12: 19-24
  • 2 Mattson MP. Pathways towards and away from Alzheimerʼs disease. Nature 2004; 430: 631-639
  • 3 Chang CH, Peng CH, Chen KC, Huang HB, Chiu WT, Peng RY. Shock membrane electropotential drops and limited diffusive distance of β-amyloids in cerebral neurons are detrimental enhancement to Alzheimerʼs diseases. Colloids Surf B Biointerfaces 2009; 73: 339-345
  • 4 Williams TI, Lynn BC, Markesbery WR, Lovell MA. Increased levels of 4-hydroxynonenal and acrolein, neurotoxic markers of lipid peroxidation, in the brain in Mild Cognitive Impairment and early Alzheimerʼs disease. Neurobiol Aging 2006; 27: 1094-1099
  • 5 Lovell MA, Robertson JD, Teesdale WJ, Campell JL, Markesbery WR. Copper, iron and zinc in Alzheimerʼs disease senile plaques. J Neurol Sci 1998; 158: 47-52
  • 6 Butterfield DA, Kanski J. Brain protein oxidation in age-related neurodegenerative disorders that are associated with aggregated proteins. Mech Ageing Dev 2001; 122: 945-962
  • 7 Fowler DM, Koulov AV, Alory-Jost C, Marks MS, Balch WE, Jeffery W, Kelly JW. Functional amyloid formation within mammalian tissue. PLoS Biol 2006; 4: e6
  • 8 Park YH, Kim YJ, Son IH, Yang HD. Inhibition of β-amyloid1-40 peptide aggregation and neurotoxicity by citrate. Korean J Physiol Pharmacol 2009; 13: 273-279
  • 9 Wu SH, Yu ZH, Dai YC, Chen CT, Su CH, Chen LC, Hsu WC, Hwang GY. Taiwanofungus, a polypore new genus. Fungal Sci 2004; 19: 109-116
  • 10 Geethangili M, Tzeng YM. Review of pharmacological effects of Antrodia camphorata and its bioactive compounds. Evid Based Complement Alternat Med 2011; 2011: 212641
  • 11 Chang TT. Biological characteristics and applications of Antrodia cinnamomea . Fungal Sci 2009; 24: 181-185
  • 12 Chung WM, Hui WS, Chu PW, Chiu SW, Ip NY. Ganoderma extract activates MAP kinase and induces the neuronal differentiation of rat pheochromatocytoma PC12 cells. FEBS Lett 2000; 486: 291-296
  • 13 Ao ZH, Xu ZH, Lu ZM, Xu HY, Zhang XM, Dou WF. Niuchangchih (Antrodia camphorata) and its potential in treating liver disease. J Ethnopharmacol 2009; 121: 194-212
  • 14 Peng CC, Chen KC, Peng RY, Su CH, Hsieh-Li HM. Human urinary bladder cancer T24 cells are susceptible to the Antrodia camphorata extracts. Cancer Lett 2006; 243: 109-119
  • 15 Hsieh YC, Rao YK, Wu CC, Huang CY, Geethangili M, Hsu SL, Tzeng YM. Methyl antcinate A from Antrodia camphorata induces apoptosis in human liver cancer cells through oxidant-mediated Cofilin- and Bax-triggered mitochondrial pathway. Chem Res Toxicol 2010; 23: 1256-1267
  • 16 Chang JM, Lee YR, Hung LM, Liu SY, Kuo MT, Wen WC, Chen P. An extract of Antrodia camphorata mycelia attenuates the progression of nephritis in systemic lupus erythematosus-prone NZB/W F1 Mice. Evid Based Complement Alternat Med 2011; 2011: 465894
  • 17 Isakovic AJ, Abbott NJ, Redzic ZB. Brain to blood efflux transport of adenosine: blood-brain barrier studies in the rat. J Neurochem 2004; 90: 272-286
  • 18 Lu MK, Cheng JJ, Lai WL, Lin YR, Huang NK. Adenosine as an active component of Antrodia cinnamomea that prevents rat PC12 cells from serum deprivation-induced apoptosis through the activation of adenosine A(2A) receptors. Life Sci 2006; 79: 252-258
  • 19 Lu MK, Cheng JJ, Lai WL, Lin YR, Huang NK. Fermented Antrodia cinnamomea extract protects rat PC12 cells from serum deprivation-induced apoptosis: the role of the MAPK family. J Agric Food Chem 2008; 56: 865-874
  • 20 Stone TW, Ceruti S, Abbracchio MP. Adenosine receptors and neurological disease: neuroprotection and neurodegeneration. Handb Exp Pharmacol 2009; 193: 535-587
  • 21 Mérot Y, Ferrière F, Debroas E, Flouriot G, Duval D, Saligaut C. Estrogen receptor alpha mediates neuronal differentiation and neuroprotection in PC12 cells: critical role of the Α/Β domain of the receptor. J Mol Endocrinol 2005; 35: 257-267
  • 22 Guo LX, Liu JH, Xia ZN. Geniposide inhibits CoCl2-induced PC12 cells death via the mitochondrial pathway. Chin Med J (Engl) 2009; 122: 2886-2892
  • 23 Poduslo JF, Gilles EJ, Ramakrishnan M, Howell KG, Wengenack TM, Curran GL, Kandimalla KK. HH Domain of Alzheimerʼs disease Aβ provides structural basis for neuronal binding in PC12 and mouse cortical/hippocampal neurons. PLoS One 2010; 5: e8813
  • 24 Taga MS, Miller EE, Pratt DE. Chia seeds as a source of natural lipid antioxidants. J Am Oil Chem Soc 1984; 61: 928-931
  • 25 Hsieh CL, Lin YC, Ko WS, Peng CH, Huang CN, Peng RY. Inhibitory effect of some selected nutraceutic herbs on LDL glycation induced by glucose and glyoxal. J Ethnopharmacol 2005; 102: 357-363
  • 26 Xiang ZB, Tang CH, Chen G, Shi YS. Studies on colorimetric determination of oleanolic acid in Chinese quince. Nat Prod Res Develop 2001; 13: 23-26
  • 27 Klunk WE, Pettergrew JW, Abraham DJ. Quantitative evaluation of congo red binding to amyloid-like proteins with beta-pleated sheet conformation. J Histochem Cytochem 1989; 37: 1273-1281
  • 28 Chang CH, Chen CY, Chiou JY, Peng RY, Peng CH. Astaxanthine secured apoptotic death of PC12 cells induced by β-amyloid peptide 25–35: its molecular action targets. J Med Food 2010; 13: 1-9
  • 29 Nakajima Y, Nishida H, Nakamura Y, Konishi T. Prevention of hydrogen peroxide-induced oxidative stress in PC12 cells by 3,4-dihydroxybenzalacetone isolated from Chaga (Inonotus obliquus (persoon) Pilat). Free Radic Biol Med 2009; 47: 1154-1161
  • 30 Kerry NL, Abbey M. Red wine and fractionated phenolic compounds prepared from red wine inhibit low density lipoprotein oxidation in vitro . Atherosclerosis 1997; 135: 93-102
  • 31 Chen CC, Shiao YJ, Lin RD, Shao YY, Lai MN, Lin CC, Ng LT, Kuo YH. Neuroprotective diterpenes from the fruiting body of A. camphorata . J Nat Prod 2006; 69: 680-691
  • 32 Castillo CA, Albasanz IL, León D, Jordán J, Pallàs M, Camins A, Martín M. Age-related expression of adenosine receptors in brain from the senescence-accelerated mouse. Exp Gerontol 2009; 44: 453-461
  • 33 Fuxe K, Ferré S, Genedani S, Franco R, Agnati LF. Adenosine receptor-dopamine receptor interactions in the basal ganglia and their relevance for brain function. Physiol Behav 2007; 92: 210-217
  • 34 Schiffmann SN, Fisone G, Moresco R, Cunha RA, Ferré S. Adenosine A2A receptors and basal ganglia physiology. Prog Neurobiol 2007; 83: 277-292
  • 35 Cunha RA, Ferré S, Vaugeois JM, Chen JF. Potential therapeutic interest of adenosine A2A receptors in psychiatric disorders. Curr Pharm Des 2008; 14: 1512-1524
  • 36 Ho CM, Huang CC, Huang CJ, Cheng JS, Chen IS, Tsai JY, Jiann BP, Tseng PL, Kuo SJ, Jan CR. Effects of Antrodia camphorata on viability, apoptosis, and [Ca2+]i in PC3 human prostate cancer cells. Chin J Physiol 2008; 51: 78-84 Erratum in: Chin J Physiol 2008; 51: 196 Erratum in: Chin J Physiol 2008; 51: 196
  • 37 Garcia I, Martinou I, Tsujimoto Y, Martinou JC. Prevention of programmed cell death of sympathetic neurons by the bcl-2 proto-oncogene. Science 1992; 258: 302-304
  • 38 Kaeidi A, Esmaeili-Mahani S, Abbasnejad M, Sheibani V, Rasoulian B, Hajializadeh Z, Pasban-Aliabadi H. Satureja khuzestanica attenuates apoptosis in hyperglycemic PC12 cells and spinal cord of diabetic rats. J Nat Med Epub ahead of print March 2, 2012 (PMID: 22391800)
  • 39 Beattie EC, Stellwagen D, Morishita W, Bresnahan JC, Ha BK, von Zastrow M, Beattie MS, Malenka RC. Control of synaptic strength by glial TNF alpha. Science 2002; 295: 2282-2285
  • 40 Stellwagen D, Malenka RC. Synaptic scaling mediated by glial TNF-alpha. Nature 2006; 440: 1054-1059
  • 41 Mark RJ, Lovell MA, Markesbery WR, Uchida K, Mattson MP. A role for 4-hydroxynonenal, an aldehyde product of lipid peroxidation, in disruption of ion homeostasis and neuronal death induced by amyloid beta-peptide. J Neurochem 1997; 68: 255-264
  • 42 Kim BS, Kim MY, Leem YH. Hippocampal neuronal death induced by kainic acid and restraint stress is suppressed by exercise. Neuroscience 2011; 194: 291-301
  • 43 Wang LC, Wang SE, Wang JJ, Tsai TY, Lin CH, Pan TM, Lee CL. In vitro and in vivo comparisons of the effects of the fruiting body and mycelium of Antrodia camphorata against amyloid β-protein-induced neurotoxicity and memory impairment. Appl Microbiol Biotechnol 2012; 94: 1505-1519
  • 44 Maret W. Crosstalk of the group II a and II b metals calcium and zinc in cellular signaling. Proc Natl Acad Sci USA 2001; 98: 12325-12327
  • 45 Bredesen DE, Rao RV, Mehlen P. Cell death in the nervous system. Nature 2006; 443: 796-802
  • 46 Hsu YL, Kuo PL, Cho CY, Ni WC, Tzeng TF, Ng LT, Kuo YH, Lin CC. Antrodia cinnamomea fruiting bodies extract suppresses the invasive potential of human liver cancer cells line PLC/PRF/5 through inhibition of nuclear factor–κB pathway. Food Chem Toxicol 2007; 45: 1249-1257
  • 47 Wang X, Tang X, Li M, Marshall J, Mao Z. Regulation of neuroprotective activity of myocyte-enhancer factor 2 by cAMP-protein kinase A signaling pathway in neuronal survival. J Biol Chem 2005; 280: 16705-16713
  • 48 Pijls NHJ, de Bruyne B. Coronary pressure. Berlin: Springer; 2000. (ISBN 0-7923-6170-9)
  • 49 Kalda A, Yu L, Oztas E, Chen JF. Novel neuroprotection by caffeine and adenosine A(2A) receptor antagonists in animal models of Parkinsonʼs disease. J Neurol Sci 2006; 248: 9-15