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BMC Complementary Medicine and Therapies
Published in: BMC Complementary Medicine and Therapies 1/2013

Open Access 01-12-2013 | Research article

Neurite outgrowth stimulatory effects of culinary-medicinal mushrooms and their toxicity assessment using differentiating Neuro-2a and embryonic fibroblast BALB/3T3

Authors: Chia-Wei Phan, Pamela David, Murali Naidu, Kah-Hui Wong, Vikineswary Sabaratnam

Published in: BMC Complementary Medicine and Therapies | Issue 1/2013

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Abstract

Background

Mushrooms are not only regarded as gourmet cuisine but also as therapeutic agent to promote cognition health. However, little toxicological information is available regarding their safety. Therefore, the aim of this study was to screen selected ethno-pharmacologically important mushrooms for stimulatory effects on neurite outgrowth and to test for any cytotoxicity.

Methods

The stimulatory effect of mushrooms on neurite outgrowth was assessed in differentiating mouse neuroblastoma (N2a) cells. Neurite length was measured using Image-Pro Insight processor system. Neuritogenesis activity was further validated by fluorescence immunocytochemical staining of neurofilaments. In vitro cytotoxicity was investigated by using mouse embryonic fibroblast (BALB/3T3) and N2a cells for any embryo- and neuro-toxic effects; respectively.

Results

Aqueous extracts of Ganoderma lucidum, Lignosus rhinocerotis, Pleurotus giganteus and Grifola frondosa; as well as an ethanol extract of Cordyceps militaris significantly (p < 0.05) promoted the neurite outgrowth in N2a cells by 38.4 ± 4.2%, 38.1 ± 2.6%, 33.4 ± 4.6%, 33.7 ± 1.5%, and 35.8 ± 3.4%; respectively. The IC50 values obtained from tetrazolium (MTT), neutral red uptake (NRU) and lactate dehydrogenase (LDH) release assays showed no toxic effects following 24 h exposure of N2a and 3T3 cells to mushroom extracts.

Conclusion

Our results indicate that G. lucidum, L. rhinocerotis, P. giganteus, G. frondosa and C. militaris may be developed as safe and healthy dietary supplements for brain and cognitive health.
Appendix
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Literature
1.
Bernd P: The role of neurotrophins during early development. Gene Expr. 2008, 14 (4): 241-250. 10.3727/105221608786883799.CrossRefPubMed
2.
Fornasiero EF, Bonanomi D, Benfenati F, Valtorta F: The role of synapsins in neuronal development. Cell Mol Life Sci. 2010, 67: 1383-9136. 10.1007/s00018-009-0227-8.CrossRefPubMed
3.
Wong K-H, Naidu M, David P, Bakar R, Sabaratnam V: Neuroregenerative potential of Lion’s Mane Mushroom, Hericium erinaceus (Bull.: Fr.) Pers. (Higher Basidiomycetes), in the treatment of peripheral nerve injury (Review). Int J Med Mushrooms. 2012, 14 (5): 427-446. 10.1615/IntJMedMushr.v14.i5.10.CrossRefPubMed
4.
Loers G, Schachner M: Recognition molecules and neural repair. J Neurochem. 2007, 101 (4): 865-882. 10.1111/j.1471-4159.2006.04409.x.CrossRefPubMed
5.
Shulman JM, Jager PLD: Evidence for a common pathway linking neurodegenerative diseases. Nat Genet. 2009, 41: 1261-1262. 10.1038/ng1209-1261.CrossRefPubMed
6.
Sabaratnam V, Wong KH, Naidu M, David PR: Neuronal health-Can culinary and medicinal mushrooms help?. J Trad Complem Med. 2011, 3 (1): 62-68.CrossRef
7.
Wong KH, Sabaratnam V, Abdullah N, Naidu M, Keynes R: Activity of aqueous extracts of lions mane mushroom Hericium erinaceus (Bull.: Fr.) Pers. (Aphyllophoromycetideae) on the neural cell line NG108-15. Int J Med Mushrooms. 2007, 9 (1): 57-65. 10.1615/IntJMedMushr.v9.i1.70.CrossRef
8.
Eik LF, Naidu M, David P, Wong KH, Tan YS, Sabaratnam V: Lignosus rhinocerus (Cooke) Ryvarden: A medicinal mushroom that stimulates neurite outgrowth in PC-12 cells. Evid-based Compl Alt. 2012, 2012: 320308-CrossRef
9.
Seow SLS, Naidu M, David P, Wong KH, Sabaratnam V: Potentiation of neuritogenic activity of medicinal mushrooms in rat pheochromocytoma cells. BMC Complem Altern Med. 2013, 13 (1): 157-10.1186/1472-6882-13-157.CrossRef
10.
Phan CW, Wong WL, David P, Naidu M, Sabaratnam V: Pleurotus giganteus (Berk.) Karunarathna & K. D. Hyde: Nutritional value and in vitro neurite outgrowth activity in rat pheochromocytoma cells. BMC Complem Altern Med. 2012, 12: 102-10.1186/1472-6882-12-102.CrossRef
11.
Genschow E, Spielmann H, Scholz G, Pohl I, Seiler A, Clemann N, Bremer S, Becker K: Validation of the embryonic stem cell test in the international ECVAM validation study on three in vitro embryotoxicity tests. Altern Lab Anim. 2004, 32: 209-244.PubMed
12.
Seiler AEM, Spielmann H: The validated embryonic stem cell test to predict embryotoxicity in vitro. Nat Protoc. 2011, 6 (7): 961-978. 10.1038/nprot.2011.348.CrossRefPubMed
13.
Wong WL, Abdulla MA, Chua KH, Rani U, Tan YS, Sabaratnam V: Hepatoprotective effects of Panus giganteus (Berk.) Corner against thioacetamide (TAA)-induced liver injury in rats. Evid-based Compl Alt. 2012, 2012: 170303-
14.
Kanagasabapathy G, Malek SNA, Kuppusamy UR, Vikineswary S: Chemical composition and antioxidant properties of extracts of fresh fruiting bodies of Pleurotus sajor-caju (Fr.) Singer. J Agric Food Chem. 2011, 59 (6): 2618-2626. 10.1021/jf104133g.CrossRefPubMed
15.
Lee ML, Tan NH, Fung SY, Tan CS, Ng ST: The antiproliferative activity of sclerotia of Lignosus rhinocerus (tiger milk mushroom). Evid-based Compl Alt. 2012, 2012: 697603-
16.
Wong KH, Sabaratnam V, Abdullah N, Kuppusamy UR, Naidu M: Effects of cultivation techniques and processing on antimicrobial and antioxidant activities of Hericium erinaceus ( Bull.: Fr.) Pers. extracts. Food Technol Biotechnol. 2009, 47 (1): 47-55.
17.
Wong KH, Naidu M, David P, Abdulla MA, Abdullah N, Kuppusamy UR, Sabaratnam V: Functional recovery enhancement following injury to rodent peroneal nerve by lions mane mushroom, Hericium erinaceus (Bull.: Fr.) Pers. (Aphyllophoromycetideae). Int J Med Mushrooms. 2009, 11 (3): 225-236. 10.1615/IntJMedMushr.v11.i3.20.CrossRef
18.
Wong KH, Naidu M, David P, Abdulla MA, Abdullah N, Kuppusamy UR, Sabaratnam V: Peripheral nerve regeneration following crush injury to rat peroneal nerve by aqueous extract of medicinal mushroom Hericium erinaceus (Bull.: Fr) Pers. (Aphyllophoromycetideae). Evid-based Compl Alt. 2010, 2011: 580752-
19.
Zhou Z, Tang Y, Xiang J, Wua P, Jin H, Wang Z, Morid M, Cai DF: Neuroprotective effects of water-soluble Ganoderma lucidum polysaccharides on cerebral ischemic injury in rats. J Ethnopharmacol. 2010, 131 (1): 154-164. 10.1016/j.jep.2010.06.023.CrossRefPubMed
20.
Lin J, Lin C, Chen M, Ujiie T, Takada A: Radical scavenger and antihepatotoxic activity of Ganoderma formosanum, Ganoderma lucidum and Ganoderma neo-japonicum. J Ethnopharmacol. 1995, 47: 33-41. 10.1016/0378-8741(95)01251-8.CrossRefPubMed
21.
Lee B, Park J, Park J, Shin HJ, Kwon S, Yeom M, Sur B, Kim S, Kim M, Lee H, Yoon SH, Hahm DH: Cordyceps militaris improves neurite outgrowth in Neuro2a cells and reverses memory impairment in rats. Food Sci Biotechnol. 2011, 20 (6): 1599-1608. 10.1007/s10068-011-0221-4.CrossRef
22.
Nishina A, Kimura H, Sekiguchi A, Fukumoto R, Nakajima S, Furukawa S: Lysophosphatidylethanolamine in Grifola frondosa as a neurotrophic activator via activation of MAPK. J Lipid Res. 2006, 47 (7): 1434-1443. 10.1194/jlr.M600045-JLR200.CrossRefPubMed
23.
Flaskos J, Fowler MJ, Teurtrie C, Hargreaves AJ: The effects of carbaryl and trichlorphon on differentiating mouse N2a neuroblastoma cells. Toxicol Lett. 1999, 110: 79-84. 10.1016/S0378-4274(99)00142-3.CrossRefPubMed
24.
Tibuhwa DD: Folk taxonomy and use of mushrooms in communities around Ngorongoro and Serengeti National Park, Tanzania. J Ethnobiol Ethnomed. 2012, 8: 36-10.1186/1746-4269-8-36.CrossRefPubMedPubMedCentral
25.
Cheung WMW, Hui WS, Chu PWK, Chiu SW, Ip NY: Ganoderma extract activates MAP kinases and induces the neuronal differentiation of rat pheochromocytoma PC12 cells. FEBS Lett. 2000, 486 (3): 291-296. 10.1016/S0014-5793(00)02317-6.CrossRefPubMed
26.
27.
Zhang XQ, Ip FCF, Zhang DM, Chen LX, Zhang W, Li YL, Ip NY, Ye WC: Triterpenoids with neurotrophic activity from Ganoderma lucidum. Nat Prod Res. 2011, 25 (17): 1607-1613. 10.1080/14786419.2010.496367.CrossRefPubMed
28.
Zhang R, Xu S, Cai Y, Zhou M, Zuo X, Chan P: Ganoderma lucidum protects dopaminergic neuron degeneration through inhibition of microglial activation. Evid-based Compl Alt. 2011, 2011: 156810-
29.
Lee SS, Chang YS, Noraswati MNR: Utilization of macrofungi by some indigenous communities for food and medicine in Peninsular Malaysia. Forest Ecol Manag. 2009, 257 (10): 2062-2065. 10.1016/j.foreco.2008.09.044.CrossRef
30.
Yu HM, Wang BS, Huang SC, Duh PD: Comparison of protective effects between cultured Cordyceps militaris and natural Cordyceps sinensis against oxidative damage. J Agric Food Chem. 2006, 54: 3132-3138. 10.1021/jf053111w.CrossRefPubMed
31.
Wang X, Wang Z, Yao Y, Li J, Zhang X, Li C, Cheng Y, Ding G, Liu L, Ding Z: Essential role of ERK activation in neurite outgrowth induced by α-lipoic acid. Biochim Biophys Acta. 2011, 1813: 827-838. 10.1016/j.bbamcr.2011.01.027.CrossRefPubMed
32.
Wang Z, Wang J, Li J, Wang X, Yao Y, Zhang X, Cheng Y, Ding G, Liu L: MEK/ERKs signaling is essential for lithium-induced neurite outgrowth in N2a cells. Int J Dev Neurosci. 2011, 29: 415-222. 10.1016/j.ijdevneu.2011.03.001.CrossRefPubMed
33.
Wasilewska-sampaio AP, Silveira MS, Holub O, Goecking R, Gomes CA, Neto VM, Linden R, Ferreira T, Felice FGD: Neuritogenesis and neuronal differentiation promoted by 2,4-dinitrophenol, a novel anti-amyloidogenic compound. FASEB J. 2005, 19: 1627-1636. 10.1096/fj.05-3812com.CrossRefPubMed
34.
Radio NM, Mundy WR: Developmental neurotoxicity testing in vitro: models for assessing chemical effects on neurite outgrowth. Neurotoxicol. 2008, 29 (3): 361-376. 10.1016/j.neuro.2008.02.011.CrossRef
35.
Sofroniew MV, Howe CL, Mobley WC: Nerve growth factor signaling, neuroprotection, and neural repair. Annu Rev Neurosci. 2001, 24: 1217-1281. 10.1146/annurev.neuro.24.1.1217.CrossRefPubMed
36.
Tsuji M, Inanami O, Kuwabara M: Neuroprotective effect of a-phenyl-N-tert-butylnitrone in gerbil hippocampus is mediated by the mitogen-activated protein kinase pathway and heat shock proteins. Neurosci Lett. 2000, 282: 41-44. 10.1016/S0304-3940(00)00844-2.CrossRefPubMed
37.
Zhang W, Liu HT: MAPK signal pathways in the regulation of cell proliferation in mammalian cells. Cell Res. 2002, 12 (1): 9-18. 10.1038/sj.cr.7290105.CrossRefPubMed
38.
Maria AS, Lopez A, Diaz MM, Alba J: Evaluation of the toxicity of Uncaria tomentosa by bioassays in vitro. J Ethnopharmacol. 1997, 57: 183-187. 10.1016/S0378-8741(97)00067-6.CrossRef
39.
Yang F, Lim GP, Begum AN, Ubeda OJ, Simmons MR, Ambegaokar SS, Chen PP, Kayed R, Glabe CG, Frautschy SA, Cole GM: Curcumin inhibits formation of amyloid b oligomers and fibrils, binds plaques, and reduces amyloid in vivo. J Biol Chem. 2005, 280: 5892-58901.CrossRefPubMed
40.
Sasidharan S, Jinxuan O, Latha LY, Amutha S: In vivo toxicity study of Ganoderma boninense. Afr J Pharm Pharmaco. 2011, 5 (16): 1819-1823.
41.
Smina TP, Mathew J, Janardhanan KK, Devasagayam TPA: Antioxidant activity and toxicity profile of total triterpenes isolated from Ganoderma lucidum (Fr.) P. Karst occurring in South India. Environ Toxicol Phar. 2011, 32: 438-446. 10.1016/j.etap.2011.08.011.CrossRef
42.
Chen SN, Nan FH, Chen S, Wu JF, Lu CL, Soni MG: Safety assessment of mushroom β-glucan: Subchronic toxicity in rodents and mutagenicity studies. Food Chem Toxicol. 2011, 49: 2890-2898. 10.1016/j.fct.2011.08.007.CrossRefPubMed
43.
Shien S, Hong N, Yee S, Pailoor J, Mui S: Evaluation of the sub-acute toxicity of the sclerotium of Lignosus rhinocerus (Cooke ), the tiger milk mushroom. J Ethnopharmacol. 2011, 138: 192-200. 10.1016/j.jep.2011.09.004.CrossRef
44.
Weyermann J, Lochmann D, Zimmer A: A practical note on the use of cytotoxicity assays. Int J Pharmaceut. 2005, 288 (2): 369-376. 10.1016/j.ijpharm.2004.09.018.CrossRef
45.
Fotakis G, Timbrell JA: In vitro cytotoxicity assays: Comparison of LDH, neutral red, MTT and protein assay in hepatoma cell lines following exposure to cadmium chloride. Toxicol Lett. 2006, 160: 171-177. 10.1016/j.toxlet.2005.07.001.CrossRefPubMed
Metadata
Title
Neurite outgrowth stimulatory effects of culinary-medicinal mushrooms and their toxicity assessment using differentiating Neuro-2a and embryonic fibroblast BALB/3T3
Authors
Chia-Wei Phan
Pamela David
Murali Naidu
Kah-Hui Wong
Vikineswary Sabaratnam
Publication date
01-12-2013
Publisher
BioMed Central
Published in
BMC Complementary Medicine and Therapies / Issue 1/2013
Electronic ISSN: 2662-7671
DOI
https://doi.org/10.1186/1472-6882-13-261

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